EA048216B1 - POLYSPECIFIC ANTIBODIES CONTAINING ONLY HEAVY CHAINS THAT BIND TO CD22 AND CD3 - Google Patents

Description

Cross-references to related applications

This patent application claims priority to U.S. Provisional Patent Application No. 62/861,708, filed June 14, 2019, the disclosure of which is incorporated herein by reference in its entirety.

List of sequences

The present invention contains a Sequence Listing, which has been submitted electronically in ASCII format and is hereby incorporated by reference in its entirety. The ASCII copy created on July 10, 2020 is named TNO-0017-WO_SL.txt and is 115,348 bytes in size.

Field of technology

The present invention relates to human polyspecific heavy chain-only antibodies (e.g., UniAb™) that bind to CD22 and CD3. The present invention also relates to methods for producing such antibodies, compositions, including pharmaceutical compositions, comprising such antibodies, and their use in the treatment of disorders characterized by CD22 expression.

State of the art

CD22.

CD22, also known as SIGLEC-2 (UniProt P20273), is a cell surface receptor expressed on mature B cells. CD22 contains multiple Ig domains and is a member of the immunoglobulin superfamily. The extracellular domain of CD22 interacts with sialic acid moieties, including those present on the cell surface protein CD45. CD22 is thought to function as an inhibitory receptor for B cell receptor signaling. Along with CD20 and CD19, the B cell-restricted expression of CD22 makes it an attractive target for the therapeutic treatment of B cell malignancies. Monoclonal antibodies specific to CD22 have been described in the literature (e.g., Jabbour, Elias, et al. Monoclonal antibodies in acute lymphoblastic leukemia. Blood 125.26 (2015): 4010–4016) and have been used therapeutically as standard monoclonal antibodies (e.g., epratuzumab) and as antibody-drug conjugates (inotuzumab ozogamicin). In addition, CD22-targeted CAR T cells have been used clinically to treat leukemia (Fry, Terry J., et al. CD22-targeted CAR T cells induce remission in B-ALL that is naive or resistant to CD19-targeted CAR immunotherapy. Nature medicine (2017)).

Antibodies containing only heavy chains.

In a typical IgG antibody, the association of the heavy chain and light chain is mediated in part by a hydrophobic interaction between the constant region of the light chain and the CH1 constant domain of the heavy chain. There are additional residues in the heavy chain framework 2 (FR2) and framework 4 (FR4) regions that also contribute to this hydrophobic interaction between the heavy and light chains.

It is known, however, that the serum of camelids (a subgroup of Tylopoda that includes camels, dromedaries and llamas) contains a major type of antibodies consisting exclusively of paired H chains (heavy-chain-only antibodies or UniAbs™). Camelidae UniAbs™ (Camelus dromedarius, Camelus bactrianus, Lama glama, Lama guanaco, Lama alpaca and Lama vicugna) have a unique structure consisting of one variable domain (VHH), a hinge region and two constant domains (CH2 and CH3) that are highly homologous to the CH2 and CH3 domains of classical antibodies. These UniAbs™ lack the first domain of the constant region (CH1), which is present in the genome but is removed during mRNA processing. The absence of the CH1 domain explains the absence of a light chain in the UniAbs™, since this domain is the attachment site for the light chain constant domain. Such UniAbs™ have naturally evolved to confer antigen-binding specificity and high affinity to three CDRs from conventional antibodies, or fragments thereof (Muyldermans, 2001; J Biotechnol 74:277–302; Revets et al., 2005; Expert Opin Biol Ther 5:111–124). Cartilaginous fish such as sharks have also evolved a special type of immunoglobulin, designated IgNAR, which lacks light polypeptide chains and is composed entirely of heavy chains. IgNAR molecules can be manipulated by molecular engineering to produce a variable domain single heavy chain polypeptide (vNAR) (Nuttall et al. Eur. J. Biochem. 270, 3543–3554 (2003); Nuttall et al. Function and Bioinformatics 55, 187–197 (2004); Dooley et al., Molecular Immunology 40, 25–33 (2003)).

The ability of heavy-chain-only antibodies lacking the light chain to bind antigen was established in the 1960s (Jaton et al. (1968) Biochemistry, 7, 4185–4195). The immunoglobulin heavy chain, physically separated from the light chain, retained 80% of the antigen-binding activity of the tetrameric antibody. Sitia et al. (1990) Cell, 60, 781–790 demonstrated that deletion of the CH1 domain from a rearranged mouse μ gene resulted in the production of a heavy-chain-only antibody lacking the light chain in mammalian cell culture. The antibodies produced retained VH-binding specificity and effector functions.

Heavy chain-only antibodies with high specificity and affinity can

- 1 048216 can be produced against various antigens by immunization (van der Linden, R. H., et al. Biochim. Biophys. Acta. 1431, 37-46 (1999)), and the VHH portion can be easily cloned and expressed in yeast (Frenken, L. G. J., et al. J. Biotechnol. 78, 11-21 (2000)). Their expression levels, solubility, and stability are significantly higher than those of classical F(ab) or Fv fragments (Ghahroudi, M. A. et al. FEBS Lett. 414, 521-526 (1997)).

Mice in which the λ (lambda) light (G) chain locus and/or the λ and κ (kappa) L chain loci have been functionally silenced, and antibodies produced by such mice, are described in U.S. Patent Nos. 7,541,513 and 8,367,888. Recombinant production of heavy chain-only antibodies in mice and rats has been described, for example, in WO2006008548; U.S. Patent Application Publication No. 20100122358; Nguyen et al., 2003, Immunology; 109(1), 93-101; Bruggemann et al., Crit. Rev. Immunol.; 2006, 26(5):377-90; and Zou et al, 2007, J Exp Med; 204(13): 3271–3283. Production of knockout rats by embryonic microinjection of zinc finger nuclease is described in Geurts et al., 2009, Science 325(5939):433. Soluble heavy chain-only antibodies and transgenic rodents containing a heterologous heavy chain locus producing such antibodies are described in U.S. Patents 8,883,150 and 9,365,655. CAR-T assemblies containing single-domain antibodies as the binding (targeting) domain are described, for example, in Iri-Sofla et al., 2011, Experimental Cell Research 317:2630–2641 and Jamnani et al., 2014, Biochim Biophys Acta 1840:378–386.

The essence of the invention

Aspects of the invention relate to heavy chain-only antibodies, including but not limited to UniAb™, with binding affinity to CD22. Further aspects of the invention relate to methods for producing such antibodies, compositions comprising such antibodies, and their use in the treatment of disorders characterized by CD22 expression.

Aspects of the invention include multispecific binding compounds that bind to CD3, comprising: a heavy chain variable region comprising: (a) a CDR1 sequence having two or less substitutions in SEQ ID NO: 85; and/or (b) a CDR2 sequence comprising two or less substitutions in SEQ ID NO: 86; and/or (c) a CDR3 sequence comprising two or less substitutions in SEQ ID NO: 87; and a light chain variable region. In some embodiments, the heavy chain CDR1, CDR2, and CDR3 sequences are present in a human VH framework. In some embodiments, the heavy chain variable region comprises heavy chain CDR1, CDR2, and CDR3 sequences in a human VH framework, wherein each CDR sequence comprises a sequence that is at least 85% identical to any one of SEQ ID NOs: 85-87; and the binding compound also comprises a light chain variable region.

In some embodiments, the multispecific binding compound comprises: a heavy chain variable region comprising: (a) a CDR1 sequence having two or less substitutions in SEQ ID NO: 85; and (b) a CDR2 sequence comprising two or less substitutions in SEQ ID NO: 86; and (c) a CDR3 sequence comprising two or less substitutions in SEQ ID NO: 87; and the binding compound also comprises a light chain variable region.

In some embodiments, the multispecific binding compound comprises: a heavy chain variable region comprising a CDR1 sequence of SEQ ID NO: 85, a CDR2 sequence of SEQ ID NO: 86, and a CDR3 sequence of SEQ ID NO: 87; and the binding compound also comprises a light chain variable region.

In some embodiments, the light chain variable region comprises a CDR1, CDR2, and CDR3 sequence in a human VL framework, wherein each CDR sequence comprises a sequence with 3 or fewer amino acid substitutions relative to the CDR sequence or set of CDR sequences in SEQ ID NO: 92; or wherein the CDR sequences comprise a sequence that is at least 85% identical to a CDR sequence or set of CDR sequences in SEQ ID NO: 92. In some embodiments, the light chain variable region comprises a CDR1 sequence of SEQ ID NO: 88, a CDR2 sequence of SEQ ID NO: 89, and a CDR3 sequence of SEQ ID NO: 90. In some embodiments, the heavy chain variable region comprises an amino acid sequence that has at least 95% identity to SEQ ID NO: 91. In some embodiments, the heavy chain variable region comprises the amino acid sequence set forth in SEQ ID NO: 91. In some embodiments, the light chain variable region comprises an amino acid sequence that has at least 95% identity to SEQ ID NO: 92. In some embodiments, the light chain variable region comprises the amino acid sequence of SEQ ID NO: 92.

Aspects of the invention include multispecific binding compounds comprising a first binding unit having a binding affinity for CD22 and a second binding unit having a binding affinity for CD3, wherein the first binding unit comprises: (a) a CDR1 having two or less substitutions in any of the amino acid sequences of SEQ ID NO: 110; and/or (b) a CDR2 having two or less substitutions in any of the amino acid sequences

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SEQ ID NO: 11-17; and/or (c) a CDR3 having two or less substitutions in any of the amino acid sequences of SEQ ID NO: 18-23. In some embodiments, the CDR1, CDR2, and CDR3 sequences of the first binding unit are present in a human framework. In some embodiments, the first binding unit further comprises a heavy chain constant region sequence in the absence of a CH1 sequence.

In some embodiments, the first binding unit comprises a heavy chain variable region comprising: (a) a CDR1 sequence selected from the group consisting of SEQ ID NO: 1-10; and/or (b) a CDR2 sequence selected from the group consisting of SEQ ID NO: 11-17; and/or (c) a CDR3 sequence selected from the group consisting of SEQ ID NO: 18-23.

In some embodiments, the multispecific binding compound comprises: (a) a CDR1 sequence selected from the group consisting of SEQ ID NO: 1-10; and (b) a CDR2 sequence selected from the group consisting of SEQ ID NO: 11-17; and (c) a CDR3 sequence selected from the group consisting of SEQ ID NO: 18-23.

In some embodiments, the multispecific binding compound comprises: (a) a CDR1 sequence of SEQ ID NO: 1, a CDR2 sequence of SEQ ID NO: 11, and a CDR3 sequence of SEQ ID NO: 18; (b) a CDR1 sequence of SEQ ID NO: 1, a CDR2 sequence of SEQ ID NO: 12, and a CDR3 sequence of SEQ ID NO: 19; or (c) a CDR1 sequence of SEQ ID NO: 1, a CDR2 sequence of SEQ ID NO: 12, and a CDR3 sequence of SEQ ID NO: 20. In some embodiments, the multispecific binding compound comprises a heavy chain variable region having at least 95% sequence identity to any one of SEQ ID NOs: 24-84. In some embodiments, the multispecific binding compound comprises a heavy chain variable region sequence selected from the group consisting of SEQ ID NO: 24-84. In some embodiments, the multispecific binding compound comprises the heavy chain variable region sequence of SEQ ID NO: 24.

Aspects of the invention include multispecific binding compounds comprising a first binding unit having a binding affinity for CD22 and a second binding unit having a binding affinity for CD3, wherein the first binding unit comprises a heavy chain variable region comprising: (a) a CDR1 sequence of the formula: G X1 SIX ₂ X ₃ X ₄ X ₅ X ₆ Y (SEQ ID NO: 104), wherein X1 is D or G; X ₂ is S, T, I or N; X ₃ is S or D; X4 is G, S or N; X ₅ is D, G or S; and X ₆ is Y or H; and (b) a CDR2 sequence of the formula: X ₇ X ₈ YX ₉ GX ₁₀ Xu (SEQ ID NO: 105), wherein X ₇ is I or V; X ₈ is Y or H; X ₉ is S or T; X ₁₀ is A, V or S; and Xu is T or A; and (c) a CDR3 sequence of the formula: X ₁₂ RX ₁₃ DSSX ₁₄ WRS (SEQ ID NO: 106), wherein X ₁₂ is T, A or K; X ₁₃ is D or E; and X ₁₄ is N or S.

Aspects of the invention include multispecific binding compounds comprising a first binding unit having a binding affinity for CD22 and a second binding unit having a binding affinity for CD3, wherein the first binding unit comprises a heavy chain variable region comprising CDR1, CDR2 and CDR3 sequences in a human VH framework, wherein the CDR sequences comprise a sequence having two or less substitutions in a CDR sequence selected from the group consisting of SEQ ID NO: 1-23.

Aspects of the invention include multispecific binding compounds comprising a first binding unit having a binding affinity for CD22 and a second binding unit having a binding affinity for CD3, wherein the first binding unit comprises a heavy chain variable region comprising CDR1, CDR2 and CDR3 sequences in a human VH framework, wherein the CDR sequences are selected from the group consisting of SEQ ID NO: 1-23.

Aspects of the invention include multispecific binding compounds comprising a first binding unit having a binding affinity for CD22 and a second binding unit having a binding affinity for CD3, wherein the first binding unit comprises a heavy chain variable region comprising: (a) a CDR1 sequence of SEQ ID NO: 1, a CDR2 sequence of SEQ ID NO: 11 and a CDR3 sequence of SEQ ID NO: 18; or (b) a CDR1 sequence of SEQ ID NO: 1, a CDR2 sequence of SEQ ID NO: 12 and a CDR3 sequence of SEQ ID NO: 19; or (c) a CDR1 sequence of SEQ ID NO: 1, a CDR2 sequence of SEQ ID NO: 12 and a CDR3 sequence of SEQ ID NO: 20 in a human VH framework.

In some embodiments, the multispecific binding compound is bispecific. In some embodiments, the multispecific binding compound is in CAR-T format.

Aspects of the invention include multispecific binding compounds comprising: (i) a heavy chain variable region having a binding affinity for CD3, comprising a CDR1 sequence of SEQ ID NO: 85, a CDR2 sequence of SEQ ID NO: 86, and a CDR3 sequence of SEQ ID NO: 87 in a human VH framework; (ii) a light chain variable region comprising

- 3 048216 a CDR1 sequence of SEQ ID NO: 88, a CDR2 sequence of SEQ ID NO: 89 and a CDR3 sequence of SEQ ID NO: 90 in a human VL framework; and (iii) an antigen-binding domain of an anti-CD22 heavy chain-only antibody comprising a CDR1 sequence of SEQ ID NO: 1, a CDR2 sequence of SEQ ID NO: 11 and a CDR3 sequence of SEQ ID NO: 18 in a human VH framework.

Aspects of the invention include multispecific binding compounds comprising: (i) a heavy chain variable region having a binding affinity for CD3, comprising a CDR1 sequence of SEQ ID NO: 85, a CDR2 sequence of SEQ ID NO: 86 and a CDR3 sequence of SEQ ID NO: 87 in a human VH framework; (ii) a light chain variable region comprising a CDR1 sequence of SEQ ID NO: 88, a CDR2 sequence of SEQ ID NO: 89 and a CDR3 sequence of SEQ ID NO: 90 in a human VL framework; and (iii) an antigen-binding domain of an anti-CD22 heavy chain-only antibody comprising a CDR1 sequence of SEQ ID NO: 1, a CDR2 sequence of SEQ ID NO: 12 and a CDR3 sequence of SEQ ID NO: 19 in a human VH framework.

Aspects of the invention include a multispecific binding compound comprising: (i) a heavy chain variable region having a binding affinity for CD3, comprising a CDR1 sequence of SEQ ID NO: 85, a CDR2 sequence of SEQ ID NO: 86 and a CDR3 sequence of SEQ ID NO: 87 in a human VH framework; (ii) a light chain variable region comprising a CDR1 sequence of SEQ ID NO: 88, a CDR2 sequence of SEQ ID NO: 89 and a CDR3 sequence of SEQ ID NO: 90 in a human VL framework; and (iii) an antigen-binding domain of an anti-CD22 heavy chain-only antibody comprising a CDR1 sequence of SEQ ID NO: 1, a CDR2 sequence of SEQ ID NO: 12 and a CDR3 sequence of SEQ ID NO: 20 in a human VH framework.

In some embodiments, the multispecific binding compound comprises a human IgG1 Fc region. In some embodiments, the human IgG1 Fc region is a silenced human IgG1 Fc region. In some embodiments, the multispecific binding compound comprises a human IgG4 Fc region. In some embodiments, the human IgG4 Fc region is a silenced human IgG4 Fc region.

Aspects of the invention include pharmaceutical compositions comprising a multispecific binding compound as described herein.

Aspects of the invention include methods of treating a B-cell disorder characterized by CD22 expression, comprising administering to a subject with said disorder a multispecific binding compound or pharmaceutical composition as described herein.

Aspects of the invention include the use of a polyspecific binding compound in the preparation of a medicament for the treatment of a B-cell disorder characterized by CD22 expression.

In some embodiments, the disorder is diffuse large B-cell lymphoma (DLBCL). In some embodiments, the disorder is non-Hodgkin lymphoma (NHL). In some embodiments, the disorder is systemic lupus erythematosus (SLE). In some embodiments, the disease is rheumatoid arthritis (RA). In some embodiments, the disorder is multiple sclerosis (MS).

Aspects of the invention include polynucleotides encoding a multispecific binding compound as described herein. Aspects of the invention include vectors containing the polynucleotides as described herein. Aspects of the invention include cells containing the vectors described herein.

Aspects of the invention include methods for producing a multispecific binding compound as described herein, comprising growing a cell as described herein under conditions permitting expression of the binding compound, and isolating the binding compound from the cell.

Aspects of the invention include methods for producing a polyspecific binding compound as described herein, comprising immunizing an animal with UniRat CD22 and identifying CD22-binding heavy chain sequences.

Aspects of the invention include methods of treatment comprising administering to an individual an effective dose of a multispecific binding compound as described herein or a pharmaceutical composition as described herein.

These and other aspects will be further explained in the rest of the description, including examples.

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Brief description of graphic materials

Fig. 1A is a graph depicting T cell-mediated cytotoxicity of CD22-positive cells (Daudi) using resting human T cells.

Fig. 1B is a graph depicting the dose-response curves of cytokine release during resting human pan-T cells incubated with CD22-positive cells (Daudi) and treated with the polyspecific binding compound CD22xCD3_F2F and a positive control.

Fig. 2A is a graph depicting T cell-mediated cytotoxicity of CD22-positive cells (SUDHL10) using resting human pan-T cells.

Fig. 2B is a graph depicting the dose-response curves of cytokine release from resting human pan-T cells incubated with CD22-positive cells (SUDHL10) and treated with the anti-CD22xCD3_F2F polyspecific binding compound and a positive control.

Fig. 3A shows a series of graphs depicting T cell-mediated cytotoxicity of CD22-positive cells (RI-1) using resting human T cells.

Figure 3B shows a series of graphs depicting dose-response curves of cytokine release from resting human pan-G cells incubated with CD22-positive cells (RI-1) and treated with the polyspecific binding compound anti-CD22xCD3_F2F and a positive control.

Figure 4 shows a series of graphs depicting T cell-mediated cytotoxicity of CD22-positive cells using activated human pan-T cells.

Figure 5 shows a series of graphs depicting cell binding of bispecific antibodies against CD22 and CD3.

Fig. 6 shows the treatment plan to determine the in vivo efficacy of the anti-CD22xCD3_F2F polyspecific binding compound in Daudi xenografts.

Fig. 7 is a graph depicting the mean tumor volume as a function of days after tumor implantation in the Daudi xenograft mouse model.

Fig. 8 shows a graph depicting body weight as a function of days after tumor implantation in the Daudi xenograft mouse model.

Figure 9 is a graph depicting the percentage change in body weight as a function of days after tumor implantation in the Daudi xenograft mouse model.

Fig. 10 is a graph depicting the mean tumor volume as a function of days after tumor implantation in the Daudi xenograft mouse model.

Figure 11 shows a series of graphs depicting individual tumor measurements as a function of days after tumor implantation in the Daudi xenograft mouse model.

Fig. 12 is a graph depicting body weight as a function of days after tumor implantation in the Daudi xenograft mouse model.

Fig. 13 is a graph depicting the percentage change in body weight as a function of days after tumor implantation in the Daudi xenograft mouse model.

Fig. 14A is a schematic of a bispecific binding compound having one binding unit that specifically binds to CD3 and one binding unit that specifically binds to CD22.

Fig. 14B provides illustrations of various CAR-T constructs that may include one or more binding domains in accordance with embodiments of the present invention.

Fig. 15A is a schematic of a bispecific binding molecule having one binding unit that specifically binds to CD3 and one binding unit that specifically binds to CD22 (monovalent, monospecific for CD22).

Fig. 15B shows a schematic of a bispecific binding molecule having one binding unit that specifically binds to CD3 and two binding units that specifically bind to CD22 (bivalent, monospecific for CD22).

Fig. 15C is a schematic of a bispecific binding molecule having one binding unit that specifically binds to CD3 and two binding units that specifically bind to CD22 (bivalent, biparatopic with respect to CD22).

Fig. 16 is a table showing data for various biological activities of CD22 antibodies according to embodiments of the invention.

Fig. 17 shows a series of graphs showing the serum titer as a function of dilution.

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Detailed Description of Preferred Embodiments

The practice of the present invention will employ, unless otherwise indicated, conventional techniques of molecular biology (including recombinant techniques), microbiology, cell biology, biochemistry, and immunology, which are within the skill of the art. Such techniques are fully described in the literature, for example, in Molecular Cloning: A Laboratory Manual, second edition (Sambrook et al., 1989); Oligonucleotide Synthesis (M. J. Gait, ed., 1984); Animal Cell Culture (R. I. Freshney, ed., 1987); Methods in Enzymology (Academic Press, Inc.); Current Protocols in Molecular Biology (F. M. Ausubel et al., eds., 1987, and periodic updates); PCR: The Polymerase Chain Reaction, (Mullis et al., ed., 1994); A Practical Guide to Molecular Cloning (Perbal Bernard V., 1988); Phage Display: A Laboratory Manual (Barbas et al., 2001); Harlow, Lane and Harlow, Using Antibodies: A Laboratory Manual: Portable Protocol No. I, Cold Spring Harbor Laboratory (1998); and Harlow and Lane, Antibodies: A Laboratory Manual, Cold Spring Harbor Laboratory; (1988).

When a range of values is given, it is to be understood that the invention covers each intermediate value, to the accuracy of the tenth decimal place of the lower limit, unless the context clearly indicates otherwise, between the upper and lower limits of this range, as well as any other stated or intermediate value in such stated range. The upper and lower limits of these smaller ranges may be independently included in the smaller ranges and are also included in the invention, taking into account any specifically excluded limit in the stated range. If a stated range includes one or both limits, ranges excluding one or both of these included limits are also included in the invention.

Unless otherwise noted, antibody residues in this document are numbered according to the Kabat numbering system (e.g., Kabat et al., Sequences of Immunological Interest. 5th Ed. Public Health Service, National Institutes of Health, Bethesda, Md. (1991)).

In order to provide a thorough understanding of the present invention, numerous specific details are set forth in the following detailed description. However, it will be apparent to one skilled in the art that the present invention may be practiced without one or more of these specific details. In other instances, well-known features and procedures well known to those skilled in the art have not been described in order to avoid obscuring the invention.

All references cited in this document, including patent applications and publications, are incorporated herein by reference in their entirety.

I. Definitions.

The term “comprising” means that the listed elements are required for the composition/method/kit, but other elements may be included within the scope of the claims to form the composition/method/kit and the like.

The term "consisting essentially of" means limiting the scope of the described composition or method to the specified materials or stepwise actions that do not materially affect the basic and novel characteristic(s) of the subject matter of the invention.

The term "consisting of" means the exclusion of any element, step, or ingredient not specified in the claims from the composition, method, or kit.

Antibody residues herein are numbered according to the Kabat numbering system and the EU numbering system. The Kabat numbering system is generally used to refer to a residue in the variable domain (approximately residues 1-113 of the heavy chain) (e.g., Kabat et al., Sequences of Immunological Interest. 5th Ed. Public Health Service, National Institutes of Health, Bethesda, Md. (1991)). The EU numbering system or EU index is generally used to refer to an amino acid residue in the constant region of an immunoglobulin heavy chain (e.g., the EU index presented in Kabat et al., supra). The EU index of Kabat refers to the amino acid residue numbering of the human IgG1 antibody EU. Unless otherwise noted, references to residue numbers in the variable domain of antibodies are to residue numbering according to the Kabat numbering system. Unless otherwise noted, references to residue numbers in the constant domain of antibodies are to residue numbering according to the EU numbering system.

Antibodies, also called immunoglobulins, typically comprise at least one heavy chain and one light chain, wherein the amino-terminal domain of the heavy and light chains is variable in sequence, so it is typically called the variable region domain or the variable domain of the heavy chain (VH) or the variable domain of the light chain (VH). These two domains typically associate to form a specific binding region, although, as will be discussed herein, specific binding can also be obtained with variable sequences of only the heavy chains, and various non-natural antibody configurations are known and used in the art.

A functional or biologically active antibody or antigen-binding molecule (including heavy chain-only antibodies and polyspecific (e.g., bispecific) three-chain antibody-like (TCA) molecules as described herein) is a molecule capable of exerting one or more of its natural activities in structural,

- 6 048216 regulatory, biochemical, or biophysical events. For example, a functional antibody or other binding molecule, such as TCA, may have the ability to specifically bind an antigen, and the binding may in turn induce or alter a cellular or molecular event, such as signal transduction or enzymatic activity. A functional antibody or other binding molecule, such as TCA, may also block ligand activation of a receptor or act as an agonist or antagonist. The ability of an antibody or other binding molecule, such as TCA, to exhibit one or more of its natural activities depends on several factors, including proper folding and assembly of the polypeptide chains.

The term antibody is used in the broadest sense herein and specifically encompasses monoclonal antibodies, polyclonal antibodies, monomers, dimers, multimers, polyspecific antibodies (e.g., bispecific antibodies), heavy chain-only antibodies, three-chain antibodies, single-chain Fv (scFv), nanobodies, etc., and also includes antibody fragments if they exhibit the desired biological activity (Miller et al (2003) Jour, of Immunology 170:4854-4861). Antibodies may be murine, human, humanized, chimeric, or derived from other species.

The term "antibody" may refer to a full-length heavy chain, a full-length light chain, an intact immunoglobulin molecule; or an immunologically active portion of any of these polypeptides, i.e., a polypeptide that contains an antigen-binding site that immunospecifically binds an antigen of a target of interest or a portion thereof, such targets including, but not limited to, cancer cells or cells that produce autoimmune antibodies associated with an autoimmune disease. The immunoglobulin described herein may be any type (e.g., IgG, IgE, IgM, IgD, and IgA), class (e.g., IgG1, IgG2, IgG3, IgG4, IgAl, and IgA2), or subclass of an immunoglobulin molecule, including engineered subclasses with altered Fc portions that provide for a decrease or increase in effector cell activity. Immunoglobulins may be derived from any species. In one aspect, the immunoglobulin is predominantly of human origin.

The term monoclonal antibody, as used herein, refers to an antibody obtained from a population of substantially homogeneous antibodies, i.e., the individual antibodies in the population are identical except for naturally occurring mutations that may be present in small numbers. Monoclonal antibodies are highly specific and are directed against a single antigenic site. Furthermore, unlike conventional (polyclonal) antibody preparations, which typically include different antibodies directed against different determinants (epitopes), each monoclonal antibody is directed against a single determinant on the antigen. Monoclonal antibodies according to the present invention can be prepared by the hybridoma method first described by Kohler et al. (1975) Nature 256:495, and can also be prepared, for example, by recombinant protein production methods (see, e.g., U.S. Patent No. 4,816,567).

The term variable, as used herein in connection with antibodies, refers to the fact that the sequences of some portions of the variable domains of an antibody vary widely among antibodies and contribute to the binding and specificity of each particular antibody for its particular antigen. However, variability is not uniformly distributed across the variable domains of antibodies. It is concentrated in three segments of the variable domains of both the light and heavy chains, called hypervariable regions. The more conserved portions of the variable domains are called framework regions (FRs). The variable domains of native light and heavy chains each contain four FRs, predominantly adopting a β-sheet configuration, and connected by three hypervariable regions that form loops connecting, and in some cases are part of, the β-type structures. The hypervariable regions of each chain are joined together in close proximity by FRs and, together with the hypervariable regions of the other chain, participate in the formation of the antigen-binding site of antibodies (see Kabat et al., Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, MD (1991)). The constant domains are not directly involved in binding of the antibody to antigen, but they exhibit various effector functions, such as participation of the antibody in antibody-dependent cell-mediated cytotoxicity (ADCC).

The term hypervariable region, as used herein, refers to the amino acid residues of an antibody that are responsible for antigen binding. The hypervariable region typically comprises amino acid residues from the complementarity determining region or CDR (e.g., residues 31-35 (H1), 50-65 (H2), and 95-102 (H3) in the variable domain of the heavy chain; Kabat et al., Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, MD. (1991)) and/or residues from the hypervariable loop residues 26-32 (H1), 53-55 (H2), and 96-101 (H3) in the variable domain of the heavy chain; Chothia and Lesk J. Mol. Biol. 196:901-917 (1987)). Framework region or FR residues, as defined herein, are hypervariable region residues distinct from variable domain residues.

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Exemplary CDR designations are shown herein, however, one skilled in the art will appreciate that a number of CDR definitions are commonly used, including the Kabat definition (see Zhao et al. A germline knowledge based computational approach for determining antibody complementarity determining regions. Mol Immunol. 2010; 47:694-700), which is based on sequence variability and is the most commonly used. The Chothia definition is based on the arrangement of structural loop regions (Chothia et al. Conformations of immunoglobulin hypervariable regions. Nature. 1989; 342:877-883). Alternative CDR definitions of interest include, but are not limited to, those disclosed by Honegger, Yet another numbering scheme for immunoglobulin variable domains: an automatic modeling and analysis tool. J Mol Biol. 2001; 309:657-670; Ofran et al. Automated identification of complementarity determining regions (CDRs) reveals peculiar characteristics of CDRs and B cell epitopes. J Immunol. 2008; 181:6230–6235; Almagro Identification of differences in the specificity-determining residues of antibodies that recognize antigens of different sizes: implications for the rational design of antibody repertoires. J Mol Recognit. 2004; 17:132–143; and Padlanet al. Identification of specificity-determining residues in antibodies. Faseb J. 1995; 9:133–139, the contents of each of which are incorporated herein by reference in their entirety.

The terms heavy chain-only antibody, heavy chain antibody are used interchangeably herein and refer in the broadest sense to antibodies that lack the light chain of a conventional antibody. The terms specifically include, but are not limited to, homodimeric antibodies comprising a VH antigen-binding domain and CH2 and CH3 constant domains, in the absence of a CH1 domain; functional (antigen-binding) variants of such antibodies, soluble VH variants, Ig-NARs comprising a homodimer of one variable domain (V-NAR) and five C-like constant domains (C-NAR), and functional fragments thereof; and soluble single-domain antibodies (sUniDab™). In one embodiment, the heavy chain-only antibody consists of an antigen-binding domain of a variable region consisting of framework 1, CDR1, framework 2, CDR2, framework 3, CDR3, and framework 4. In another embodiment, the heavy chain-only antibody consists of an antigen-binding domain of at least a portion of a hinge region and the CH2 and CH3 domains. In another embodiment, the heavy chain-only antibody consists of an antigen-binding domain of at least a portion of a hinge region and a CH2 domain. In a further embodiment, the heavy chain-only antibody consists of an antigen-binding domain of at least a portion of a hinge region and a CH3 domain. Heavy chain-only antibodies in which the CH2 and/or CH3 domain is truncated are also included herein. In a further embodiment, the heavy chain consists of an antigen-binding domain and at least one CH domain (CH1, CH2, CH3 or CH4), but without a hinge region. In a further embodiment, the heavy chain consists of an antigen-binding domain, at least one CH domain (CH1, CH2, CH3 or CH4) and at least part of a hinge region. The heavy chain-only antibody may be in the form of a dimer in which two heavy chains are disulfide-linked to each other, covalently or non-covalently linked to each other. The heavy chain-only antibody may belong to the IgG subclass, but antibodies belonging to other subclasses, such as the IgM, IgA, IgD and IgE subclass are also included herein. In a particular embodiment, the heavy chain antibody has an IgG1, IgG2, IgG3 or IgG4 subtype, in particular an IgG1 subtype. In one embodiment, the heavy chain antibody is of the IgG4 subtype, wherein one or more CH domains are modified to alter the effector function of the antibody. In one embodiment, the heavy chain antibody is of the IgG1 subtype, wherein one or more CH domains are modified to alter the effector function of the antibody. Modifications of the CH domains that alter effector function are further described herein. Non-limiting examples of heavy chain antibodies are described, for example, in WO2018/039180, the disclosure of which is incorporated herein by reference in its entirety.

In one embodiment, heavy chain-only antibodies are used herein as the binding (targeting) domain of a chimeric antigen receptor (CAR). The definition specifically includes human heavy chain-only antibodies, referred to as UniAbs™, produced by human immunoglobulin transgenic rats (UniRat™). The variable regions (VH) of UniAbs™ are referred to as UniDab™, and are versatile building blocks that can be linked to Fc regions or serum albumin to develop new therapeutics with polyspecificity, increased potency and extended half-life. Since homodimeric UniAbs™ lack a light chain and therefore a VL domain, the antigen is recognized by a single domain, i.e., the variable domain (antigen binding domain) of the heavy chain of the heavy chain antibody (VH).

As used herein, the term intact antibody chain is a chain that contains a full-length variable region and a full-length constant region (Fc). An intact reference antibody contains an intact light chain and an intact heavy chain, as well as a light chain constant domain (CL) and heavy chain constant domains, CH1, hinge, CH2 and CH3

- 8 048216 for secreted IgG. Other isotypes, such as IgM or IgA, may have different CH domains. The constant domains may be native sequence constant domains (e.g., human native sequence constant domains) or amino acid sequence variants thereof. An intact antibody may have one or more effector functions, which refer to the biological activity that is inherent in the Fc constant region (either a native sequence Fc region or an amino acid sequence variant Fc region) of the antibody. Examples of antibody effector functions include C1q binding; complement-dependent cytotoxicity; Fc receptor binding; antibody-dependent cell-mediated cytotoxicity (ADCC); phagocytosis; and downregulation of cell surface receptors. Constant region variants include those that alter the effector profile, Fc receptor binding, etc.

Depending on the amino acid sequence of the Fc (constant domain) of their heavy chains, antibodies and various antigen-binding proteins can be classified into different classes. There are five major classes of heavy chain Fc regions: IgA, IgD, IgE, IgG, and IgM, and some of these can be further divided into subclasses (isotypes), such as IgG1, IgG2, IgG3, IgG4, IgA, and IgA2. The Fc constant domains that correspond to the different classes of antibodies can be designated as α, δ, ε, γ, and μ, respectively. The subunit structures and three-dimensional configurations of the different immunoglobulin classes are well known. Ig forms include hinged modifications or hingeless forms (Roux et al (1998) J. Immunol. 161:4083–4090; Lund et al (2000) Eur. J. Biochem. 267:7246–7256; US 2005/0048572; US 2004/0229310). The light chains of antibodies from any vertebrate species can be classified into one of two types, called μ and λ, based on the amino acid sequences of their constant domains.

The functional Fc region has an effector function of the native sequence Fc region. Non-limiting examples of effector functions include C1q binding; CDC; Fc receptor binding; ADCC; ADCF; down-regulation of cell surface receptors (e.g., B cell receptor), etc. Such effector functions typically require the Fc region to interact with a receptor, such as the FcγRI; FcγRIIA; FcγRIIBI; FcγRIIB2; FcγRIIIA; FcγRIIIB receptors, and the low affinity FcRn receptor; and can be assessed using a variety of assays known in the art. A dead or silenced Fc is an Fc that has been mutated to retain activity, such as prolonging serum half-life, but which does not activate the high-affinity Fc receptor or which has reduced affinity for the Fc receptor.

A native sequence Fc region comprises an amino acid sequence identical to the amino acid sequence of an Fc region found in nature. Native sequence human Fc regions include, for example, a native sequence human IgG1 Fc region (non-A and A allotypes); a native sequence human IgG2 Fc region; a native sequence human IgG3 Fc region; and a native sequence human IgG4 Fc region, as well as naturally occurring variants thereof.

A variant Fc region comprises an amino acid sequence that differs from the sequence of a native sequence Fc region by at least one amino acid modification, preferably one or more amino acid substitutions. Preferably, the variant Fc region has at least one amino acid substitution compared to the native sequence Fc region or the Fc region of the parent polypeptide, such as from about one to about ten amino acid substitutions, and preferably from about one to about five amino acid substitutions in the native sequence Fc region or the Fc region of the parent polypeptide. A variant Fc region herein will preferably have at least about 80% homology with the native sequence Fc region and/or the Fc region of the parent polypeptide, and most preferably at least about 90% homology therewith, more preferably at least about 95% homology therewith.

Fc sequence variants may contain three amino acid substitutions in the CH2 region to reduce FcγRI binding at EU positions 234, 235, and 237 (see Duncan et al., (1988) Nature 332:563). Two amino acid substitutions in the complement-fixing site C1q at EU positions 330 and 331 reduce complement fixation (see Tao et al., J. Exp. Med. 178:661 (1993) and Canfield and Morrison, J. Exp. Med. 173:1483 (1991)). Substitution with human IgG1 or IgG2 residues at positions 233–236 and IgG4 residues at positions 327, 330, and 331 significantly reduces ADCC and CDC (see, e.g., Armour KL et al, 1999 Eur J Immunol. 29(8):2613–24; and Shields RL et al, 2001. J Biol Chem. 276(9):6591–604). The amino acid sequence of human IgG1 (UniProtKB No. P01857) is provided herein as SEQ ID NO: 93. The amino acid sequence of human IgG4 (UniProtKB No. P01861) is provided herein as SEQ ID NO: 94. Silenced IgG1 is described, for example, in Boesch, A.W., et al., Highly parallel characterization of IgG Fc binding interactions. MAbs, 2014. 6(4): p. 915-27, the disclosure of which is incorporated herein in its entirety.

- 9 048216 via link.

Other Fc variants are possible, including, but not limited to, a variant in which a region capable of forming a disulfide bond is removed, or in which certain amino acid residues at the N-terminus of a native Fc are removed, or a methionine residue is added thereto. Thus, in some embodiments, one or more Fc portions of a binding compound may comprise one or more mutations in the hinge region to eliminate a disulfide bond. In another embodiment, the hinge region of an Fc may be completely removed. In another embodiment, a binding compound may comprise an Fc variant.

In addition, an Fc variant can be engineered to remove or substantially reduce effector functions by substitution (mutation), deletion, or addition of amino acid residues for complement binding or Fc receptor binding. For example, without limitation, the deletion can occur in a complement binding site, such as a C1q binding site. Methods for producing such immunoglobulin Fc fragment sequence derivatives are described in International Patent Publication Nos. WO 97/34631 and WO 96/32478. In addition, the Fc domain can be modified by phosphorylation, sulfation, acylation, glycosylation, methylation, farnesylation, acetylation, amidation, and the like.

The term "antibody comprising an Fc region" refers to an antibody that comprises an Fc region. The C-terminal lysine (residue 447 according to the EU numbering system) of the Fc region may be removed, for example, during purification of the antibody or by constructing a recombinant nucleic acid encoding the antibody. Accordingly, an antibody having an Fc region according to the present invention may include an antibody with or without K447.

Aspects of the invention include binding compounds having polyspecific configurations, which include, without limitation, bispecific, trispecific, etc. A wide variety of methods and protein configurations are known and used to produce bispecific monoclonal antibodies (bSMAbs), trispecific antibodies, etc.

Various methods have been developed for producing polyvalent artificial antibodies by recombinantly fusing the variable domains of two or more antibodies. In some embodiments, the first and second antigen-binding domains on the polypeptide are connected by a polypeptide linker. One non-limiting example of such a polypeptide linker is a GS linker having an amino acid sequence of four glycine residues followed by one serine residue, and in which the sequence is repeated n times, where n is an integer from 1 to about 10 (SEQ ID NO: 107), such as 2, 3, 4, 5, 6, 7, 8, or 9. Non-limiting examples of such linkers include GGGGS (SEQ ID NO: 102) (n=1) and GGGGSGGGGS (SEQ ID NO: 103) (n=2). Other suitable linkers can also be used, as described in, for example, Chen et al., Adv Drug Deliv Rev. 2013 October 15; 65(10):1357-69, the disclosure of which is incorporated herein by reference in its entirety.

The term three-chain antibody-like molecule or TCA is used herein to refer to antibody-like molecules comprising, consisting essentially of, or consisting of three polypeptide subunits, two of which comprise, consist essentially of, or consist of one heavy and one light chain of a monoclonal antibody, or functional antigen-binding fragments of such antibody chains, comprising an antigen-binding region and at least one CH domain. The heavy chain/light chain pair has binding specificity for a first antigen. The third polypeptide subunit comprises, consists essentially of, or consists of a heavy chain-only antibody comprising an Fc portion comprising CH2 and/or CH3 and/or CH4 domains, in the absence of a CH1 domain, and one or more antigen-binding domains (e.g., two antigen-binding domains) that bind an epitope of a second antigen or a different epitope of a first antigen, wherein such binding domain is derived from or has sequence identity with a variable region of a heavy or light chain of the antibody. Portions of such a variable region may be encoded by VH and/or VL gene segments, D and JH gene segments, or JL gene segments. The variable region may be encoded by rearranged VHDJH, VLDJH, VHJL, or V _L J _L gene segments. The TCA protein utilizes a heavy chain-only antibody as defined above.

The TCA binding compound utilizes a heavy chain-only antibody or an antibody consisting of heavy chains or a heavy chain-only polypeptide, which as used herein means a single chain antibody comprising heavy chain constant regions of CH2 and/or CH3 and/or CH4 but without the CH1 domain. In one embodiment, the heavy chain-only antibody consists of an antigen-binding domain, at least a portion of a hinge region, and the CH2 and CH3 domains. In another embodiment, the heavy chain-only antibody consists of an antigen-binding domain, at least a portion of a hinge region, and the CH2 domain. In a further embodiment, the heavy chain-only antibody consists of an antigen-binding domain, at least a portion of a hinge region, and the CH3 domain. Heavy chain-only antibodies in which the CH2 and/or CH3 domain is truncated are also included herein. In another embodiment, the heavy chain consists of an antigen-binding

- 10 048 216 a hinge domain and at least one CH domain (CH1, CH2, CH3 or CH4), but without a hinge region. The heavy chain-only antibody may be in the form of a dimer in which two heavy chains are disulfide bonded, the others are otherwise covalently or non-covalently linked to each other, and may optionally include an asymmetric interaction interface between two or more CH domains to facilitate proper pairing between the polypeptide chains. The heavy chain antibody may belong to the IgG subclass, but antibodies belonging to other subclasses such as the IgM, IgA, IgD and IgE subclass are also included. In a particular embodiment, the heavy chain-only antibody is of the IgG1, IgG2, IgG3 or IgG4 subtype, in particular the IgG1 subtype or the IgG4 subtype. Non-limiting examples of TCA binding compounds are described, for example, in WO2017/223111 and WO2018/052503, the disclosures of which are incorporated herein by reference in their entirety.

Heavy-chain-only antibodies account for about a quarter of the IgG antibodies produced by camelids such as camels and llamas (Hamers-Casterman C., et al. Nature. 363, 446-448 (1993)). These antibodies are formed by two heavy chains but lack light chains. As a consequence, the variable antigen-binding region is called the VHH domain, and it is the smallest naturally occurring, intact antigen-binding site, being only about 120 amino acids long (Desmyter, A., et al. J. Biol. Chem. 276, 26285-26290 (2001)). Heavy chain-only antibodies with high specificity and affinity can be raised against a variety of antigens by immunization (van der Linden, R. H., et al. Biochim. Biophys. Acta. 1431, 37-46 (1999)), and the VHH portion can be readily cloned and expressed in yeast (Frenken, L. G. J., et al. J. Biotechnol. 78, 11-21 (2000)). Their expression levels, solubility, and stability are significantly higher than those of classical F(ab) or Fv fragments (Ghahroudi, M. A. et al. FEBS Lett. 414, 521-526 (1997)). Sharks have also been shown to have a single VH-like domain in their antibodies, called VNAR. (Nuttall et al. Eur. J. Biochem. 270, 3543-3554 (2003); Nuttall et al. Function and Bioinformatics 55, 187-197 (2004); Dooley et al., Molecular Immunology 40, 25-33 (2003)).

As used herein, the terms CD22 and cluster of differentiation 22 refer to a molecule belonging to the SIGLEC family of lectins found on the surface of mature B cells and, to a lesser extent, on some immature B cells. The term CD22 includes the CD22 protein of humans and any non-human animal species, and in particular includes human CD22 as well as CD22 of non-human mammals.

As used herein, the term human CD22 includes any variants, isoforms, and species homologues of human CD22 (UniProt P20273), regardless of its source or method of production. Thus, human CD22 includes human CD22 naturally expressed by cells and CD22 expressed on cells transfected with the human CD22 gene.

The terms "anti-CD22 heavy chain only antibody," "anti-CD22 heavy chain only antibody," "anti-CD22 heavy chain only antibody," and "anti-CD22 heavy chain only antibody" are used interchangeably herein to refer to a heavy chain only antibody as defined above for immunospecifically binding to CD22, including human CD22 as defined above. The definition includes, but is not limited to, human heavy chain only antibodies produced by transgenic animals, such as transgenic rats or transgenic mice expressing human immunoglobulin, including UniRat™ producing human UniAb™ antibodies to CD22 as defined above.

The percentage (%) identity of amino acid sequences relative to a reference polypeptide sequence is defined as the percentage of amino acid residues in a candidate sequence that are identical to amino acid residues in the reference polypeptide sequence, after aligning the sequences and introducing gaps, if necessary, to achieve maximum sequence identity, but without considering any conservative substitutions as part of the sequence identity. Alignment for the purpose of determining percent amino acid sequence identity can be accomplished in a variety of ways known to those skilled in the art, for example, using publicly available computer programs such as BLAST, BLAST2, ALIGN, or Megalign (DNASTAR) software. Those skilled in the art can determine appropriate parameters for aligning sequences, including any algorithms necessary to achieve maximum alignment over the entire length of the sequences being compared. For the purposes of this document, however, the % amino acid sequence identity values are obtained using the ALIGN-2 sequence comparison program.

An isolated antibody is one that has been identified and separated and/or recovered from a component of its natural environment. A contaminant component of the natural environment is a substance that interferes with the diagnostic or therapeutic use of the antibody, and may include enzymes, hormones, and other proteinaceous or non-proteinaceous solutes. In preferred embodiments, the antibody will be purified (1) to greater than 95% by weight of the antibody as determined by the Lowry method, and most preferably greater than 99% by weight, (2) to a degree

- 11 048216 sufficient to obtain at least 15 residues of the N-terminal or internal amino acid sequence using a spinning cup sequencer, or (3) to homogeneity by SDS-PAGE under reducing or non-reducing conditions using Coomassie blue or, preferably, a silver-based stain. Isolated antibody includes antibody in situ in recombinant cells since at least one component of the antibody's natural environment will be missing. Typically, however, isolated antibody will be obtained by at least one purification step.

The antibodies of the invention include multispecific antibodies. Multispecific antibodies have more than one binding specificity. The term multispecific particularly includes bispecific and trispecific, as well as higher order independent specific binding affinity, such as higher order polyepitope specificity, as well as tetravalent antibodies and antibody fragments. The terms multispecific antibody, multispecific heavy chain-only antibody, multispecific heavy chain antibody, multispecific UniAb™ and multispecific binding compound are used herein in the broadest sense and cover all antibodies with more than one binding specificity. The heavy chain-only multispecific CD22 antibodies of the present invention particularly include antibodies that immunospecifically bind to one single epitope on a CD22 protein, such as human CD22, and to an epitope on another protein, such as, for example, the CD3 protein (i.e., bivalent and monoparatopic). The heavy chain-only multispecific CD22 antibodies of the present invention particularly include antibodies that immunospecifically bind to two or more non-overlapping epitopes on a CD22 protein, such as human CD22 (i.e., bivalent and biparatopic). The heavy chain-only multispecific anti-CD22 antibodies of the present invention also specifically include antibodies that immunospecifically bind to an epitope of a CD22 protein, such as human CD22, and an epitope of another protein, such as, for example, a CD3 protein, such as human CD3 (i.e., bivalent and biparatopic). The heavy chain-only multispecific anti-CD22 antibodies of the present invention also specifically include antibodies that immunospecifically bind to two or more non-overlapping or partially overlapping epitopes on a CD22 protein, such as human CD22 protein, and an epitope on another protein, such as, for example, a CD3 protein, such as human CD3 protein (i.e., trivalent and biparatopic).

The antibodies of the invention include monospecific antibodies having a single binding specificity. Monospecific antibodies particularly include antibodies having a single binding specificity as well as antibodies comprising more than one binding unit having the same binding specificity. The terms monospecific antibody, monospecific heavy chain-only antibody, monospecific heavy chain antibody and monospecific UniAb™ are used herein in the broadest sense and encompass all antibodies having a single binding specificity. The monospecific heavy chain-only anti-CD22 antibodies of the invention particularly include antibodies that immunospecifically bind to a single epitope of the CD22 protein, such as human CD22 (monovalent and monospecific). Monospecific heavy chain-only anti-CD22 antibodies of the present invention also specifically include antibodies having more than one binding unit (e.g., multivalent antibodies) that immunospecifically bind to an epitope on a CD22 protein, such as human CD22. For example, a monospecific antibody according to embodiments of the invention may comprise a heavy chain variable region comprising two antigen-binding domains, wherein each antigen-binding domain binds to the same epitope on a CD22 protein (i.e., bivalent and monospecific).

An epitope is a site on the surface of an antigen molecule to which one antibody molecule binds. Typically, an antigen has several or many different epitopes and reacts with many different antibodies. The term specifically includes linear epitopes and conformational epitopes.

Epitope mapping is the process of identifying the binding sites, or epitopes, of antibodies on their target antigens. Antibody epitopes can be linear or conformational epitopes. Linear epitopes are formed by a continuous sequence of amino acids in a protein. Conformational epitopes are formed by amino acids that are discontinuous in the protein sequence but that come together as the protein folds into its three-dimensional structure.

The term polyepitopic specificity refers to the ability to specifically bind to two or more different epitopes on the same or different targets. As noted above, the present invention particularly includes heavy chain-only anti-CD22 antibodies with polyepitopic specificity, i.e., heavy chain-only anti-CD22 antibodies that bind to one or more non-overlapping epitopes on a CD22 protein, such as human CD22; and anti-CD22 antibodies that bind to one or more epitopes on a CD22 protein and to an epitope on another protein, such as

- 12 048216 such as the CD3 protein. The term non-overlapping epitope(s) or non-competitive epitope(s) of an antigen is defined herein to mean epitope(s) that are recognized by one member of a pair of antigen-specific antibodies but not by the other member. Antibody pairs or antigen-binding regions targeting the same antigen on a polyspecific antibody that recognize non-overlapping epitopes do not compete for binding to that antigen and are capable of simultaneously binding that antigen.

An antibody binds substantially the same epitope as a reference antibody when the two antibodies recognize identical or sterically overlapping epitopes. The most widely used and rapid methods for determining whether two epitopes bind to identical or sterically overlapping epitopes are competitive binding assays, which can be configured in a variety of formats using either labeled antigen or labeled antibody. Typically, the antigen is immobilized in a 96-well plate, and the ability of unlabeled antibodies to block binding of labeled antibodies is measured using radioactive or enzymatic labels.

The term valence, as used herein, refers to the specified number of binding sites in an antibody molecule.

A monovalent antibody has one binding site. Thus, a monovalent antibody is also monospecific.

A multivalent antibody has two or more binding sites. Thus, the terms bivalent, trivalent and tetravalent refer to the presence of two binding sites, three binding sites and four binding sites, respectively. Thus, a bispecific antibody of the invention is at least bivalent and may be trivalent, tetravalent or otherwise multivalent. A bivalent antibody according to embodiments of the invention may have two binding sites with the same epitope (i.e., bivalent, monoparatopic) or with two different epitopes (i.e., bivalent, biparatopic).

A wide variety of protein methods and configurations are known and are used to produce bispecific monoclonal antibodies (bSMAbs), trispecific antibodies, and the like.

The term chimeric antigen receptor or CAR is used herein in its broadest sense to refer to an engineered receptor that grafts a desired binding specificity (e.g., the antigen-binding region of a monoclonal antibody or other ligand) onto membrane-spanning and intracellular signaling domains. Typically, the receptor is used to graft the specificity of a monoclonal antibody onto a T cell to create a chimeric antigen receptor (CAR). (J Natl Cancer Inst 2015; 108(7):dvj439; and Jackson et al. 2016; 13:370–383).

The term human antibody is used herein to include antibodies having variable and constant regions derived from human germline immunoglobulin sequences. Human antibodies herein may include amino acid residues that are not encoded by human germline immunoglobulin sequences, such as mutations introduced by random or site-directed mutagenesis in vitro or by somatic mutation in vivo. The term human antibody particularly includes heavy chain-only antibodies having human heavy chain variable region sequences produced by transgenic animals, such as transgenic rats or mice, in particular UniAb™ produced by UniRat™ as defined above.

By chimeric antibody or chimeric immunoglobulin is meant an immunoglobulin molecule comprising amino acid sequences from at least two different Ig loci, for example, a transgenic antibody comprising a portion encoded by a human Ig locus and a portion encoded by a rat Ig locus. Chimeric antibodies include transgenic antibodies with non-human Fc regions or artificial Fc regions, and human idiotypes. Such immunoglobulins can be isolated from animals according to the invention that have been engineered to produce such chimeric antibodies.

As used herein, the term effector cell refers to an immune cell that participates in the effector phase of an immune response, as opposed to the cognitive and activation phases of an immune response. Some effector cells express specific Fc receptors and perform specific immune functions. In some embodiments, an effector cell, such as a natural killer cell, is capable of inducing antibody-dependent cellular cytotoxicity (ADCC). For example, monocytes and macrophages that express FcRs participate in the specific killing of target cells and the presentation of antigens to other components of the immune system or binding to cells that present antigens. In some embodiments, an effector cell can phagocytize a target antigen or a target cell.

Human effector cells are leukocytes that express receptors such as T cell receptors or FcRs and perform effector functions. Preferably,

- 13 048216 such cells express at least FcyRIII and perform ADCC effector function. Examples of human leukocytes that mediate ADCC include natural killer (NK) cells, monocytes, cytotoxic T cells, and neutrophils; NK cells are preferred. Effector cells can be isolated from a native source, such as blood or PBMC, as described herein.

The term immune cell is used herein in its broadest sense to include, without limitation, cells of myeloid or lymphoid origin, such as lymphocytes (such as B cells and T cells, including cytolytic T cells (CTLs)), natural killer (NK) cells, macrophages, monocytes, eosinophils, polymorphonuclear cells such as neutrophils, granulocytes, mast cells, and basophils.

Antibody effector functions refer to those biological activities that are attributable to the Fc region (either the native sequence Fc region or the variant amino acid sequence Fc region) of the antibody. Examples of antibody effector functions include C1q binding; complement-dependent cytotoxicity (CDC); Fc receptor binding; antibody-dependent cell-mediated cytotoxicity (ADCC); phagocytosis; downregulation of cell surface receptors (e.g., B cell receptor, BCR), etc.

Antibody-dependent cell-mediated cytotoxicity and ADCC refer to a cell-mediated response in which nonspecific cytotoxic cells that express Fc receptors (FcRs) (e.g., natural killer (NK) cells, neutrophils, and macrophages) recognize bound antibody on a target cell and thereby cause lysis of the target cell. The primary cells that mediate ADCC, natural killer cells, express only FcγRIII, whereas monocytes express FcγRI, FcγRII, and FcγRIII. FcγR expression on hematopoietic cells is summarized in Table 3 on page 464 of Ravetch and Kinet, Annu. Rev. Immunol 9:457-92 (1991). To assess the ADCC activity of a molecule of interest, an in vitro ADCC assay as described in U.S. Patent No. 5,500,362 or 5,821,337 can be performed. Effector cells suitable for such an assay include peripheral blood mononuclear cells (PBMCs) and natural killer (NK) cells. Alternatively or additionally, the ADCC activity of a molecule of interest can be assessed in vivo, such as in an animal model, such as described in Clynes et al. PNAS (USA) 95:652-656 (1998).

Complement-dependent cytotoxicity or CDC refers to the ability of a molecule to lyse a target in the presence of complement. The complement activation pathway is initiated by the binding of the first component of the complement system (C1q) to a molecule (e.g., an antibody) complexed with the antigen it recognizes. To assess complement activation, a CDC assay can be performed, such as that described by Gazzano-Santoro et al, J. Immunol. Methods 202:163 (1996).

Binding affinity refers to the strength of the total number of non-covalent interactions between a single binding site of a molecule (e.g., an antibody) and its binding partner (e.g., an antigen). Unless otherwise specified, as used herein, binding affinity refers to the affinity of self-binding, which reflects a 1:1 interaction between members of a binding pair (e.g., an antibody and an antigen). The affinity of a molecule X for its partner Y can generally be expressed by the dissociation constant (Kd). Affinity can be measured by conventional methods known in the art. Low-affinity antibodies generally bind antigen slowly and tend to dissociate readily, whereas high-affinity antibodies generally bind antigen more rapidly and tend to remain bound.

As used herein, the term Kd or Kd value refers to the dissociation constant determined by biolayer interferometry using an Octet QK384 instrument (Fortebio Inc., Menlo Park, CA) in kinetic mode. For example, mouse Fc sensors are loaded with mouse Fc-fused antigen and then loaded into wells containing antibodies to measure concentration-dependent rates (kon). Antibody off-rates (koff) are measured in the final step when the sensors are loaded into wells containing buffer only. Kd is the ratio koff/kon. (For more information, see Concepcion, J, et al., Comb Chem High Throughput Screen, 12(8), 791-800, 2009.)

The terms treatment, cure, and the like are used herein to mean achieving a desired pharmacological and/or physiological effect. The effect may be prophylactic in terms of completely or partially preventing a disease or its symptoms and/or may be therapeutic in terms of partially or completely curing a disease and/or an adverse event associated with the disease. As used herein, the term treatment encompasses any treatment of a disease in a mammal and includes: (a) preventing the onset of a disease in a subject that may be predisposed to the disease but has not yet been diagnosed with the disease; (b) suppressing the disease, i.e., stopping its progression; or (c) alleviating the disease, i.e., causing the disease to regress. The therapeutic agent may be administered before, during, or after the onset of a disease or injury. Treatment of an ongoing disease,

- 14 048216 in which the treatment stabilizes or reduces the patient's undesirable clinical symptoms is of particular interest. Such treatment is preferably carried out until complete loss of function in the affected tissues. Therapy in a subject can be carried out during the symptomatic stage of the disease, and in some cases after the symptomatic stage of the disease.

The term therapeutically effective amount means the amount of an active agent that is necessary to produce a therapeutic effect in a subject. For example, a therapeutically effective amount is an amount that causes, attenuates, or otherwise improves pathological symptoms, disease progression, or physiological conditions associated with the disease, or that increases resistance to a disorder.

The terms B-cell neoplasms or mature B-cell neoplasms in the context of the present invention include small lymphocytic lymphoma, B-cell prolymphocytic lymphoma, B-cell chronic lymphocytic leukemia, mantle cell lymphoma, Burkitt's lymphoma, follicular lymphoma, diffuse large B-cell lymphoma (DLBCL), multiple myeloma, lymphoplasmacytic lymphoma, splenic marginal zone lymphoma, plasma cell neoplasm such as plasma cell myeloma, plasmacytoma, monoclonal immunoglobulin deposition disease, heavy chain disease, MALT lymphoma, nodular marginal zone B-cell lymphoma, intravascular large B-cell lymphoma, primary effusion lymphoma, lymphomatoid granulomatosis, non-Hodgkin's lymphoma, Hodgkin's lymphoma, hairy cell leukemia, primary effusion lymphoma and AIDS-related non-Hodgkin's lymphoma.

The term characterized by CD22 expression broadly refers to any disease or disorder in which CD22 expression is associated with or involved in one or more pathological processes that are characteristic of that disease or disorder. Such disorders include, but are not limited to, B-cell neoplasms.

The terms subject, individual, and patient are used interchangeably herein to refer to a mammal that is being evaluated for treatment and/or that is being treated. In one embodiment, the mammal is a human. The terms subject, individual, and patient include, without limitation, individuals with a malignancy, individuals with autoimmune diseases, pathogenic infections, and the like. Subjects may be humans, but may also be other mammals, particularly those mammals that can be used as laboratory models of human diseases, such as a mouse, a rat, etc.

The term pharmaceutical formulation refers to a formulation that is in a form that provides the biological activity of the active ingredient to be effective and that does not contain any additional components that are unacceptably toxic to the subject to which the formulation is to be administered. Such formulations are sterile. Pharmaceutically acceptable excipients (carriers, adjuvants) are those that can reasonably be administered to a mammalian subject and provide an effective dose of the active ingredient used.

A sterile composition is aseptic or free from, or substantially free from, any living microorganisms and their spores. A frozen composition is a composition at a temperature below 0°C.

A stable composition is one in which the protein substantially completely retains its physical stability and/or chemical stability and/or biological activity during storage. Preferably, the composition substantially completely retains its physical or chemical stability as well as its biological activity. The storage period is generally selected based on the shelf life of the formulation. A variety of techniques for measuring protein stability are known in the art and are discussed, for example, in Peptide and Protein Drug Delivery, 247-301. Vincent Lee Ed., Marcel Dekker, Inc., New York, N.Y., Pubs. (1991) and Jones. A. Adv. Drug Delivery Rev. 10: 2990) (1993). Stability can be measured at a selected temperature for a selected period of time. Stability can be assessed qualitatively and/or quantitatively by a variety of means, including assessment of aggregate formation (e.g., using size exclusion chromatography, by measuring turbidity and/or by visual inspection); by assessing charge heterogeneity using cation exchange chromatography, isoelectric image focusing (icIEF), or capillary zone electrophoresis; amino-terminal or carboxy-terminal sequence analysis; mass spectrometric analysis; sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis to compare reduced and intact antibody; peptide map analysis (e.g., tryptic or LYS-C); assessment of biological activity or antigen-binding function of the antibody; etc. Instability may include one or more of: aggregation, deamidation (e.g., Asn deamidation), oxidation (e.g., Met oxidation), isomerization (e.g., Asp isomerization), truncation/hydrolysis/fragmentation (e.g., hinge region fragmentation), succinimide formation, unpaired cysteine(s), N-terminal extension, C-terminal processing, glycosylation differences, etc.

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II. Detailed description.

Antibodies to CD22.

Aspects of the invention include multispecific binding compounds that comprise an anti-CD22 binding domain. Provided herein is a family of closely related binding domain heavy chain-only antibodies that bind to human CD22. The antibodies of this family comprise a set of CDR sequences as defined herein and shown in Table 1 and are illustrated by the provided heavy chain variable region (VH) sequences of SEQ ID NOs: 24-84 shown in Table 2. The antibodies provided herein provide a number of advantages that favor their use as therapeutic agent(s) for clinical purposes. The antibodies include members with a range of binding affinities, allowing selection of a particular sequence with a desired binding affinity.

Table 1

Unique amino acid sequences of the CDRs of the heavy chain-only anti-CD22 antibody

SEQ aa CDR1 SEQ aa CDR2 SEQ aaCDR3 GDSISSGDYY (SEQ ID NO: 1) IYYSGVT (SEQ ID NO: 11) TREDSSNWRS (SEQ ID NO: 18) GDSISSGGYY (SEQ ID NO: 2) IYYSGAT (SEQ ID NO: 12) TRDDSSNWRS (SEQ ID NO: 19) GGSISSGDYY (SEQ ID NO: 3) IYYSGAT (SEQ ID NO: 13) TREDSSSSWRS (SEQ ID NO: 20) GGSISSSSYY (SEQ ID NO: 4) IYYTGST (SEQ ID NO: 14) AREDSSSWRS (SEQ ID NO: 21) GGSFSGYY (SEQ ID NO: 5) VYYTGAT (SEQ ID NO: 15) KRDDSSNWRS (SEQ ID NO: 22) GDSISSSSYY (SEQ ID NO: 6) IHYSGST (SEQ ID NO: 16) ARDDSSNWRS (SEQ ID NO: 23) GGSITSSSYY (SEQ ID NO: Ό IYYSGSA (SEQ ID NO: 17) GGSISSSSHY (SEQ ID NO: 8) GGSIISSSYY (SEQ ID NO: 9) GGSINDNSHY (SEQ ID NO: 10)

Table 2

Amino acid sequences of the variable domain of the heavy chain-only antibody to CD22

Clone ID SEQ_aa_FRl_FR4 SEQ ID NO: 335207 QLQLQESGPGLVKPSETLSLTCTVSGDSISSGDYYWGWIRQPP GKGLEWIGHI YYSGVTYYNPSLKSRVTISVDTSRNQFSLKLSS VT AADT A VYYCTRED S SNWRSRGQGTL VT VS S 24 335161 QLQLQESGPGLVKPSETLSLTCTVSGDSISSGDYYWGWIRQPP GKGLEWIGHI YYSGATYYNPSLENRVTISVDTSKNQFSLKLSS VT AADT A VYYCTRDD S SNWRSRGQGTL VT VS S 25 335254 QLQLQESGPGLVKPSETLSLTCTVSGDSISSGDYYWGWIRQPP GKGLEWIGHIYYSGVT YYNPSLKNRVTIS VDTSKNQF SLKLS S VTAADTAVYYCTREDSSSWRSRGQGTLVTVSS 26 335260 QLQLQESGPGLVKPSETLSLTCTVSGDSISSGDYYWGWIRQPP GKGLEWIGHI YYSGVTYYNPSLKNRVTISVDTSRNQFSLNLSS VTAADTAVYYCTREDSSSWRSRGQGTLVTVSS 27 335151 QLQLQESGPGLVKPSETLSLTCTVSGDSISSGDYYWGWIRQPP GKGLEWIGHI YYSGAT YYNPSLKNRVTIS VDTSRNQF SLNLS S VT AADT A VYYCTRDD S SNWRSRGQGTL VT VS S 28 335170 QLQLQESGPGLVKPSETLSLTCTVSGDSISSGDYYWGWIRQPP GKGLEWIGHI YYSGATYYNPSLKNRVTISVDTSRNQFSLKLSS VT AADT A VYYCTRDD S SNWRSRGQGTL VT VS S 29

- 16 048216

335176 QLQLQESGPGLVKPSETLSLTCTVSGDSISSGDYYWGWIRQPP GKGLEWIGHIYYSGATYYNPSLKNRVTISVDTSKNQFSLKLSS VT AADTAVYYCTRDD S SNWRSRGQGTL VT VS S 30 335181 QLQLQESGPGLVKPSETLSLTCTVSGDSISSGGYYWGWIRQPP GKGLEWIGHIYYSGATYYNPSLKNRVTISVDTSKNQFSLKLSS VT AADT AVYYCTRDD S SNWRSRGQGTL VT VS S 31 335244 QLQLQESGPGLVKPSETLSLTCTVSGDSISSGDYYWGWIRQPP GKGLEWIGHIYYSGATYYNPSLKNRVTISVDTSRNQFSLNLSS VTAADTAVYYCTREDSSSWRSRGQGTLVTVSS 32 335154 QLQLQESGPGLVKPSETLSLTCTVSGDSISSGDYYWGWIRQPP GKGLEWIGHIYYSGVT YYNPSLKNRVTIS VDT SKNQF SLKLS S VT AADT AVYYCTRDD S SNWRSRGQGTL VT VS S 33 335201 QLQLQESGPGLVKPSETLSLTCTVSGDSISSGDYYWGWIRQPP GKGLEWIGHIYYSGVTYYNPSLKNRVTISVDTSRNQFSLKLSS VT AADT AVYYCTRED S SNWRSRGQGTL VT VS S 34 335261 QLQLQESGPGLVKPSETLSLTCTVSGDSISSGDYYWGWIRQPP GKGLEWIGHIYYSGAT YYNPSLENRVTIS VDTSKNQF SLKLS S VTAADTAVYYCTREDSSSWRSRGQGTLVTVSS 35 335293 QLQLQESGPGLVKPSETLSLTCTVSGDSISSGDYYWGWIRQPP GKGLEWIGHIYYSGATYYNPSLKNRVTISVDTSRNQFSLKLSS VTAADTAVYYCTREDSSSWRSRGQGTLVTVSS 36 335203 QLQLQESGPGLVKPSETLSLTCTVSGDSISSGDYYWGWIRQPP GKGLEWIGHIYYSGVT YYNPSLKNRVTIS VDT SKNQF SLKLS S VT AADT AVYYCTRED S SNWRSRGQGTL VT VS S 37 335185 QLQLQESGPGLVKPSETLSLTCTVSGDSISSGDYYWGWIRQPP GKGLEWIGHIYYSGATYYNPSLKNRVTISVDTSRNQFSLNLSS VT AADT AVYYCTRED S SNWRSRGQGTL VT VS S 38 335206 QLQLQESGPGLVKPSETLSLTCTVSGDSISSGDYYWGWIRQPP GKGLEWIGHIYYSGAT YYNPSLKNRVTIS VDT SKNQF SLKLS S VT AADT AVYYCTRED S SNWRSRGQGTL VT VS S 39 335245 QLQLQESGPGLVKPSETLSLTCTVSGDSISSGDYYWGWIRQPP GKGLEWIGHIYYSGATYYNPSLKNRVTISVDTSKNQFSLKLSS VTAADTAVYYCTREDSSSWRSRGQGTLVTVSS 40 335218 QLQLQESGPGLVKPSETLSLTCTVSGDSISSGDYYWGWIRQPP GKGLEWIGHIYYSGATYYNPSLKNRVTISVDTSRNQFSLKLSS VT AADT AVYYCTRED S SNWRSRGQGTL VT VS S 41 335160 QLQLQESGPGLVKPSETLSLTCTVSGDSISSGDYYWGWIRQPP GKGLEWIGSIYYSGAT YYNPSLKNRVTIS VDTSKNQF SLKLS S VT AADT AVYYCTRDD S SNWRSRGQGTL VT VS S 42 335158 QLQLQESGPGLVKPSETLSLTCTVSGGSISSGDYYWGWIRQPP GKGLEWIGHIYYSGAT YYNPSLKNRVTIS VDT SRNQF SLKLS S VT AADT AVYYCTRDD S SNWRSRGQGTL VT VS S 43 324508 QLQLQESGPGLVKPSETLSLTCTVSGGSISSGDYYWGWIRQPP GKGLEWIGHIYYSGATYYNPSLENRVTISVDTSKNQFSLKLSS VT AADT AVYYCTRDD S SNWRSRGQGTL VT VS S 44 335307 QLQLQESGPGLVKPSETLSLTCTVSGDSISSGDYYWGWIRQPP GKGLEWIGSIYYSGSTYYNPSLKSRVTISVDTSKNQFSLKLSS VTAADTAVYYCTREDSSSWRSRGQGTLVTVSS 45

- 17 048216

335301 QLQLQESGPGLVKPSETLSLTCTVSGDSISSGDYYWGWIRQPP GKGLEWIGNIYYSGAT YYNPSLKNRVTIS VDTSKNQF SLKLS S VTAADTAVYYCTREDSSSWRSRGQGTLVTVSS 46 335323 QLQLQESGPGLVKPSETLSLTCTVSGDSISSGGYYWGWIRQPP GKGLEWIGSIYYSGSTYYNPSLKNRVTISVDTSKNQFSLKLSS VTAADTAVYYCTREDSSSWRSRGQGTLVTVSS 47 335271 QLQLQESGPGLVKPSETLSLTCTVSGDSISSGDYYWGWIRHPP GKGLEWIGHIYYSGAT YYNPSLKNRVTIS VDTSKNQF SLKLS S VTAADTAVYYCTREDSSSWRSRGQGTLVTVSS 48 335234 QLQLQESGPGLVKPSETLSLTCTVSGDSISSGDYYWGWIRQPP GKGLEWIGNIYYSGAT YYNPSLKNRVTIS VDTSKNQF SLKLS S VT AADT AVYYCTRED S SNWRSRGQGTL VT VS S 49 335182 QLQLQESGPGLVKPSETLSLTCTVSGDSISSGDYYWGWIRQPP GKGLEWIGNIYYSGAT YYNPSLKNRVTIS VDTSKNQF SLKLS S VT AADT AVYYCTRDD S SNWRSRGQGTL VT VS S 50 335186 QLQLQESGPGLVKPSETLSLTCTVSGGSISSSSYYWGWIRQPP GKGLEWIGSIYYSGSTYYNPSLKSRVTISVDTSKNQFSLKLSS VT AADT AVYYCTRDD S SNWRSRGQGTL VT VS S 51 335233 QLQLQESGPGLVKPSETLSLTCTVSGDSISSGDYYWGWIRQPP GKGLEWIGSIYYSGSTYYNPSLKSRVTISVDTSKNQFSLKLSS VT AADT AVYYCTRED S SNWRSRGQGTL VT VS S 52 335224 QLQLQESGPGLVKPSETLSLTCTVSGDSISSGDYYWGWIRQPP GKGLEWIGSIYYTGSTYYNPSLKSRVTISVDTSKNQFSLKLSS VT AADT AVYYCTRED S SNWRSRGQGTL VT VS S 53 335210 QLQLQESGPGLVKPSETLSLTCTVSGGSISSGDYYWGWIRQPP GKGLEWIGHIYYSGATYYNPSLKNRVTISVDTSRNQFSLKLSS VT AADT AVYYCTRED S SNWRSRGQGTL VT VS S 54 335311 QLQLQESGPGLVKPSETLSLTCAVYGGSFSGYYWGWIRQPPG KGLEWIGHIYYSGATYYNPSLKNRVTISVDTSRNQFSLNLSSV TAADTAVYYCTREDSSSWRSRGQGTLVTVSS 55 335159 QLQLQESGPGLVKPSETLSLTCTVSGGSISSSSYYWGWIRQPP GKGLEWIGHIYYSGVT YYNPSLKNRVTIS VDTSKNQF SLKLS S VT AADT AAYYCTRDD S SNWRSRGQGTL VT VS S 56 335188 QLQLQESGPGLVKPSETLSLTCTVSGGSISSSSYYWGWIRQPP GKGLEWIGHIYYSGVT YYNPSLKNRVTIS VDTSKNQF SLKLS S VT AADT AVYYCTRDD S SNWRSRGQGTL VT VS S 57 335274 QLQLQESGPGLVKPSETLSLTCTVSGGSISSSSYYWGWIRQPP GKGLEWIGSIYYSGSTYYNPSLKSRVTISVDTSRNQFSLNLSSV TAADTAVYYCTREDSSSWRSRGQGTLVTVSS 58 335226 QLQLQESGPGLVKPSETLSLTCTVSGDSISSSSYYWGWIRQPP GKGLEWIGSIYYSGSTYYNPSLKSRVTISVDTSKNQFSLKLSS VT AADT AVYYCTRED S SNWRSRGQGTL VT VS S 59 335333 QLQLQESGPGLVKPSETLSLTCTVSGGSISSSSYYWGWIRQPP GKGLEWIGSIYYSGSTYYNPSLKSRVTISVDTSKNQFSLKLSS VT AADT AVYYC ARED S S S WRSRGQGTL VT VS S 60 335283 QLQLQESGPGLVKPSETLSLTCTVSGGSISSSSYYWGWIRQPP GKGLEWIGSIYYSGAT YYNPSLKNRVTIS VDTSKNQF SLKLS S VTAADTAVYYCTREDSSSWRSRGQGTLVTVSS 61

- 18 048216

335297 QLQLQESGPGLVKPSETLSLTCTVSGGSISSSSYYWGWIRQPP GKGLEWIGHIYYSGATYYNPSLKNRVTISVDTSRNQFSLNLSS VTAADTAVYYCTREDSSSWRSRGQGTLVTVSS 62 335273 QLQLQESGPGLVKPSETLSLTCTVSGGSISSSSYYWGWIRQPP GKGLEWIGSIYYSGSTYYNPSLKSRVTISVDTSKNQFSLKLSS VTAADTAVYYCTREDSSSWRSRGQGTLVTVSS 63 335187 QLQLQESGPGLVKPSETLSLTCTVSGGSISSSSYYWGWIRQPP GKGLEWIGHIYYSGAT YYNPSLKNRVTIS VDTSKNQF SLKLS S VT AADT AVYYCTRDD S SNWRSRGQGTL VT VS S 64 335295 QLQLQESGPGLVKPSETLSLTCTVSGGSISSSSYYWGWIRQPP GKGLEWIGSIYYSGATYYNPSLKNRVTISVDTSRNQFSLNLSS VTAADTAVYYCTREDSSSWRSRGQGTLVTVSS 65 335220 QLQLQESGPGLVKPSETLSLTCTVSGGSISSSSYYWGWIRQPP GKGLEWIGSIYYSGSTYYNPSLKNRVTISVDTSKNQFSLKLSS VT AADT AVYYCTRED S SNWRSRGQGTL VTVS S 66 335173 QLQLQESGPGLVKPSETLSLTCTVSGGSITSSSYYWGWIRQPP GKGLEWIGSIYYSGSTYYNPSLKSRVTISVDTSKNQFSLKLSS VT AADT AVYYCTRDD S SNWRSRGQGTL VTVS S 67 335219 QLQLQESGPGLVKPSETLSLTCTVSGGSISSSSYYWGWIRQPP GKGLEWIGSIYYSGVT YYNPSLKNRVTIS VDTSKNQF SLKLS S VT AADT AVYYCTRED S SNWRSRGQGTL VTVS S 68 335236 QLQLQESGPGLVKPSETLSLTCTVSGGSISSSSYYWGWIRQPP GKGLEWIGSIYYSGSTYYNPSLKSRVTISVDTSKNQFSLKLSS VT AADT AVYYCTRED S SNWRSRGQGTL VTVS S 69 335266 QLQLQESGPGLVRPSETLSLTCTVSGGSISSSSYYWGWIRQPP GKGLEWIGSIYYSGSTYYNPSLKSRVTISVDTSKNQFSLKLSS VTAADTAVYYCTREDSSSWRSRGQGTLVTVSS 70 335208 QLQLQESGPGLVKPSETLSLTCTVSGGSISSSSYYWGWIRQPP GKGLEWIGSIYYSGATYYNPSLKNRVTISVDTSRNQFSLNLSS VT AADT AVYYCTRED S SNWRSRGQGTL VTVS S 71 335195 QLQLQESGPGLVKPSETLSLTCTVSGGSISSSSYYWGWIRQPP GKGLEWIGSIYYSGATYYNPSLKNRVTISVDTSRNQFSLNLSS VT AADT AMYYCTREDS SNWRSRGQGTL VTVS S 72 335285 QLQLQESGPGLVKPSETLSLTCTVSGGSISSSSYYWGWIRQPP GKGLEWIGSIYYSGVT YYNPSLKNRVTIS VDTSKNQF SLKLS S VTAADTAVYYCTREDSSSWRSRGQGTLVTVSS 73 335150 QLQLQESGPGLVKPSETLSLTCTVSGDSISSGDYYWGWIRQSP EKGLEWIGHIYYSGVTYYNPSLKNRVTISVDTSKNQFSLKLSS VT AADT AVYYCKRDD S SNWRSRGQGTL VTVS S 74 335316 QLQLQESGPGLVKPSETLSLTCTVSGGSISSSSYYWGWIRQPP GKGLEWIGHIYYSGAT YYNPSLKNRVTIS VDTSKNQF SLKLS S VTAADTAVYYCTREDSSSWRSRGQGTLVTVSS 75 335189 QLQLQESGPGLVKPSETLSLTCTVSGGSISSSSYYWGWIRQPP GKGLEWIGSVYYTGATYYNPSLKNRVTISVDTSKNQFSLKLS SVTAADTAVYYCTRDDSSNWRSRGQGTLVTVSS 76 335179 QLQLQESGPGLVKPSETLSLTCTVSGGSISSSSYYWGWFRHPP GKGLDWIGSIHYSGSTYYNPSLKSRVTISVDTSRNQFSLNLSS VT AADT AVYYCTRDD S SNWRSRGQGTL VTVS S 77 335230 QLQLQESDPGLVKPSETL SLTCT VSGGSIS S S SHYWGWIRQPP GKGLEWIGHIYYSGATYYNPSLKNRVTISVDTSRNQFSLNLSS VT AADT AVYYCTRED S SNWRSRGQGTL VTVS S 78 335166 QLQLQESGPGLVKPSETLSLTCTVSGGSISSSSYYWGWIRQPP GKGLEWIGSIYYSGSTYYNPSLKNRVTISVDTSRNQFSLNLSS VT AADT AVYYCTRDD S SNWRSRGQGTL VTVS S 79 335242 QLQLQESGPGLVKPSETLSLTCTVSGGSISSSSYYWGWIRQPP GKGLEWIGHIYYSGATYYNPSLKNRVTISVDTSRNQFSLNLSS VT AADT AVYYCTRED S SNWRSRGQGTL VTVS S 80 335162 QLQLQESGPGLVKPSETLSLTCTVSGGSIISSSYYWGWIRQPPG KGLEWIGSIYYSGSAYYHPSLKSRVTISIDTSKNQFSLKLSSVT AADT AVYYC ARDD S SNWRSRGQGTL VTVS S 81 335171 QLQLQESGPGLVKPSETLSLTCTVSGGSISSSSYYWGWIRQPP GKGLEWIGSIYYSGATYYNPSLKNRVTISVDTSRNQFSLNLSS VT AADT AVYYCTRDD S SNWRSRGQGTL VTVS S 82 335232 QLQLQESGPGLVKPSETLSLTCTVSGDSISSGDYYWGWIRQPP GKGLEWIGHIYYSGATYYNPSLKNRVTISVDTSRNQSSLNLSS VT AADT AVYYCTRED S SNWRSRGQGTL VTVS S 83 335263 QLQLQESGPGLVKPSETLSLTCTVSGGSINDNSHYWGWIRQPP GKGLEWIGHIYYSGATYYNPSLKNRVTISVDTSRNQFSLNLSS VTAADTAVYYCTREDSSSWRSRGQGTLVTVSS 84

A suitable antibody may be selected from those provided herein for development and therapeutic or other use, including, but not limited to, use as a bis- 19 048216 specific, such as shown in Fig. 14A, or trispecific antibody or part of a CAR-T structure (e.g., as shown in Fig. 14B). Fig. 14A provides an illustration of a non-limiting example of a multispecific anti-CD3 x CD22 antibody, wherein the anti-CD22 domain is monovalent and monospecific. In some embodiments, the anti-CD3 domain comprises a CH1 domain and is paired with a light chain, while the anti-CD22 domain(s) are derived from heavy chain-only antibodies and lack a CH1 domain and do not interact with a light chain. In some embodiments, the two heavy chains are separated using, for example, knob-in-hole technology.

Referring to the antibodies depicted in Fig. 15, Fig. 15A shows an anti-CD3 x CD22 bispecific antibody in which the anti-CD22 binding arm is monovalent and monospecific and the antigen-binding domain of the anti-CD22 arm is in a single configuration, meaning that only one antigen-binding domain is present. Fig. 15B shows an anti-CD3 x CD22 bispecific antibody in which the anti-CD22 binding arm is bivalent and monospecific and the antigen-binding domain of the anti-CD22 arm is in a tandem configuration, meaning that there are two identical antigen-binding domains arranged in tandem. Fig. 15C shows an anti-CD3 x CD22 bispecific antibody in which the anti-CD22 binding arm is bivalent and biparatopic and the antigen-binding domains of the anti-CD22 arm are in a tandem configuration.

Determining the affinity for a candidate protein can be accomplished using methods known in the art, such as Biacore measurements. The antibody family members can have an affinity for CD22 with a Kd of from about 10 ^-6 to about 10 ^-11 , including, but not limited to: from about 10 ^-6 to about 10 ^-10 ; from about 10 ^-6 to about 10 ^-9 ; from about 10 ^-6 to about 10 ^-8 ; from about 10 ^-8 to about 10'¹¹; from about 10 ⁸ to about 10 ¹⁰ ; from about 10 ^-8 to about 10 ^-9 ; from about 10 ^-9 to about 10 ^-11 ; from about 10 ^-9 to about 10 ^-10 ; or any value within these ranges. Affinity selection can be supported by biological assessment for modulation, such as blockade, of CD22 biological activity, including in vitro assays, preclinical models and clinical trials, as well as assessment of potential toxicity.

The antibody family members described herein are not cross-reactive with cynomolgus monkey CD22 protein, but can be engineered to be cross-reactive with cynomolgus monkey CD22 protein or CD22 proteins from other animal species, if desired.

The family of CD22-specific antibodies herein comprises a VH domain comprising CDR1, CDR2, and CDR3 sequences within a human VH framework. The CDR sequences can be located, for example, in the region around amino acid residues 26-35; 53-59; and 98-117 for CDR1, CDR2, and CDR3, respectively, of the provided exemplary variable region sequences set forth in SEQ ID NOs: 24-84. One skilled in the art will appreciate that the CDR sequences can be located at other positions if a different framework sequence is selected, although the order of the sequences will generally remain unchanged.

The CDR1, CDR2, and CDR3 sequences of the anti-CD22 antibodies of the present invention may be encompassed by the following structural formulas, wherein X is a variable amino acid, which may be a variable amino acid as defined below.

CDR1

G Xi SI Xg Xz X ₄ X ₅ Xb Y (SEQ ID NO: 104) where X1 is D or G;

X2 represents S, T, I or N;

X ₃ represents S or D;

X4 represents G, S or N;

X5 is D, G, or S; and

X ₆ represents Y or H.

CDR2

X? x ₈ Y X9 G Xu Xn (SEQ ID NO: 105) where X ₇ is I or V;

X ₈ represents Y or H;

X ₉ represents S or T;

X ₁₀ represents A, V, or S; and

X11 represents T or A.

CDR3

X12 R X13 DSS Xi WRS (SEQ ID NO: 106) where X ₁₂ is T, A or K;

X ₁₃ represents D or E; and

X ₁₄ represents N or S.

Typical sequences of CDR1, CDR2, and CDR3 are shown in Tables 1 and 3.

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Table 3 Amino acid sequences of CDR1, CDR2, and CDR3 of the heavy chain-only antibody to CD22

Clone ID SEQaaCDRl SEQ_aa_CDR2 SEQ_aaCDR3 335207 GDSISSGDYY (SEQ IDNO: 1) IYYSGVT (SEQ ID NO: 11) TREDSSNWRS (SEQ ID NO: 18) 335161 GDSISSGDYY (SEQ IDNO: 1) IYYSGAT (SEQ ID NO: 12) TRDDSSNWRS (SEQ ID NO: 19) 335254 GDSISSGDYY (SEQ IDNO: 1) IYYSGVT (SEQ ID NO: 11) TREDSSSSWRS (SEQ ID NO: 20) 335260 GDSISSGDYY (SEQ IDNO: 1) IYYSGVT (SEQ ID NO: 11) TREDSSSSWRS (SEQ ID NO: 20) 335151 GDSISSGDYY (SEQ IDNO: 1) IYYSGAT (SEQ ID NO: 12) TRDDSSNWRS (SEQ ID NO: 19) 335170 GDSISSGDYY (SEQ IDNO: 1) IYYSGAT (SEQ ID NO: 12) TRDDSSNWRS (SEQ ID NO: 19) 335176 GDSISSGDYY (SEQ IDNO: 1) IYYSGAT (SEQ ID NO: 12) TRDDSSNWRS (SEQ ID NO: 19) 335181 GDSISSGGYY (SEQ ID NO: 2) IYYSGAT (SEQ ID NO: 12) TRDDSSNWRS (SEQ ID NO: 19) 335244 GDSISSGDYY (SEQ IDNO: 1) IYYSGAT (SEQ ID NO: 12) TREDSSSSWRS (SEQ ID NO: 20) 335154 GDSISSGDYY (SEQ IDNO: 1) IYYSGVT (SEQ ID NO: 11) TRDDSSNWRS (SEQ ID NO: 19)

- 21 048216

335201 GDSISSGDYY (SEQ ID NO: 1) IYYSGVT (SEQ ID NO: 11) TREDSSNWRS (SEQ ID NO: 18) 335261 GDSISSGDYY (SEQ ID NO: 1) IYYSGAT (SEQ ID NO: 12) TREDSSSSWRS (SEQ ID NO: 20) 335293 GDSISSGDYY (SEQ ID NO: 1) IYYSGAT (SEQ ID NO: 12) TREDSSSSWRS (SEQ ID NO: 20) 335203 GDSISSGDYY (SEQ ID NO: 1) IYYSGVT (SEQ ID NO: 11) TREDSSNWRS (SEQ ID NO: 18) 335185 GDSISSGDYY (SEQ ID NO: 1) IYYSGAT (SEQ ID NO: 12) TREDSSNWRS (SEQ ID NO: 18) 335206 GDSISSGDYY (SEQ ID NO: 1) IYYSGAT (SEQ ID NO: 12) TREDSSNWRS (SEQ ID NO: 18) 335245 GDSISSGDYY (SEQ ID NO: 1) IYYSGAT (SEQ ID NO: 12) TREDSSSSWRS (SEQ ID NO: 20) 335218 GDSISSGDYY (SEQ ID NO: 1) IYYSGAT (SEQ ID NO: 12) TREDSSNWRS (SEQ ID NO: 18) 335160 GDSISSGDYY (SEQ ID NO: 1) IYYSGAT (SEQ ID NO: 12) TRDDSSNWRS (SEQ ID NO: 19) 335158 GGSISSGDYY (SEQ ID NO: 3) IYYSGAT (SEQ ID NO: 12) TRDDSSNWRS (SEQ ID NO: 19) 324508 GGSISSGDYY (SEQ ID NO: 3) IYYSGAT (SEQ ID NO: 12) TRDDSSNWRS (SEQ ID NO: 19) 335307 GDSISSGDYY (SEQ ID NO: 1) IYYSGST (SEQ ID NO: 13) TREDSSSSWRS (SEQ ID NO: 20) 335301 GDSISSGDYY (SEQ ID NO: 1) IYYSGAT (SEQ ID NO: 12) TREDSSSSWRS (SEQ ID NO: 20) 335323 GDSISSGGYY (SEQ ID NO: 2) IYYSGST (SEQ ID NO: 13) TREDSSSSWRS (SEQ ID NO: 20) 335271 GDSISSGDYY (SEQ ID NO: 1) IYYSGAT (SEQ ID NO: 12) TREDSSSSWRS (SEQ ID NO: 20) 335234 GDSISSGDYY (SEQ ID NO: 1) IYYSGAT (SEQ ID NO: 12) TREDSSNWRS (SEQ ID NO: 18) 335182 GDSISSGDYY (SEQ ID NO: 1) IYYSGAT (SEQ ID NO: 12) TRDDSSNWRS (SEQ ID NO: 19) 335186 GGSISSSSYY (SEQ ID NO: 4) IYYSGST (SEQ ID NO: 13) TRDDSSNWRS (SEQ ID NO: 19) 335233 GDSISSGDYY (SEQ ID NO: 1) IYYSGST (SEQ ID NO: 13) TREDSSNWRS (SEQ ID NO: 18) 335224 GDSISSGDYY (SEQ ID NO: 1) IYYTGST (SEQ ID NO: 14) TREDSSNWRS (SEQ ID NO: 18) 335210 GGSISSGDYY (SEQ ID NO: 3) IYYSGAT (SEQ ID NO: 12) TREDSSNWRS (SEQ ID NO: 18) 335311 GGSFSGYY (SEQ ID NO: 5) IYYSGAT (SEQ ID NO: 12) TREDSSSSWRS (SEQ ID NO: 20) 335159 GGSISSSSYY (SEQ ID NO: 4) IYYSGVT (SEQ ID NO: 11) TRDDSSNWRS (SEQ ID NO: 19) 335188 GGSISSSSYY (SEQ ID NO: 4) IYYSGVT (SEQ ID NO: 11) TRDDSSNWRS (SEQ ID NO: 19)

- 22 048216

335274 GGSISSSSYY (SEQ ID NO: 4) IYYSGST (SEQ ID NO: 13) TREDSSSSWRS (SEQ ID NO: 20) 335226 GDSISSSSYY (SEQ ID NO: 6) IYYSGST (SEQ ID NO: 13) TREDSSNWRS (SEQ ID NO: 18) 335333 GGSISSSSYY (SEQ ID NO: 4) IYYSGST (SEQ ID NO: 13) AREDSSSWRS (SEQ ID NO: 21) 335283 GGSISSSSYY (SEQ ID NO: 4) IYYSGAT (SEQ ID NO: 12) TREDSSSSWRS (SEQ ID NO: 20) 335297 GGSISSSSYY (SEQ ID NO: 4) IYYSGAT (SEQ ID NO: 12) TREDSSSSWRS (SEQ ID NO: 20) 335273 GGSISSSSYY (SEQ ID NO: 4) IYYSGST (SEQ ID NO: 13) TREDSSSSWRS (SEQ ID NO: 20) 335187 GGSISSSSYY (SEQ ID NO: 4) IYYSGAT (SEQ ID NO: 12) TRDDSSNWRS (SEQ ID NO: 19) 335295 GGSISSSSYY (SEQ ID NO: 4) IYYSGAT (SEQ ID NO: 12) TREDSSSSWRS (SEQ ID NO: 20) 335220 GGSISSSSYY (SEQ ID NO: 4) IYYSGST (SEQ ID NO: 13) TREDSSNWRS (SEQ ID NO: 18) 335173 GGSITSSSYY (SEQ ID NO: 7) IYYSGST (SEQ ID NO: 13) TRDDSSNWRS (SEQ ID NO: 19) 335219 GGSISSSSYY (SEQ ID NO: 4) IYYSGVT (SEQ ID NO: 11) TREDSSNWRS (SEQ ID NO: 18) 335236 GGSISSSSYY (SEQ ID NO: 4) IYYSGST (SEQ ID NO: 13) TREDSSNWRS (SEQ ID NO: 18) 335266 GGSISSSSYY (SEQ ID NO: 4) IYYSGST (SEQ ID NO: 13) TREDSSSSWRS (SEQ ID NO: 20) 335208 GGSISSSSYY (SEQ ID NO: 4) IYYSGAT (SEQ ID NO: 12) TREDSSNWRS (SEQ ID NO: 18) 335195 GGSISSSSYY (SEQ ID NO: 4) IYYSGAT (SEQ ID NO: 12) TREDSSNWRS (SEQ ID NO: 18) 335285 GGSISSSSYY (SEQ ID NO: 4) IYYSGVT (SEQ ID NO: 11) TREDSSSSWRS (SEQ ID NO: 20) 335150 GDSISSGDYY (SEQ ID NO: 1) IYYSGVT (SEQ ID NO: 11) KRDDSSNWRS (SEQ ID NO: 22) 335316 GGSISSSSYY (SEQ ID NO: 4) IYYSGAT (SEQ ID NO: 12) TREDSSSSWRS (SEQ ID NO: 20) 335189 GGSISSSSYY (SEQ ID NO: 4) VYYTGAT (SEQ ID NO: 15) TRDDSSNWRS (SEQ ID NO: 19) 335179 GGSISSSSYY (SEQ ID NO: 4) H4YSGST (SEQ ID NO: 16) TRDDSSNWRS (SEQ ID NO: 19) 335230 GGSISSSSHY (SEQ ID NO: 8) IYYSGAT (SEQ ID NO: 12) TREDSSNWRS (SEQ ID NO: 18) 335166 GGSISSSSYY (SEQ ID NO: 4) IYYSGST (SEQ ID NO: 13) TRDDSSNWRS (SEQ ID NO: 19) 335242 GGSISSSSYY (SEQ ID NO: 4) IYYSGAT (SEQ ID NO: 12) TREDSSNWRS (SEQ ID NO: 18) 335162 GGSIISSSYY (SEQ Ш NO: 9) IYYSGSA (SEQ ID NO: 17) ARDDSSNWRS (SEQ ID NO: 23) 335171 GGSISSSSYY (SEQ Ш NO: 4) IYYSGAT (SEQ ID NO: 12) TRDDSSNWRS (SEQ ID NO: 19) 335232 GDSISSGDYY (SEQ ID NO: 1) IYYSGAT (SEQ ID NO: 12) TREDSSNWRS (SEQ ID NO: 18) 335263 GGSINDNSHY (SEQ ID NO: 10) IYYSGAT (SEQ ID NO: 12) TREDSSSSWRS (SEQ ID NO: 20)

In some embodiments, a heavy chain-only anti-CD22 antibody of the present invention comprises a CDR1 sequence of any one of SEQ ID NOs: 1-10. In a particular embodiment, the CDR1 sequence is SEQ ID NO: 1.

In some embodiments, a heavy chain-only anti-CD22 antibody of the present invention comprises a CDR2 sequence of any one of SEQ ID NOs: 11-17. In a particular embodiment, the CDR2 sequence is SEQ ID NO: 11.

In some embodiments, a heavy chain-only anti-CD22 antibody of the present invention comprises a CDR3 sequence of any one of SEQ ID NOs: 18-23. In a particular embodiment, the CDR2 sequence is SEQ ID NO: 18.

In a further embodiment, a heavy chain-only anti-CD22 antibody of the present invention comprises a CDR1 sequence of SEQ ID NO: 1; a CDR2 sequence of SEQ ID NO: 11; and a CDR3 sequence of SEQ ID NO: 18.

In further embodiments, a heavy chain-only anti-CD22 antibody of the present invention comprises any of the amino acid sequences of a variable

- 23 048216 heavy chain regions of SEQ ID NO: 24-84 (Table 2).

In another embodiment, a heavy chain-only anti-CD22 antibody of the present invention comprises the heavy chain variable region of SEQ ID NO: 24.

In some embodiments, a CDR sequence in an anti-CD22 heavy chain only antibody of the invention comprises one or two amino acid substitutions relative to a CDR1, CDR2, and/or CDR3 sequence or a set of CDR1, CDR2, and CDR3 sequences in any of SEQ ID NOs: 1-23 (FIG. 1). In some embodiments, said amino acid substitution(s) are one or two amino acid positions 4-6 of CDR1 and/or one or two amino acid positions 2, 4-7 of CDR2 and/or one or two amino acid positions 5 and 12 of CDR3 relative to the formulas provided above. In some embodiments, heavy chain-only anti-CD22 antibodies may comprise a heavy chain variable region sequence with at least about 85% identity, at least 90% identity, at least 95% identity, at least 98% identity, or at least 99% identity to any one of the heavy chain variable region sequences of SEQ ID NOs: 24-84 (as set forth in Table 2).

In some embodiments, bispecific or multispecific antibodies are provided, which may have any of the configurations discussed herein, including, but not limited to, a bispecific three-chain antibody-like molecule. In some embodiments, a multispecific antibody may comprise at least one heavy chain variable region having binding specificity for CD22. In some embodiments, a multispecific antibody may comprise a heavy chain variable region comprising at least two antigen-binding domains, wherein each of the antigen-binding domains has binding specificity for CD22. In some embodiments, a multispecific antibody may comprise a heavy chain/light chain pair that has binding specificity for a first antigen (e.g., CD3) and a heavy chain from an antibody comprising only heavy chains. In some embodiments, the heavy chain of the heavy chain-only antibody comprises an Fc portion comprising CH2 and/or CH3 and/or CH4 domains, in the absence of a CH1 domain. In one particular embodiment, the bispecific antibody comprises a heavy chain/light chain pair that has binding specificity for an antigen on an effector cell (e.g., a CD3 protein on a T cell), and a heavy chain from a heavy chain-only antibody comprising an antigen-binding domain that has binding specificity for CD22.

In some embodiments, the multispecific antibody comprises a VH CD3 binding domain that is paired with a light chain variable domain. In certain embodiments, the light chain is a fixed light chain. In some embodiments, the VH CD3 binding domain comprises a CDR1 sequence of SEQ ID NO: 85, a CDR2 sequence of SEQ ID NO: 86, and a CDR3 sequence of SEQ ID NO: 87 in a human VH framework. In some embodiments, the fixed light chain comprises a CDR1 sequence of SEQ ID NO: 88, a CDR2 sequence of SEQ ID NO: 89, and a CDR3 sequence of SEQ ID NO: 90 in a human VL framework. Together, the VH CD3 binding domain and the light chain variable domain have a binding affinity for CD3. In some embodiments, the CD3 binding domain of the VH comprises a heavy chain variable region sequence of SEQ ID NO: 91. In some embodiments, the CD3 binding domain of the VH comprises a sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, or at least about 99% identity to the heavy chain variable region sequence of SEQ ID NO: 91. In some embodiments, the fixed light chain comprises a light chain variable region sequence of SEQ ID NO: 92. In some embodiments, the fixed light chain comprises a sequence having at least about 80%, at least about 85%, at least about 90%, at least about 95%, or at least about 99% identity to the heavy chain variable region sequence of SEQ ID NO: 92.

Multispecific antibodies comprising the CD3-binding VH domain and the variable light chain domain described above have useful properties, for example, as described in PCT Published Publication No. WO2018/052503, the disclosure of which is incorporated herein by reference in its entirety. Any of the multispecific antibodies and antigen-binding domains having binding affinity for CD22 described herein can be combined with the CD3-binding domains and fixed light chain domains described herein to create multispecific antibodies having binding affinity for one or more epitopes of CD22, as well as for CD3.

- 24 048216

Table 4

Amino acid sequences of CDR1, CDR2, CDR3 of heavy and light chains of CD3

SEQ aa CDR1 SEQ aa CDR2 SEQ aa CDR3 Heavy chain GFTFHNYA ISWNSGSI AKDSRGYGDYSLGGAY (SEQIDNO: 85) (SEQIDNO: 86) (SEQ ID NO: 87) Light chain QSVSSN (SEQIDNO: 88) GAS (SEQIDNO: 89) QQYNNWPWT (SEQ ID NO: 90)

Table 5

Amino acid sequences of the variable regions of the heavy and light chains of CD3

VH EVQLVESGGGLVQPGRSLRLSCAASGFTFHNYAMHWVRQAPGKGLEWVS GISWNSGSIGYADSVKGRFTISRDNAKNSLYLQMNSLRAEDTALYYCAKDS RGYGDYSLGGAYWGQGTLVTVSSDYRLGGAYWGQGTLVTVSS (SEQ ID NO: 91) VL EIVMTQSPATLSVSPGERATLLSCRASQSVSSNLAWYQQKPGQAPRLLIYG ASTRATGIPARFSGSGSGTEFTLTISSLQSEDFAVYYCQQYNNWPWTFGQ GTKVEIK (SEQ ID NO: 92)

Table 6

Sequences of the Fc regions of human IgG1 and IgG4

Human IgGl (UniProt P01857) No. ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVS WNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQT YICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGG PSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNW YVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGK EYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDE LTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPV LDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYT QKSLSLSPGK (SEQ ID NO: 93) Human IgG4 (UniProt P01861) No. ASTKGPSVFP LAPCSRSTSESTAALGCLVKDYFPEPVTVS WNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKT YTCNVDHKPSNTKVDKRVESKYGPPCPSCPAPEFLGGPSV FLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVD GVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYK CKVSNKGLP S SIEKTISKAKGQPREPQ VYTLPP SQEEMTK NQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDS DGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKS LSLSLGK (SEQ ID NO: 94) Human IgGl with silent mutation (Fc region) ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSG ALTSGVHTFP AVLQS SGL YSLS S VVT VPS S SLGTQT YICNVNHKP SNTKVDKKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDT LMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPR EEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKT ISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAV EWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNV FSCSVMHEALHNHYTQKSLSLSPGK (SEQ ID NO: 95) Human IgG4 with silent mutation (Fc region) ASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSG ALTSGVHTFP AVLQS SGL YSLS SWT VPS S SLGTKT YTCNVDHK PSNTKVDKRVESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMI SC S VMHEALHNHYTQKSLSLSLGK (SEQ ID NO: 96)

- 25 048216

Table 7

Additional sequences

Sequence of the constant region of the light chain to CD3 (light kappa chain) EIVMTQSPATLSVSPGERATLSCRASQSVSSNLAWYQQKPGQAP RLLIYGASTRATGIPARFSGSGSGTEFTLTISSLQSEDFAVYYCQQ YNNWPWTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVC LLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSS TLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 97) CD3 heavy chain sequence (with wild-type IgGl Fc) EVQLVESGGGLVQPGRSLRLSCAASGFTFHNYAMHWVRQAPG KGLEWVSGISWNSGSIGYADSVKGRFTISRDNAKNSLYLQMNS LRAEDTALYYCAKDSRGYGDYSLGGAYWGQGTLVTVSSASTK GPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTS GVHTFP AVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNT KVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMIS RTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQY NSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKA KGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWES NGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCS VMHEALHNHYTQKSLSLSPGK (SEQ ID NO: 98) CD3 heavy chain constant region sequence (with IgGl Fc silenced) EVQLVESGGGLVQPGRSLRLSCAASGFTFHNYAMHWVRQAPG KGLEWVSGISWNSGSIGYADSVKGRFTISRDNAKNSLYLQMNS LRAEDTALYYCAKDSRGYGDYSLGGAYWGQGTLVTVSSASTK GPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTS GVHTFP A VLQ S S GL YSL S SWT VP S S SLGTQT YICNVNHKP SNT KVDKKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMI SRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQ YNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISK AKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEW ESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFS CSVMHEALHNHYTQKSLSLSPGK (SEQ ID NO: 99) Sequence of the constant region of the heavy chain to CD3 (with Fc IgG4 WT) EVQLVESGGGLVQPGRSLRLSCAASGFTFHNYAMHWVRQAPG KGLEWVSGISWNSGSIGYADSVKGRFTISRDNAKNSLYLQMNS LRAEDTALYYCAKDSRGYGDYSLGGAYWGQGTLVTVSSASTK GPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTS GVHTFP AVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNT KVDKRVESKYGPPCPSCPAPEFLGGPSVFLFPPKPKDTLMISRTP EVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNST YRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQ PREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQ PENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMH EALHNHYTQKSLSLSLGK (SEQ ID NO: 100) Sequence of the heavy chain constant region to CD3 (with Fc-silenced IgG4) EVQLVESGGGLVQPGRSLRLSCAASGFTFHNYAMHWVRQAPG KGLEWVSGISWNSGSIGYADSVKGRFTISRDNAKNSLYLQMNS LRAEDTALYYCAKDSRGYGDYSLGGAYWGQGTLVTVSSASTK GPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTS GVHTFP AVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNT KVDKRVESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTP EVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNST YRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQ PREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQ PENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMH EALHNHYTQKSLSLSLGK (SEQ ID NO: 101)

In some embodiments, bispecific or multispecific antibodies are provided, which may have any of the configurations discussed herein, including, but not limited to, a bispecific three-chain antibody-like molecule. In some embodiments, a bispecific antibody may comprise at least one heavy chain variable region having binding specificity for CD22 and at least one heavy chain variable region having binding specificity for a protein other than CD22. In some embodiments, a bispecific antibody may comprise a heavy chain/light chain pair that has binding specificity for a first antigen, and a heavy chain from a heavy chain-only antibody comprising an Fc portion comprising CH2 and/or CH3 and/or CH4 domains, in the absence of a CH1 domain, and an antigen binding domain that binds an epitope of a second antigen or a different epitope of the first antigen. In one specific embodiment, the bispecific antibody comprises a heavy chain/light chain pair that has binding specificity for an antigen on an effector cell (e.g., the CD3 protein on a T

- 26 048216 cell), and a heavy chain from a heavy chain-only antibody containing an antigen-binding domain having binding specificity for CD22.

In some embodiments, when the binding compound of the invention is a bispecific antibody, one arm of the antibody (one binding fragment or one binding unit) is specific for human CD22, while the other arm may be specific for target cells, tumor-associated antigens, targeting antigens, such as integrins, etc., pathogen antigens, checkpoint proteins, etc. Target cells specifically include cancer cells, including, but not limited to, cells from hematological tumors, such as B-cell tumors, as described above. In some embodiments, one arm of the antibody (one binding fragment or one binding unit) is specific for human CD22, while the other arm is specific for CD3.

In some embodiments, the binding compound comprises an anti-CD3 light chain polypeptide comprising the sequence of SEQ ID NO: 92 linked to the sequence of SEQ ID NO: 97, an anti-CD3 heavy chain polypeptide comprising the sequence of any of SEQ ID NOs: 98, 99, 100, or 101, and an anti-CD22 heavy chain polypeptide comprising the sequence of any of SEQ ID NO: 2484 linked to the sequence of any of SEQ ID NOs: 93, 94, 95, or 96. These sequences can be combined in various ways to produce a bispecific antibody of a desired IgG subclass, e.g., IgG1, IgG4, silenced IgG1, silenced IgG4.

Various formats of bispecific antibodies are within the scope of the invention, including, but not limited to, single-chain polypeptides, double-chain polypeptides, triple-chain polypeptides, quadruple-chain polypeptides, and multiples thereof. Multispecific antibodies herein particularly include multispecific (e.g., bispecific) T-cell antibodies that bind to CD22 (anti-CD22 x CD3 antibodies), which are selectively expressed on mature B cells, and CD3. Such antibodies cause potent T-cell-mediated killing of cells expressing CD22.

Obtaining antibodies.

The multispecific binding compounds of the present invention can be prepared using methods known in the art. In a preferred embodiment, the heavy chain-only antibodies herein are produced by transgenic animals, including transgenic mice and rats, preferably rats in which the endogenous immunoglobulin genes are knocked out or disabled. In a preferred embodiment, the heavy chain-only antibodies of the present invention are produced in a UniRat™. The UniRat™ silences the endogenous immunoglobulin genes and utilizes the human immunoglobulin heavy chain translocus to express a diverse, naturally optimized repertoire of fully human HCAbs. While endogenous immunoglobulin loci in rats can be knocked out or silenced using a variety of technologies, zinc finger (endo)nuclease (ZNF) technology was used in UniRat™ to inactivate the endogenous rat J heavy chain locus, Ck light chain locus, and Ολ light chain locus. ZNF constructs for microinjection into oocytes can produce IgH and IgL knockout (KO) lines. For details, see, e.g., Geurts et al., 2009, Science 325:433. Characterization of Ig heavy chain knockout rats has been reported by Menoret et al., 2010, Eur. J. Immunol. 40:2932–2941. The advantages of ZNF technology are that non-homologous end joining for gene or locus silencing through deletions of up to a few kb can also provide a target site for homologous integration (Cui et al., 2011, Nat Biotechnol 29:64-67). The heavy chain-only human antibodies produced in UniRat™ are called UniAb™ and can bind epitopes that cannot be attacked by conventional antibodies. Their high specificity, affinity and small size make them ideal for mono- and polyspecific applications.

In addition to UniAb™, heavy chain-only antibodies lacking the camel backbone and VHH mutations and their functional VH regions are particularly included herein. Such heavy chain-only antibodies may, for example, be produced in transgenic rats or mice that contain gene loci consisting entirely of human heavy chains only, as described, for example, in WO2006/008548, but in other transgenic mammals such as rabbit, guinea pig, rats may also be used, with rats and mice being preferred. Heavy chain-only antibodies including their functional VHH or VH fragments may also be produced by recombinant DNA technology by expressing the encoding nucleic acid in a suitable eukaryotic or prokaryotic host, including, for example, mammalian cells (e.g., CHO cells), E. coli or yeast.

Heavy chain-only antibody domains combine the advantages of antibodies and small molecule drugs: they can be monovalent or multivalent; they have

- 27 048216 low toxicity; and cost-effective to manufacture. Due to their small size, these domains are easy to administer, including oral or topical administration, are characterized by high stability, including stability in the gastrointestinal tract; and their half-life can be tailored to the desired use or indication. In addition, the VH and VHH domains of HCAb can be produced in a cost-effective manner.

In a specific embodiment, the heavy chain-only antibodies of the invention, including UniAb™, have a native amino acid residue at the first position of the FR4 region (amino acid position 101 according to the Kabat numbering system) substituted with another amino acid residue that is capable of disrupting a surface-exposed hydrophobic patch comprising or associated with the native amino acid residue at that position. Such hydrophobic patches are typically buried at the interface with the constant region of the antibody light chain, but become surface-accessible in HCAb and serve, at least in part, to inhibit undesired aggregation and association of the HCAb light chain. The substituted amino acid residue is preferably charged, and more preferably positively charged, such as lysine (Lys, K), arginine (Arg, R) or histidine (His, H), preferably arginine (R). In a preferred embodiment, heavy chain-only antibodies derived from transgenic animals contain a Trp to Arg mutation at position 101. The resulting HCAbs preferably have high binding affinity for the antigen and are soluble under physiological conditions without aggregation.

In the present invention, human heavy chain-only anti-CD22 antibodies with unique sequences from UniRat™ animals (UniAb™) that bind to human CD22 in ELISA-based protein and cell binding assays were identified. The identified heavy chain variable region (VH) sequences (see Table 2) are positive for binding to human CD22 protein and/or for binding to CD22+ cells, and all are negative for binding to cells that do not express CD22.

Heavy chain-only antibodies that bind to non-overlapping epitopes on the CD22 protein, such as UniAbs™, can be identified using competitive binding assays such as enzyme-linked immunosorbent assays (ELISAs) or flow cytometric competitive binding assays. For example, competition between known antibodies that bind to a target antigen and the antibody of interest can be used. Using this approach, a panel of antibodies can be divided into those that compete with the reference antibody and those that do not. Non-competing antibodies are identified as binding to a distinct epitope that does not overlap with the epitope bound by the reference antibody. Often, one antibody is captured, the antigen is bound, and a second, labeled (e.g., biotinylated) antibody is tested in an ELISA-based assay for its ability to bind the captured antigen. This can also be done using surface plasmon resonance (SPR) platforms including the ProteOn XPR36 (BioRad, Inc), Biacore 2000 and Biacore T200 (GE Healthcare Life Sciences), and the MX96 SPR imaging system (Ibis technologies BV), as well as other biolayer interferometry platforms such as the Octet Red384 and Octet HTX (ForteBio, Pall Inc). For more information, see the examples in this paper.

Typically, an antibody competes with a reference antibody if it causes about a 15-100% reduction in binding of the reference antibody to a target antigen, as determined by standard methods, such as the competitive binding assays described above. In various embodiments, the relative inhibition is at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95% or higher.

Pharmaceutical compositions, uses and methods of treatment.

Another aspect of the present invention is to provide pharmaceutical compositions comprising one or more multispecific binding compounds of the present invention in admixture with a suitable pharmaceutically acceptable carrier. Pharmaceutically acceptable carriers used herein include, but are not limited to, adjuvants, solid carriers, water, buffers or other carriers used in the art for storing therapeutic components or combinations thereof.

In one embodiment, the pharmaceutical composition comprises a heavy chain-only antibody (e.g., UniAb™) that binds to CD22. In another embodiment, the pharmaceutical composition comprises a multispecific (including bispecific) heavy chain-only antibody (e.g., UniAb™) with binding specificity for two or more non-overlapping epitopes on the CD22 protein. In a preferred embodiment, the far

- 28 048216 the meceutical composition comprises a polyspecific (including bispecific) antibody containing only heavy chains (e.g. UniAb™) with binding specificity to CD22 and with binding specificity to a binding target on an effector cell (e.g. a binding target on a T cell, such as, for example, the CD3 protein on a T cell).

Pharmaceutical compositions of antibodies useful in accordance with the present invention are prepared for storage by mixing proteins having the desired degree of purity with optional pharmaceutically acceptable carriers, excipients, or stabilizers (see, for example, Remington's Pharmaceutical Sciences 16th edition, Osol, A. Ed. (1980)), for example, as lyophilized formulations or aqueous solutions. Acceptable carriers, excipients, or stabilizers are nontoxic to recipients at the dosages and concentrations employed and include buffers such as phosphate, citrate, and other organic acids; antioxidants including ascorbic acid and methionine; preservatives (such as octadecyldimethylbenzyl ammonium chloride; hexamethonium chloride; benzalkonium chloride, benzethonium chloride; phenol, butyl or benzyl alcohol; alkyl parabens such as methyl or propyl paraben; catechol; resorcinol; cyclohexanol; 3-pentanol; and m-cresol); low molecular weight (less than about 10 residues) polypeptides; proteins such as serum albumin, gelatin or immunoglobulins; hydrophilic polymers such as polyvinylpyrrolidone; amino acids such as glycine, glutamine, asparagine, histidine, arginine or lysine; monosaccharides, disaccharides and other carbohydrates including glucose, mannose or dextrins; chelating agents such as EDTA; sugars such as sucrose, mannitol, trehalose, or sorbitol; salt-forming counterions such as sodium; metal complexes (e.g., Zn-protein complexes); and/or nonionic surfactants such as TWEEN™, PLURONICS™, or polyethyleneglycol (PEG).

Pharmaceutical compositions for parenteral administration are preferably sterile and substantially isotonic and manufactured under good manufacturing practice (GMP) conditions. The pharmaceutical compositions may be provided in unit dosage form (e.g., unit dosage). The composition depends on the chosen route of administration. The antibodies described herein may be administered by intravenous injection or infusion, or subcutaneously. For injection administration, the antibodies described herein may be formulated in aqueous solutions, preferably in physiologically compatible buffers to reduce discomfort at the injection site. The solution may contain carriers, excipients, or stabilizers as described above. Alternatively, the antibodies may be lyophilized for mixing with a suitable vehicle, such as sterile, pyrogen-free water, prior to use.

Antibody formulations are described, for example, in U.S. Patent No. 9,034,324. Similar formulations may be used for heavy chain-only antibodies, including UniAb™, of the present invention. Antibody formulations for subcutaneous administration are described, for example, in US20160355591 and US20160166689.

Methods of application.

The heavy chain-only anti-CD22 antibodies, multispecific antibodies, and pharmaceutical compositions described herein can be used to treat diseases and conditions characterized by CD22 expression, including, but not limited to, the conditions and diseases described further herein.

CD22 is a 135 kDa type I transmembrane protein that is expressed at low levels on pre-B cells and immature B cells, maximally on mature B cells, and is completely down-regulated on plasma cells (e.g., Walker et al., Immunology, 2008 Mar; 123(3) 314–25). CD22 is highly expressed in follicular (primary and secondary B-cell zones), mantle, and marginal zone B cells and is reported to be present in 60–80% of patient specimens with B-cell malignancies (Alderson et al., Clin. Cancer Res 2009; 15(3) February 11, 2009). Due to its observed expression in a number of hematological malignancies, CD22 is a promising target for antibody-based therapeutics.

In one aspect, heavy chain-only anti-CD22 antibodies (e.g., UniAbs™) and pharmaceutical compositions of the present invention can be used to treat hematological malignancies characterized by CD22 expression, including, but not limited to, diffuse large B-cell lymphoma (DLBCL), non-Hodgkin's lymphoma, B-cell chronic lymphocytic leukemia (CLL), and B-cell acute lymphoblastic leukemia (ALL).

Diffuse large B-cell lymphoma (DLBCL or DLBCL) is the most common form of non-Hodgkin lymphoma in adults (Blood 1997 89(11):3909-18), with an estimated annual incidence of 7 to 8 cases per 100,000 people per year in the United States and the United Kingdom. It is characterized as an aggressive cancer that can arise in almost any part of the body. The causes of DLBCL are not well understood, and it can arise from normal B cells as well as malignant transformation of other types of lymphoma or leukemia cells. Treatment approaches are generally

- 29 048216 include chemotherapy and radiation therapy, and result in an overall five-year survival rate in adults of about 58%. Although some monoclonal antibodies have shown promise for the treatment of DLBCL, consistent clinical efficacy has not yet been definitively demonstrated. Therefore, there is a great need for new therapies, including immunotherapy, for DLBCL.

In another aspect, heavy chain-only anti-CD22 antibodies (e.g., UniAb™) and pharmaceutical compositions of the present invention can be used to treat autoimmune disorders characterized by disease-causing B cells that express CD22, including, but not limited to, systemic lupus erythematosus (SLE), rheumatoid arthritis (RA), and multiple sclerosis (MS).

Effective doses of the compositions of the present invention for treating a disease vary depending on many different factors, including the route of administration, the site of administration, the physiological state of the patient, whether the patient is a human or an animal, whether other drugs are administered, and whether the treatment is prophylactic or therapeutic. Typically, the patient is a human, but non-human mammals can also be treated, such as domestic animals such as dogs, cats, horses, etc., laboratory mammals such as rabbits, mice, rats, etc., and the like. Medicinal dosages can be adjusted to optimize safety and effectiveness.

Dosage levels can be readily determined by a skilled physician and can be changed if necessary, for example, if the subject's response to therapy needs to change. The amount of active substance that can be combined with carrier materials to produce a single dosage form varies depending on the host being treated and the particular route of administration. Unit dosage forms typically contain from about 1 to about 500 mg of active ingredient.

In some embodiments, the therapeutic dosage of the agent may be from about 0.0001 to 100 mg/kg and more typically from 0.01 to 5 mg/kg of the host body weight. For example, the dosage may be 1 mg/kg body weight or 10 mg/kg body weight or in the range of 1-10 mg/kg. An exemplary treatment regimen includes administration once every two weeks or once a month or once every 3-6 months. The therapeutic substances of the present invention are typically administered multiple times. The intervals between single doses may be weekly, monthly or yearly. The intervals may also be irregular, as indicated by measuring the level of the therapeutic substance in the patient's blood. Alternatively, the therapeutic substances of the present invention may be administered as a sustained release composition, in which case less frequent administration is required. The dosage and frequency vary depending on the half-life of the polypeptide in the patient.

Typically, the compositions are prepared as injectables, either as liquid solutions or suspensions; solid forms suitable for solution in, or suspension in, liquid vehicles prior to injection may also be prepared. The pharmaceutical compositions described herein are suitable for intravenous or subcutaneous administration, directly or after reconstitution from a solid (i.e., lyophilized) composition. The preparation may also be emulsified or encapsulated in liposomes or microparticles such as polylactide, polyglycolide, or a copolymer to enhance the adjuvant effect, as described above. Langer, Science 249: 1527, 1990 and Hanes, Advanced Drug Delivery Reviews 28: 97-119, 1997. The agents of the invention may be administered in the form of a depot injection or an implant preparation, which may be formulated to provide sustained or pulsatile release of the active substance. Pharmaceutical compositions are generally formulated to be sterile, substantially isotonic, and in full compliance with all Good Manufacturing Practice (GMP) standards of the U.S. Food and Drug Administration.

The toxicity of the antibodies and antibody structures described herein can be determined by standard pharmaceutical procedures in cell cultures or experimental animals, for example, by determining the LD50 (the dose lethal to 50% of the population) or LD100 (the dose lethal to 100% of the population). The dose ratio between the toxic and therapeutic effect is the therapeutic index. Data obtained from these cell culture assays and animal studies can be used to formulate a dose range that is non-toxic for use in humans. The dosage of the antibodies described herein is preferably within a range of circulating concentrations that includes the effective dose with little or no toxicity. The dosage may vary within this range depending on the dose used and the route of administration used. The exact composition, route of administration and dosage can be individually selected by the physician taking into account the patient's condition.

Compositions for administration typically comprise an antibody or other agent (e.g., another ablative agent) dissolved in a pharmaceutically acceptable carrier, preferably an aqueous carrier. Various aqueous carriers may be used, such as buffered saline.

- 30 048216 tvor and the like. These solutions are sterile and generally do not contain undesirable substances. These compositions can be sterilized by conventional, well-known sterilization methods. The compositions can contain pharmaceutically acceptable auxiliary substances necessary to approximate them to physiological conditions, such as pH-regulating agents and buffering agents, toxicity-regulating agents and the like, for example sodium acetate, sodium chloride, potassium chloride, calcium chloride, sodium lactate and the like. The concentration of active agent in these formulations may vary widely and will be selected primarily on the basis of fluid volumes, viscosity, body weight, and the like, in accordance with the particular route of administration selected and the needs of the patient (e.g., Remington's Pharmaceutical Science (15th ed., 1980) and Goodman & Gillman, The Pharmacological Basis of Therapeutics (Hardman et al., eds., 1996)).

Also included within the scope of the invention are kits containing active agents and their compositions, inventions and instructions for use. The kit may additionally contain at least one additional component, such as a chemotherapeutic drug, etc. Kits typically include a label indicating the intended use of the contents of the kit. As used herein, the term label includes any written or recorded materials supplied in or with the kit, or otherwise accompanying the kit.

Now that the invention has been fully described, it will be apparent to those skilled in the art that various changes and modifications can be made without departing from the spirit or scope of the invention.

Examples

Materials and methods.

CD22 protein binding.

Kinetic binding experiments to determine antigen-antibody affinity were performed on an Octet QK-384 system (ForteBio) using biolayer interferometry. Human IgG Fc Capture (AHC) biosensors (Forte Bio, Part No: 18-5064) were hydrated in assay buffer (1x PBS, 0.1% BSA, 0.02% Tween-20, pH 7.2) and pretreated in 100 mM glycine, pH 1.5. A baseline was established in assay buffer for 120 seconds. The AHC biosensors were then immobilized with UniAb™ at a concentration of 5 μg/mL for 120 seconds. Another baseline (120 seconds) was established in assay buffer. They were then loaded into a 7-point 1:2 dilution series of human CD22 protein in assay buffer starting at 250 nM. The last well of the analyte column contained assay buffer only to test for non-specific binding between the buffer and the loaded biosensors and was used as a control well. Association was monitored for 600 seconds, followed by dissociation for 900 seconds. Data analysis was performed using Octet Data Analysis v9.0 (ForteBio). Binding kinetics were analyzed using a standard 1:1 binding model.

CD22 cell binding.

Binding to CD22-positive cells was assessed by flow cytometry (Guava easyCyte 8HT, EMD Millipore) using the Daudi cell line (ATCC). Briefly, 100,000 target cells were stained with serial dilutions of purified UniAb™ for 30 min at 4°C. After incubation, cells were washed twice with flow cytometry buffer (1X PBS, 1% BSA, 0.1% _NaN3 ) and stained with goat F(ab') ₂ anti-human IgG conjugated to R-phycoerythrin (PE) (Southern Biotech, Cat# 2042-09) to detect cell-bound antibodies. After 20 min incubation at 4°C, cells were washed twice with flow cytometry buffer and then mean fluorescence intensity (MFI) was measured by flow cytometry. EC50 values were calculated using GraphPad Prism 7. Binding to cynomolgus monkey CD22-positive cells was determined using the same protocol with the following modifications: target cells were CHO cells stably transfected to express the cynomolgus monkey CD22 extracellular domain and each antibody was tested at a single concentration (~1.7 μg/mL) so EC50 values were not calculated.

Example 1: Genetically engineered rats expressing heavy chain-only antibodies.

The human and rat IgH locus was constructed and assembled from several parts. This involved modifying and combining the rat C region genes downstream of human _JH , and then adding the human VH6-D segment upstream. Two BACs with separate human _VH gene clusters [BAC6 and BAC3] were then co-injected with a BAC called Georg encoding an assembled and modified region comprising human VH6, all D, all _JH , and modified rat CY2a/1/2b ( _ACH1 ).

Transgenic rats carrying artificial heavy chain immunoglobulin loci in a disordered configuration were obtained. The IgG2a(AC.'n1).. IgG1(AC.'n1).. IgG2b(AC.'n1) genes lack the _CH 1 segment. The IgE, IgA constant region genes and the 3'-enhancer were included in the BAC Georg. RT-PCR and serum analysis (ELISA) of transgenic rats revealed productive rearrangement of the transgenic immunoglobulin loci and expression of heavy chain-only antibodies of various isotypes.

- 31 048216

Transgenic rats were crossed with rats with mutated endogenous heavy chain and light chain loci previously described in U.S. Patent Publication No. 2009/0098134 A1. Analysis of these animals demonstrated inactivation of rat heavy and light chain immunoglobulin expression and high levels of expression of heavy chain-only antibodies with variable regions encoded by the human V, D, and J genes. Immunization of the transgenic rats resulted in high titer serum responses of antigen-specific heavy chain-only antibodies. These transgenic rats expressing heavy chain-only antibodies with the human VDJ region were named UniRat™.

Example 2. Immunization.

Immunization with recombinant CD22 extracellular domain.

Twelve UniRat animals (6 HC27, 6 HC28) were immunized with recombinant human CD22 protein. Animals were immunized according to a standard protocol using Titermax/Alhydrogel adjuvant. Recombinant CD22 extracellular domain was purchased from R&D Systems and diluted in sterile saline and combined with the adjuvant. The immunogen was combined with Titermax and Alhydrogel adjuvants. The first immunization (priming) with immunogen in Titermax was performed on the left and right paws. Subsequent booster immunizations were performed in the presence of Alhydrogel and three days before boost selection were performed with immunogen in PBS. Serum was collected from rats at the last bled to determine serum titers.

Results for serum titers.

A summary of serum titers is shown in Fig. 17. In the graphs shown in Fig. 17, each line represents an individual animal. The legends to the graphs show the identification number of each individual animal. Binding activity for an 8-point serum dilution series was tested by ELISA against Fc-coated huCD22, His-tagged huCD22, His-tagged rhesus CD22, and a non-target His-tagged protein. A range of serum reactivity to both human and rhesus CD22 was observed among this group of animals. A serum response to the His-tagged protein was also observed.

Example 3. Binding to CD22-expressing cell lines.

Fig. 16 shows the target binding activity of the heavy chain-only anti-CD22 antibodies described herein. Column 1 shows the clone identification number of the heavy chain-only anti-CD22 antibody. Column 2 shows the protein binding affinity (KD), measured in molar concentration. Column 3 shows the protein binding dissociation constant (Koff), measured in seconds. Column 4 shows the binding to Daudi cells, measured as a fold of signal over background SIF signal. Column 5 shows the binding to CHO cells stably expressing cynomolgus monkey CD22, measured as a fold of signal over background SIF signal. Column 6 shows the binding to CHO cells that do not express the CD22 protein, measured as a fold of signal over background SIF signal.

Example 4. T cell-mediated cytotoxicity of CD22-positive cells using resting human pan-T cells.

Unstimulated human T cells were incubated with CD22-positive cells (Daudi) and different concentrations of bispecific antibodies. After 48 h, cells were analyzed by flow cytometry to measure cytotoxicity. Cell culture supernatants were used to measure IL-2 cytokine release. Positive control antibody refers to an antibody that contains the same arm to CD22 but a stronger affinity arm to CD3. The results are shown in Fig. 1A and Fig. 1B.

Unstimulated human T cells were incubated with CD22-positive cells (SUDHL10) and various concentrations of bispecific antibodies. After 72 hours, cells were analyzed by flow cytometry to measure cytotoxicity. Cell culture supernatants were used to measure IL-2 cytokine release. The positive control antibody refers to an antibody that contains the same arm to CD22 but a stronger affinity arm to CD3. The results are shown in Fig. 2A and Fig. 2B.

Unstimulated human T cells were incubated with CD22-positive DL-BCL (RI-1) cell line and different concentrations of bispecific antibodies with different effector:target cell (E:T) ratios of 10:1, 5:1 or 1:1. After 72 h, cells were analyzed by flow cytometry to measure cytotoxicity. Cell culture supernatants were used to measure IL-2 cytokine release. Positive control antibody refers to an antibody that contains the same arm to CD22 but a stronger affinity arm to CD3. The data demonstrate that the % cytotoxicity is dependent on the E:T ratio. The results are shown in Fig. 3A and Fig. 3B.

Example 5. T-cell-mediated cytotoxicity of CD22-positive cells using activated human pan-T cells.

Activated human T cells were incubated with CD22-positive cells (Daudi and RI-1) or a CD22-negative cell line (K562) and different concentrations of bispecific

- 32,048,216 antibodies. Cell lysis was measured using calcein fluorescence reading. The bispecific binder CD22xCD3_F2F specifically induced lysis of CD22+ cells but not CD22-K562 cells. The positive control antibody refers to an antibody that contains the same arm to CD22 but a stronger affinity arm to CD3. The negative control refers to an antibody with a non-tumor-specific arm and the same arm to CD3 as anti-CD3_F2F. The results are shown in Fig. 4.

Example 6. Binding of bispecific antibodies against CD22 and CD3 to cells.

CD22-positive Daudi, Raji, Ramos cells and CD22-negative K562 cells were incubated with the bispecific antibodies. Cell binding was measured by flow cytometry using anti-human IgG secondary antibody reagent. The data demonstrate that the bispecific antibodies bind to CD22+ cells but not to CD22- cells. The positive control antibody refers to an antibody that contains the same arm to CD22 but a stronger affinity arm to CD3. The negative control refers to an antibody with a non-tumor-specific arm and the same arm to CD3 as anti-CD3_F2F. The results are shown in Fig. 5.

Example 7. In vivo efficacy study with CD22-1 x CD3F2F in Daudi xenografts.

To test the in vivo efficacy of CD22-1 x CD3F2F, different doses of CD22-1 x CD3_F2F were administered to female NSG mice implanted with Daudi cells (5e6 cells/mouse) as shown in Fig. 6. The treatment schedule is shown below in Table 8. The mean tumor volume, body weight, percentage change in body weight, and individual tumor volume were used to evaluate the treatment efficacy.

Table 8 Approximate treatment regimen

Group Treatment Dose Method of administration η Scheme 1 PBS — v/b 10 2 CD22-1 x CD3 F2F 0.05 (1 µg/mouse) v/b 10 2 times a week (every 3-4 days) x 4 3 CD22-1 x CD3 F2F 0.5 (10 mcg/mouse) v/b 10 2 times a week (every 3-4 days) x 4 4 CD22-1 x CD3 F2F 2.5 (50 mcg/mouse) v/b 10 2 times a week (every 3-4 days) x 4 5 BiTe to CD19 x CD3 0.05 (1 µg/mouse) v/b 10 daily x10 6 Rituximab 15 (300 mcg/mouse) v/b 10 2 times a week (every 3-4 days) x 4

The CD22-1 x CD3F2F data are shown in Fig. 7-13, compared with the negative control and rituximab, and demonstrate the efficacy of CD22-1 x CD3_F2F.

Although preferred embodiments of the present invention have been shown and described herein, it will be apparent to those skilled in the art that these embodiments are provided for illustrative purposes only. Many variations, changes and substitutions will be apparent to those skilled in the art without departing from the scope and spirit of the present invention. It will be understood that various alternative embodiments of the invention described herein may be used in practicing the present invention. It is intended that the following claims define the scope of the invention and that methods and structures within the scope of these claims and their equivalents are thereby covered.

Sequence Listing <110> TENEOBIO, INC.

<120> POLYSPECIFIC ANTIBODIES CONTAINING ONLY HEAVY CHAINS THAT BIND TO CD22 AND CD3 <130> TNO-0017-WO <140> PCT/US2020/037566 <141> 06/12/2020 <150> 62/861708 <151> 06/14/2019 <160> 107 <170> PatentIn version 3.5

- 33 048216 <210> 1 <211> 10 <212> Protein <213> Artificial sequence <220>

<221> source <223> /note="Description of the artificial sequence: Synthetic peptide" <400> 1

Gly Asp Ser Ile Ser Ser Gly Asp Tyr Tyr

5 10 <210> 2 <211> 10 <212> Protein <213> Artificial sequence <220>

<221> source <223> /note="Description of artificial sequence: Synthetic peptide" <400> 2

Gly Asp Ser Ile Ser Ser Gly Gly Tyr Tyr

5 10 <210> 3 <211> 10 <212> Protein <213> Artificial sequence <220>

<221> source <223> /note="Description of the artificial sequence: Synthetic peptide" <400> 3

Gly Gly Ser Ile Ser Ser Gly Asp Tyr Tyr

5 10 <210> 4 <211> 10 <212> Protein <213> Artificial sequence <220>

<221> source <223> /note="Description of artificial sequence: Synthetic peptide" <400> 4

Gly Gly Ser Ile Ser Ser Ser Ser Tyr Tyr

5 10 <210> 5 <211> 8

- 34 048216 <212> Protein <213> Artificial sequence <220>

<221> source <223> /note="Description of the artificial sequence: Synthetic peptide" <400> 5

Gly Gly Ser Phe Ser Gly Tyr Tyr

5 <210> 6 <211> 10 <212> Protein <213> Artificial sequence <220>

<221> source <223> /note="Description of the artificial sequence: Synthetic peptide" <400> 6

Gly Asp Ser Ile Ser Ser Ser Ser Tyr Tyr

5 10 <210> 7 <211> 10 <212> Protein <213> Artificial sequence <220>

<221> source <223> /note="Description of the artificial sequence: Synthetic peptide" <400> 7

Gly Gly Ser Ile Thr Ser Ser Ser Tyr Tyr

5 10 <210> 8 <211> 10 <212> Protein <213> Artificial sequence <220>

<221> source <223> /note="Description of artificial sequence: Synthetic peptide" <400> 8

Gly Gly Ser Ile Ser Ser Ser Ser His Tyr

5 10 <210> 9 <211> 10 <212> Protein <213> Artificial sequence

- 35 048216 <220>

<221> source <223> /note="Description of the artificial sequence: Synthetic peptide" <400> 9

Gly Gly Ser Ile Ile Ser Ser Ser Tyr Tyr

5 10 <210> 10 <211> 10 <212> Protein <213> Artificial sequence <220>

<221> source <223> /note="Description of artificial sequence: Synthetic peptide" <400> 10

Gly Gly Ser Ile Asn Asp Asn Ser His Tyr

5 10 <210> 11 <211> 7 <212> Protein <213> Artificial sequence <220>

<221> source <223> /note="Description of the artificial sequence: Synthetic peptide" <400> 11

Ile Tyr Tyr Ser Gly Val Thr

5 <210> 12 <211> 7 <212> Protein <213> Artificial sequence <220>

<221> source <223> /note="Description of artificial sequence: Synthetic peptide" <400> 12

Ile Tyr Tyr Ser Gly Ala Thr

5 <210> 13 <211> 7 <212> Protein <213> Artificial sequence <220>

<221> source <223> /note="Description of the artificial sequence: Synthetic

- 36 048216 peptide" <400> 13

Ile Tyr Tyr Ser Gly Ser Thr

5 <210> 14 <211> 7 <212> Protein <213> Artificial sequence <220>

<221> source <223> /note="Description of artificial sequence: Synthetic peptide" <400> 14

Ile Tyr Tyr Thr Gly Ser Thr

5 <210> 15 <211> 7 <212> Protein <213> Artificial sequence <220>

<221> source <223> /note="Description of artificial sequence: Synthetic peptide" <400> 15

Val Tyr Tyr Thr Gly Ala Thr

5 <210> 16 <211> 7 <212> Protein <213> Artificial sequence <220>

<221> source <223> /note="Description of artificial sequence: Synthetic peptide" <400> 16

Ile His Tyr Ser Gly Ser Thr

5 <210> 17 <211> 7 <212> Protein <213> Artificial sequence <220>

<221> source <223> /note="Description of artificial sequence: Synthetic peptide" <400> 17

- 37 048216

Ile Tyr Tyr Ser Gly Ser Ala 1 5 <210> 18 <211> 10 <212> Protein <213> Artificial sequence <220>

<221> source <223> /note="Description of artificial sequence: Synthetic peptide" <400> 18 Thr Arg Glu Asp Ser Ser Asn Trp Arg Ser 1 5 10 <210> 19 <211> 10 <212> Protein <213> Artificial sequence <220>

<221> source <223> /note="Description of artificial sequence: Synthetic peptide" <400> 19 Thr Arg Asp Asp Ser Ser Asn Trp Arg Ser 1 5 10 <210> 20 <211> 10 <212> Protein <213> Artificial sequence <220>

<221> source <223> /note="Description of artificial sequence: Synthetic peptide" <400> 20 Thr Arg Glu Asp Ser Ser Ser Trp Arg Ser 1 5 10 <210> 21 <211> 10 <212> Protein <213> Artificial sequence <220>

<221> source <223> /note="Description of the synthetic sequence: Synthetic peptide" <400> 21 Ala Arg Glu Asp Ser Ser Ser Trp Arg Ser 1 5 10

- 38 048216 <210> 22 <211> 10 <212> Protein <213> Artificial sequence <220>

<221> source <223> /note="Description of artificial sequence: Synthetic peptide" <400> 22

Lys Arg Asp Asp Ser Ser Asn Trp Arg Ser

5 10 <210> 23 <211> 10 <212> Protein <213> Artificial sequence <220>

<221> source <223> /note="Description of the artificial sequence: Synthetic peptide" <400> 23

Ala Arg Asp Asp Ser Ser Asn Trp Arg Ser

5 10 <210> 24 <211> 118 <212> Protein <213> Artificial sequence <220>

<221> source <223> /note="Description of Artificial Sequence: Synthetic Polypeptide" <400> 24

Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu

5 1015

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Asp Ser Ile Ser Ser Gly 20 2530

Asp Tyr Tyr Trp Gly Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu 35 4045

Trp Ile Gly His Ile Tyr Tyr Ser Gly Val Thr Tyr Tyr Asn Pro Ser 50 5560

Leu Lys Ser Arg Val Thr Ile Ser Val Asp Thr Ser Arg Asn Gln Phe 65 70 7580

Ser Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr

- 39 048216

Cys Thr Arg Glu Asp Ser Ser Asn Trp Arg Ser Arg Gly Gln Gly Thr

100 105 110

Leu Val Thr Val Ser Ser

115 <210> 25 <211> 118 <212> Protein <213> Artificial sequence <220>

<221> source <223> /note="Description of Artificial Sequence: Synthetic Polypeptide" <400> 25

Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu

5 1015

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Asp Ser Ile Ser Ser Gly 20 2530

Asp Tyr Tyr Trp Gly Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu 35 4045

Trp Ile Gly His Ile Tyr Tyr Ser Gly Ala Thr Tyr Tyr Asn Pro Ser 50 5560

Leu Glu Asn Arg Val Thr Ile Ser Val Asp Thr Ser Lys Asn Gln Phe 65 70 7580

Ser Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr 85 9095

Cys Thr Arg Asp Asp Ser Ser Asn Trp Arg Ser Arg Gly Gln Gly Thr

100 105110

Leu Val Thr Val Ser Ser

115 <210> 26 <211> 118 <212> Protein <213> Artificial sequence <220>

<221> source <223> /note="Description of Artificial Sequence: Synthetic Polypeptide"

- 40 048216 <400> 26

Gln Leu Gln Leu 1

Gln Glu Ser Gly 5

Pro Gly Leu Val 10

Lys Pro Ser Glu 15

Th Leu Ser Leu

Thr Cys Thr Val

Ser Gly Asp Ser 25

Ile Ser Ser Gly 30

Asp Tyr Tyr Trp 35

Gly Trp Ile Arg 40

Gln Pro Pro Gly

Lys Gly Leu Glu 45

Trp Ile Gly His 50

Ile Tyr Tyr Ser 55

Gly Val Thr Tyr

Tyr Asn Pro Ser

Leu Lys Asn Arg 65

Val Thr Ile Ser

Val Asp Thr Ser 75

Lys Asn Gln Phe

Sir Leu Lys Leu

Ser Ser Val Thr 85

Ala Ala Asp Thr 90

Ala Val Tyr Tyr 95

CysThrArgGlu

100

Asp Ser Ser Ser

Trp Arg Ser Arg 105

Gly Gln Gly Thr 110

Leu Val Thr Val Ser Ser

115 <210> 27 <211> 118 <212> Protein <213> Artificial sequence <220>

<221> source <223> /note="Description of Artificial Sequence: Synthetic Polypeptide" <400> 27

Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu

5 1015

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Asp Ser Ile Ser SerGly

2530

Asp Tyr Tyr Trp Gly Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu 35 4045

Trp Ile Gly His Ile Tyr Tyr Ser Gly Val Thr Tyr Tyr Asn Pro Ser

5560

Leu Lys Asn Arg Val Thr Ile Ser Val Asp Thr Ser Arg Asn Gln Phe 65 70 7580

- 41 048216

Ser Leu Asn Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr 85 90 95

Cys Thr Arg Glu Asp Ser Ser Ser Trp Arg Ser Arg Gly Gln Gly Thr

100 105 110

Leu Val Thr Val Ser Ser

115 <210> 28 <211> 118 <212> Protein <213> Artificial sequence <220>

<221> source <223> /note="Description of Artificial Sequence: Synthetic Polypeptide" <400> 28

Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu

5 1015

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Asp Ser Ile Ser SerGly

2530

Asp Tyr Tyr Trp Gly Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu 35 4045

Trp Ile Gly His Ile Tyr Tyr Ser Gly Ala Thr Tyr Tyr Asn Pro Ser

5560

Leu Lys Asn Arg Val Thr Ile Ser Val Asp Thr Ser Arg Asn GlnPhe

70 7580

Ser Leu Asn Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val TyrTyr

9095

Cys Thr Arg Asp Asp Ser Ser Asn Trp Arg Ser Arg Gly Gln Gly Thr 100 105110

Leu Val Thr Val Ser Ser

115 <210> 29 <211> 118 <212> Protein <213> Artificial sequence <220>

- 42 048216 <221> source <223> /note="Description of artificial sequence: Synthetic polypeptide" <400> 29

Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu

5 1015

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Asp Ser Ile Ser SerGly

2530

Asp Tyr Tyr Trp Gly Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu 35 4045

Trp Ile Gly His Ile Tyr Tyr Ser Gly Ala Thr Tyr Tyr Asn Pro Ser 50 5560

Leu Lys Asn Arg Val Thr Ile Ser Val Asp Thr Ser Arg Asn Gln Phe 65 70 7580

Ser Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr 85 9095

Cys Thr Arg Asp Asp Ser Ser Asn Trp Arg Ser Arg Gly Gln Gly Thr

100 105110

Leu Val Thr Val Ser Ser

115 <210> 30 <211> 118 <212> Protein <213> Artificial sequence <220>

<221> source <223> /note="Description of Artificial Sequence: Synthetic Polypeptide" <400> 30

Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu

5 1015

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Asp Ser Ile Ser SerGly

2530

Asp Tyr Tyr Trp Gly Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu 35 4045

Trp Ile Gly His Ile Tyr Tyr Ser Gly Ala Thr Tyr Tyr Asn Pro Ser

5560

- 43 048216

Leu Lys Asn Arg Val Thr Ile Ser Val Asp Thr 65 70 75

Ser Lys Asn Gln Phe

Sir Leu Lys Leu

Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr

90 95

CysThrArgAspAsp

100

Ser Ser Asn Trp Arg Ser Arg Gly Gln Gly Thr

105 110

Leu Val Thr Val Ser Ser

115 <210> 31 <211> 118 <212> Protein <213> Artificial sequence <220>

<221> source <223> /note="Description of Artificial Sequence: Synthetic Polypeptide" <400> 31

Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu

5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Asp Ser Ile Ser Ser Gly 20 25 30

Gly Tyr Tyr Trp Gly Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu 35 40 45

Trp Ile Gly His Ile Tyr Tyr Ser Gly Ala Thr Tyr Tyr Asn Pro Ser 50 55 60

Leu Lys Asn Arg Val Thr Ile Ser Val Asp Thr Ser Lys Asn Gln Phe 65 70 75 80

Ser Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr 85 90 95

Cys Thr Arg Asp Asp Ser Ser Asn Trp Arg Ser Arg Gly Gln Gly Thr

100 105 110

Leu Val Thr Val Ser Ser

115 <210> 32 <211> 118 <212> Protein

- 44 048216 <213> Artificial sequence <220>

<221> source <223> /note="Description of Artificial Sequence: Synthetic Polypeptide" <400> 32

Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu

5 1015

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Asp Ser Ile Ser Ser Gly 20 2530

Asp Tyr Tyr Trp Gly Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu 35 4045

Trp Ile Gly His Ile Tyr Tyr Ser Gly Ala Thr Tyr Tyr Asn Pro Ser 50 5560

Leu Lys Asn Arg Val Thr Ile Ser Val Asp Thr Ser Arg Asn Gln Phe 65 70 7580

Ser Leu Asn Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr 85 9095

Cys Thr Arg Glu Asp Ser Ser Ser Trp Arg Ser Arg Gly Gln Gly Thr

100 105110

Leu Val Thr Val Ser Ser

115 <210> 33 <211> 118 <212> Protein <213> Artificial sequence <220>

<221> source <223> /note="Description of Artificial Sequence: Synthetic Polypeptide" <400> 33

Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu

5 1015

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Asp Ser Ile Ser SerGly

2530

Asp Tyr Tyr Trp Gly Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu

4045

- 45 048216

Trp Ile Gly His 50

Ile Tyr Tyr Ser 55

Gly Val Thr Tyr

Tyr Asn Pro Ser

Leu Lys Asn Arg 65

Val Thr Ile Ser

Val Asp Thr Ser

Lys Asn Gln Phe

Sir Leu Lys Leu

Ser Ser Val Thr 85

Ala Ala Asp Thr 90

Ala Val Tyr Tyr

CysThrArgAsp

100

Asp Ser Ser Asn

Trp Arg Ser Arg 105

Gly Gln Gly Thr 110

Leu Val Thr Val

115

Ser Ser <210> 34 <211> 118 <212> Protein <213> Artificial sequence <220>

<221> source <223> /note="Description of Artificial Sequence: Synthetic Polypeptide" <400> 34

Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu

5 1015

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Asp Ser Ile Ser Ser Gly 20 2530

Asp Tyr Tyr Trp Gly Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu 35 4045

Trp Ile Gly His Ile Tyr Tyr Ser Gly Val Thr Tyr Tyr Asn Pro Ser 50 5560

Leu Lys Asn Arg Val Thr Ile Ser Val Asp Thr Ser Arg Asn Gln Phe 65 70 7580

Ser Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr 85 9095

Cys Thr Arg Glu Asp Ser Ser Asn Trp Arg Ser Arg Gly Gln Gly Thr

100 105110

Leu Val Thr Val Ser Ser

115

- 46 048216 <210> 35 <211> 118 <212> Protein <213> Artificial sequence <220>

<221> source <223> /note="Description of Artificial Sequence: Synthetic Polypeptide" <400> 35

Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu

5 1015

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Asp Ser Ile Ser Ser Gly 20 2530

Asp Tyr Tyr Trp Gly Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu 35 4045

Trp Ile Gly His Ile Tyr Tyr Ser Gly Ala Thr Tyr Tyr Asn Pro Ser 50 5560

Leu Glu Asn Arg Val Thr Ile Ser Val Asp Thr Ser Lys Asn Gln Phe 65 70 7580

Ser Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr 85 9095

Cys Thr Arg Glu Asp Ser Ser Ser Trp Arg Ser Arg Gly Gln Gly Thr

100 105110

Leu Val Thr Val Ser Ser

115 <210> 36 <211> 118 <212> Protein <213>

Artificial sequence <220>

<221> source <223> /note="Description of Artificial Sequence: Synthetic Polypeptide" <400> 36

Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu

5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Asp Ser Ile Ser Ser Gly

25 30

Asp Tyr Tyr Trp Gly Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu

- 47 048216

Trp

Ile

Gly

His

Ile

Tyr

Tyr 55

Ser

Gly

Ala

Th

Tyr 60

Tyr

Asn

Pro

Ser

Leu

Lys

Asn

Arg

Val

Th

Ile

Ser

Val

Asp

Th 75

Ser

Arg

Asn

Gln

Phe 80

Ser

Leu

Lys

Leu

Ser 85

Ser

Val

Th

Ala

Ala 90

Asp

Th

Ala

Val

Tyr 95

Tyr

Cys

Th

Arg

Glu

100

Asp

Ser

Ser

Ser

Trp 105

Arg

Ser

Arg

Gly

Gln

110

Gly

Th

Leu Val Thr Val Ser Ser

115 <210> 37 <211> 118 <212> Protein <213> Artificial sequence <220>

<221> source <223> /note="Description of Artificial Sequence: Synthetic Polypeptide" <400> 37

Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu

5 1015

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Asp Ser Ile Ser Ser Gly 20 2530

Asp Tyr Tyr Trp Gly Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu 35 4045

Trp Ile Gly His Ile Tyr Tyr Ser Gly Val Thr Tyr Tyr Asn Pro Ser 50 5560

Leu Lys Asn Arg Val Thr Ile Ser Val Asp Thr Ser Lys Asn Gln Phe 65 70 7580

Ser Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr 85 9095

Cys Thr Arg Glu Asp Ser Ser Asn Trp Arg Ser Arg Gly Gln Gly Thr

100 105110

Leu Val Thr Val Ser Ser

- 48 048216

115 <210> 38 <211> 118 <212> Protein <213> Artificial sequence <220>

<221> source <223> /note="Description of Artificial Sequence: Synthetic Polypeptide" <400> 38

Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu

5 1015

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Asp Ser Ile Ser Ser Gly 20 2530

Asp Tyr Tyr Trp Gly Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu 35 4045

Trp Ile Gly His Ile Tyr Tyr Ser Gly Ala Thr Tyr Tyr Asn Pro Ser 50 5560

Leu Lys Asn Arg Val Thr Ile Ser Val Asp Thr Ser Arg Asn Gln Phe 65 70 7580

Ser Leu Asn Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr 85 9095

Cys Thr Arg Glu Asp Ser Ser Asn Trp Arg Ser Arg Gly Gln Gly Thr

100 105110

Leu Val Thr Val Ser Ser

115 <210> 39 <211> 118 <212> Protein <213> Artificial sequence <220>

<221> source <223> /note="Description of Artificial Sequence: Synthetic Polypeptide" <400> 39

Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu

5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Asp Ser Ile Ser Ser Gly

25 30

- 49 048216

Asp Tyr Tyr Trp Gly Trp Ile Arg

40

Gln Pro Pro Gly Lys Gly Leu Glu 45

Trp Ile Gly His Ile Tyr Tyr Ser

55

Gly Ala Thr Tyr Tyr Asn Pro Ser 60

Leu Lys Asn Arg Val Thr Ile Ser 65 70

Val Asp Thr Ser Lys Asn Gln Phe

80

Ser Leu Lys Leu Ser Ser Val Thr 85

Ala Ala Asp Thr Ala Val Tyr Tyr

95

Cys Thr Arg Glu Asp Ser Ser Asn

100

Trp Arg Ser Arg Gly Gln Gly Thr

105 110

Leu Val Thr Val Ser Ser

115 <210> 40 <211> 118 <212> Protein <213>

Artificial sequence <220>

<221> source <223> /note="Description of Artificial Sequence: Synthetic Polypeptide" <400> 40

Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu

5 1015

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Asp Ser Ile Ser Ser Gly 20 2530

Asp Tyr Tyr Trp Gly Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu 35 4045

Trp Ile Gly His Ile Tyr Tyr Ser Gly Ala Thr Tyr Tyr Asn Pro Ser 50 5560

Leu Lys Asn Arg Val Thr Ile Ser Val Asp Thr Ser Lys Asn Gln Phe 65 70 7580

Ser Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr 85 9095

Cys Thr Arg Glu Asp Ser Ser Ser Trp Arg Ser Arg Gly Gln Gly Thr

100 105110

- 50 048216

Leu Val Thr Val Ser Ser

115 <210> 41 <211> 118 <212> Protein <213> Artificial sequence <220>

<221> source <223> /note="Description of Artificial Sequence: Synthetic Polypeptide" <400> 41

Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu

5 1015

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Asp Ser Ile Ser SerGly

2530

Asp Tyr Tyr Trp Gly Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu 35 4045

Trp Ile Gly His Ile Tyr Tyr Ser Gly Ala Thr Tyr Tyr Asn Pro Ser 50 5560

Leu Lys Asn Arg Val Thr Ile Ser Val Asp Thr Ser Arg Asn GlnPhe

70 7580

Ser Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val TyrTyr

9095

Cys Thr Arg Glu Asp Ser Ser Asn Trp Arg Ser Arg Gly Gln Gly Thr

100 105110

Leu Val Thr Val Ser Ser

115 <210> 42 <211> 118 <212> Protein <213> Artificial sequence <220>

<221> source <223> /note="Description of Artificial Sequence: Synthetic Polypeptide" <400> 42

Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu

5 10 15

- 51 048216

Thr Leu Ser Leu Thr Cys Thr Val

Ser Gly Asp Ser Ile Ser Ser Gly 25 30

Asp Tyr Tyr Trp Gly Trp Ile Arg

40

Gln Pro Pro Gly Lys Gly Leu Glu 45

Trp Ile Gly Ser Ile Tyr Tyr Ser

55

Gly Ala Thr Tyr Tyr Asn Pro Ser 60

Leu Lys Asn Arg Val Thr Ile Ser 65 70

Val Asp Thr Ser Lys Asn Gln Phe

80

Ser Leu Lys Leu Ser Ser Val Thr 85

Ala Ala Asp Thr Ala Val Tyr Tyr

95

Cys Thr Arg Asp Asp Ser Ser Asn

100

Trp Arg Ser Arg Gly Gln Gly Thr

105 110

Leu Val Thr Val Ser Ser

115 <210> 43 <211> 118 <212> Protein <213> Artificial sequence <220>

<221> source <223> /note="Description of Artificial Sequence: Synthetic Polypeptide" <400> 43

Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu

5 1015

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Gly Ser Ile Ser SerGly

2530

Asp Tyr Tyr Trp Gly Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu 35 4045

Trp Ile Gly His Ile Tyr Tyr Ser Gly Ala Thr Tyr Tyr Asn Pro Ser

5560

Leu Lys Asn Arg Val Thr Ile Ser Val Asp Thr Ser Arg Asn GlnPhe

70 7580

Ser Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val TyrTyr

9095

- 52 048216

Cys Thr Arg Asp Asp Ser Ser Asn Trp Arg Ser Arg Gly Gln Gly Thr

100 105 110

Leu Val Thr Val Ser Ser

115 <210> 44 <211> 118 <212> Protein <213> Artificial sequence <220>

<221> source <223> /note="Description of Artificial Sequence: Synthetic Polypeptide" <400> 44

Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu

5 1015

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Gly Ser Ile Ser SerGly

2530

Asp Tyr Tyr Trp Gly Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu 35 4045

Trp Ile Gly His Ile Tyr Tyr Ser Gly Ala Thr Tyr Tyr Asn Pro Ser 50 5560

Leu Glu Asn Arg Val Thr Ile Ser Val Asp Thr Ser Lys Asn GlnPhe

70 7580

Ser Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val TyrTyr

9095

Cys Thr Arg Asp Asp Ser Ser Asn Trp Arg Ser Arg Gly Gln Gly Thr

100 105110

Leu Val Thr Val Ser Ser

115 <210> 45 <211> 118 <212> Protein <213> Artificial sequence <220>

<221> source <223> /note="Description of Artificial Sequence: Synthetic Polypeptide" <400> 45

- 53 048216

Gln Leu Gln Leu 1

Gln Glu Ser Gly 5

Pro Gly Leu Val 10

Lys Pro Ser Glu 15

Th Leu Ser Leu

Thr Cys Thr Val

Ser Gly Asp Ser 25

Ile Ser Ser Gly 30

Asp Tyr Tyr Trp 35

Gly Trp Ile Arg 40

Gln Pro Pro Gly

Lys Gly Leu Glu 45

Trp Ile Gly Ser 50

Ile Tyr Tyr Ser 55

Gly Ser Thr Tyr

Tyr Asn Pro Ser

Leu Lys Ser Arg 65

Val Thr Ile Ser

Val Asp Thr Ser 75

Lys Asn Gln Phe

Sir Leu Lys Leu

Ser Ser Val Thr 85

Ala Ala Asp Thr 90

Ala Val Tyr Tyr 95

CysThrArgGlu

100

Asp Ser Ser Ser

Trp Arg Ser Arg 105

Gly Gln Gly Th

110

Leu Val Thr Val Ser Ser

115 <210> 46 <211> 118 <212> Protein <213> Artificial sequence <220>

<221> source <223> /note="Description of Artificial Sequence: Synthetic Polypeptide" <400> 46

Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu

5 10 15

Th Leu Ser Leu

Thr Cys Thr Val

Ser Gly Asp Ser 25

Ile Ser Ser Gly 30

Asp Tyr Tyr Trp 35

Gly Trp Ile Arg 40

Gln Pro Pro Gly

Lys Gly Leu Glu 45

Trp Ile Gly Asn 50

Ile Tyr Tyr Ser 55

Gly Ala Thr Tyr

Tyr Asn Pro Ser

Leu Lys Asn Arg 65

Val Thr Ile Ser

Val Asp Thr Ser 75

Lys Asn Gln Phe

- 54 048216

Sir Leu Lys Leu

Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr

90 95

Cys

Thr Arg Glu Asp Ser Ser Ser Trp Arg Ser Arg Gly Gln Gly Thr

100 105 110

Leu Val Thr Val Ser Ser

115 <210> 47 <211> 118 <212> Protein <213> Artificial sequence <220>

<221> source <223> /note="Description of Artificial Sequence: Synthetic Polypeptide" <400> 47

Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu

5 1015

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Asp Ser Ile Ser SerGly

2530

Gly Tyr Tyr Trp Gly Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu 35 4045

Trp Ile Gly Ser Ile Tyr Tyr Ser Gly Ser Thr Tyr Tyr Asn Pro Ser 50 5560

Leu Lys Asn Arg Val Thr Ile Ser Val Asp Thr Ser Lys Asn GlnPhe

70 7580

Ser Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val TyrTyr

9095

Cys Thr Arg Glu Asp Ser Ser Ser Trp Arg Ser Arg Gly Gln Gly Thr

100 105110

Leu Val Thr Val Ser Ser

115 <210> 48 <211> 118 <212> Protein <213> Artificial sequence <220>

<221> source <223> /note="Description of the artificial sequence: Synthetic

- 55 048216 polypeptide" <400> 48

Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu

5 1015

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Asp Ser Ile Ser Ser Gly 20 2530

Asp Tyr Tyr Trp Gly Trp Ile Arg His Pro Pro Gly Lys Gly Leu Glu 35 4045

Trp Ile Gly His Ile Tyr Tyr Ser Gly Ala Thr Tyr Tyr Asn Pro Ser 50 5560

Leu Lys Asn Arg Val Thr Ile Ser Val Asp Thr Ser Lys Asn Gln Phe 65 70 7580

Ser Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr 85 9095

Cys Thr Arg Glu Asp Ser Ser Ser Trp Arg Ser Arg Gly Gln Gly Thr

100 105110

Leu Val Thr Val Ser Ser

115 <210> 49 <211> 118 <212> Protein <213> Artificial sequence <220>

<221> source <223> /note="Description of Artificial Sequence: Synthetic Polypeptide" <400> 49

Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu

5 1015

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Asp Ser Ile Ser SerGly

2530

Asp Tyr Tyr Trp Gly Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu 35 4045

Trp Ile Gly Asn Ile Tyr Tyr Ser Gly Ala Thr Tyr Tyr Asn Pro Ser

5560

Leu Lys Asn Arg Val Thr Ile Ser Val Asp Thr Ser Lys Asn Gln Phe

- 56 048216

Sir Leu Lys Leu

Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr

90 95

Cys

Thr Arg Glu Asp Ser Ser Asn Trp Arg Ser Arg Gly Gln Gly Thr

100 105 110

Leu Val Thr Val Ser Ser

115 <210> 50 <211> 118 <212> Protein <213> Artificial sequence <220>

<221> source <223> /note="Description of Artificial Sequence: Synthetic Polypeptide" <400> 50

Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu

5 1015

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Asp Ser Ile Ser Ser Gly 20 2530

Asp Tyr Tyr Trp Gly Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu 35 4045

Trp Ile Gly Asn Ile Tyr Tyr Ser Gly Ala Thr Tyr Tyr Asn Pro Ser 50 5560

Leu Lys Asn Arg Val Thr Ile Ser Val Asp Thr Ser Lys Asn Gln Phe 65 70 7580

Ser Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr 85 9095

Cys Thr Arg Asp Asp Ser Ser Asn Trp Arg Ser Arg Gly Gln Gly Thr

100 105110

Leu Val Thr Val Ser Ser

115 <210> 51 <211> 118 <212> Protein <213> Artificial sequence

- 57 048216 <220>

<221> source <223> /note="Description of Artificial Sequence: Synthetic Polypeptide" <400> 51

Gln Leu Gln Leu 1

Gln Glu Ser Gly 5

Pro Gly Leu Val 10

Lys Pro Ser Glu 15

Th Leu Ser Leu

Thr Cys Thr Val

Ser Gly Gly Ser 25

Ile Ser Ser Ser 30

Ser Tyr Tyr Trp 35

Gly Trp Ile Arg 40

Gln Pro Pro Gly

Lys Gly Leu Glu 45

Trp Ile Gly Ser 50

Ile Tyr Tyr Ser 55

Gly Ser Thr Tyr

Tyr Asn Pro Ser

Leu Lys Ser Arg 65

Val Thr Ile Ser

Val Asp Thr Ser 75

Lys Asn Gln Phe

Sir Leu Lys Leu

Ser Ser Val Thr 85

Ala Ala Asp Thr 90

Ala Val Tyr Tyr 95

CysThrArgAsp

100

Asp Ser Ser Asn

Trp Arg Ser Arg 105

Gly Gln Gly Thr 110

Leu Val Thr Val Ser Ser

115 <210> 52 <211> 118 <212> Protein <213> Artificial sequence <220>

<221> source <223> /note="Description of Artificial Sequence: Synthetic Polypeptide" <400> 52

Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu

5 1015

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Asp Ser Ile Ser SerGly

2530

Asp Tyr Tyr Trp Gly Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu 35 4045

Trp Ile Gly Ser Ile Tyr Tyr Ser Gly Ser Thr Tyr Tyr Asn Pro Ser

5560

- 58 048216

Leu Lys Ser Arg Val Thr Ile Ser Val Asp Thr 65 70 75

Ser Lys Asn Gln Phe

Ser

Leu

Lys

Leu

Ser 85

Ser

Val

Th

Ala

Ala 90

Asp

Th

Ala

Val

Tyr 95

Tyr

Cys

Th

Arg

Glu

100

Asp

Ser

Ser

Asn

Trp 105

Arg

Ser

Arg

Gly

Gln

110

Gly

Th

Leu Val Thr Val Ser Ser

115 <210> 53 <211> 118 <212> Protein <213> Artificial sequence <220>

<221> source <223> /note="Description of Artificial Sequence: Synthetic Polypeptide" <400> 53

Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu

5 1015

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Asp Ser Ile Ser Ser Gly 20 2530

Asp Tyr Tyr Trp Gly Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu 35 4045

Trp Ile Gly Ser Ile Tyr Tyr Thr Gly Ser Thr Tyr Tyr Asn Pro Ser 50 5560

Leu Lys Ser Arg Val Thr Ile Ser Val Asp Thr Ser Lys Asn Gln Phe 65 70 7580

Ser Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr 85 9095

Cys Thr Arg Glu Asp Ser Ser Asn Trp Arg Ser Arg Gly Gln Gly Thr

100 105110

Leu Val Thr Val Ser Ser

115 <210> 54 <211> 118

- 59 048216 <212> Protein <213> Artificial sequence <220>

<221> source <223> /note="Description of Artificial Sequence: Synthetic Polypeptide" <400> 54

Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu

5 1015

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Gly Ser Ile Ser SerGly

2530

Asp Tyr Tyr Trp Gly Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu 35 4045

Trp Ile Gly His Ile Tyr Tyr Ser Gly Ala Thr Tyr Tyr Asn Pro Ser

5560

Leu Lys Asn Arg Val Thr Ile Ser Val Asp Thr Ser Arg Asn GlnPhe

70 7580

Ser Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val TyrTyr

9095

Cys Thr Arg Glu Asp Ser Ser Asn Trp Arg Ser Arg Gly Gln Gly Thr 100 105110

Leu Val Thr Val Ser Ser

115 <210> 55 <211> 116 <212> Protein <213> Artificial sequence <220>

<221> source <223> /note="Description of Artificial Sequence: Synthetic Polypeptide" <400> 55

Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu

5 1015

Thr Leu Ser Leu Thr Cys Ala Val Tyr Gly Gly Ser Phe Ser GlyTyr

2530

Tyr Trp Gly Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp Ile

4045

- 60 048216

Gly His Ile Tyr Tyr Ser Gly Ala

55

Thr Tyr Tyr Asn Pro Ser Leu Lys 60

Asn Arg Val Thr Ile Ser Val Asp Thr Ser Arg Asn Gln Phe Ser Leu 65 70 75 80

Asn Leu

Ser

Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys Thr

90 95

Arg Glu Asp Ser

100

Ser Ser Trp Arg Ser Arg Gly Gln Gly Thr Leu Val

105 110

Thr Val Ser Ser

115 <210> 56 <211> 118 <212> Protein <213> Artificial sequence <220>

<221> source <223> /note="Description of Artificial Sequence: Synthetic Polypeptide" <400> 56

Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu

5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Gly Ser Ile Ser Ser Ser 20 25 30

Ser Tyr Tyr Trp Gly Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu 35 40 45

Trp Ile Gly His Ile Tyr Tyr Ser Gly Val Thr Tyr Tyr Asn Pro Ser 50 55 60

Leu Lys Asn Arg Val Thr Ile Ser Val Asp Thr Ser Lys Asn Gln Phe 65 70 75 80

Ser Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Ala Tyr Tyr 85 90 95

Cys Thr Arg Asp Asp Ser Ser Asn Trp Arg Ser Arg Gly Gln Gly Thr

100 105 110

Leu Val Thr Val Ser Ser

115

- 61 048216 <210> 57 <211> 118 <212> Protein <213> Artificial sequence <220>

<221> source <223> /note="Description of Artificial Sequence: Synthetic Polypeptide" <400> 57

Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu

5 1015

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Gly Ser Ile Ser Ser Ser 20 2530

Ser Tyr Tyr Trp Gly Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu 35 4045

Trp Ile Gly His Ile Tyr Tyr Ser Gly Val Thr Tyr Tyr Asn Pro Ser 50 5560

Leu Lys Asn Arg Val Thr Ile Ser Val Asp Thr Ser Lys Asn Gln Phe 65 70 7580

Ser Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr 85 9095

Cys Thr Arg Asp Asp Ser Ser Asn Trp Arg Ser Arg Gly Gln Gly Thr

100 105110

Leu Val Thr Val Ser Ser

115 <210> 58 <211> 118 <212> Protein <213> Artificial sequence <220>

<221> source <223> /note="Description of Artificial Sequence: Synthetic Polypeptide" <400> 58

Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu

5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser

25

Gly Gly Ser Ile Ser Ser Ser

- 62 048216

Ser Tyr Tyr Trp Gly Trp Ile Arg

40

Gln Pro Pro Gly Lys Gly Leu Glu 45

Trp Ile Gly Ser Ile Tyr Tyr Ser

55

Gly Ser Thr Tyr Tyr Asn Pro Ser 60

Leu Lys Ser Arg Val Thr Ile Ser 65 70

Val Asp Thr Ser Arg Asn Gln Phe

80

Ser Leu Asn Leu Ser Ser Val Thr

Ala Ala Asp Thr Ala Val Tyr Tyr

95

Cys Thr Arg Glu Asp Ser Ser Ser

100

Trp Arg Ser Arg Gly Gln Gly Thr

105 110

Leu Val Thr Val Ser Ser

115 <210> 59 <211> 118 <212> Protein <213> Artificial sequence <220>

<221> source <223> /note="Description of Artificial Sequence: Synthetic Polypeptide" <400> 59

Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu

5 1015

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Asp Ser Ile Ser SerSer

2530

Ser Tyr Tyr Trp Gly Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu 35 4045

Trp Ile Gly Ser Ile Tyr Tyr Ser Gly Ser Thr Tyr Tyr Asn Pro Ser 50 5560

Leu Lys Ser Arg Val Thr Ile Ser Val Asp Thr Ser Lys Asn Gln Phe 65 70 7580

Ser Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr 85 9095

Cys Thr Arg Glu Asp Ser Ser Asn Trp Arg Ser Arg Gly Gln Gly Thr

100 105110

- 63 048216

Leu Val Thr Val Ser Ser

115 <210> 60 <211> 118 <212> Protein <213> Artificial sequence <220>

<221> source <223> /note="Description of Artificial Sequence: Synthetic Polypeptide" <400> 60

Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu

5 1015

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Gly Ser Ile Ser SerSer

2530

Ser Tyr Tyr Trp Gly Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu 35 4045

Trp Ile Gly Ser Ile Tyr Tyr Ser Gly Ser Thr Tyr Tyr Asn Pro Ser

5560

Leu Lys Ser Arg Val Thr Ile Ser Val Asp Thr Ser Lys Asn GlnPhe

70 7580

Ser Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val TyrTyr

9095

Cys Ala Arg Glu Asp Ser Ser Ser Trp Arg Ser Arg Gly Gln Gly Thr 100 105110

Leu Val Thr Val Ser Ser

115 <210> 61 <211> 118 <212> Protein <213> Artificial sequence <220>

<221> source <223> /note="Description of Artificial Sequence: Synthetic Polypeptide" <400> 61

Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu

5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Gly Ser Ile Ser Ser Ser

- 64 048216

Ser

Tyr

Tyr 35

Trp

Gly

Trp

Ile

Arg 40

Gln

Pro

Pro

Gly

Lys 45

Gly

Leu

Glu

Trp

Number 50

Gly

Ser

Ile

Tyr

Tyr 55

Ser

Gly

Ala

Th

Tyr 60

Tyr

Asn

Pro

Ser

Leu 65

Lys

Asn

Arg

Val

Th 70

Ile

Ser

Val

Asp

Th 75

Ser

Lys

Asn

Gln

Phe 80

Ser

Leu

Lys

Leu

Ser 85

Ser

Val

Th

Ala

Ala 90

Asp

Th

Ala

Val

Tyr 95

Tyr

Cys

Th

Arg

Glu

100

Asp

Ser

Ser

Ser

Trp 105

Arg

Ser

Arg

Gly

Gln

110

Gly

Th

Leu

Val

Th

115

Val

Ser

Ser <210> 62 <211> 118 <212> Protein <213>

Artificial sequence <220>

<221> source <223> /note="Description of Artificial Sequence: Synthetic Polypeptide" <400> 62

Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu

5 1015

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Gly Ser Ile Ser Ser Ser 20 2530

Ser Tyr Tyr Trp Gly Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu 35 4045

Trp Ile Gly His Ile Tyr Tyr Ser Gly Ala Thr Tyr Tyr Asn Pro Ser 50 5560

Leu Lys Asn Arg Val Thr Ile Ser Val Asp Thr Ser Arg Asn Gln Phe 65 70 7580

Ser Leu Asn Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr 85 9095

Cys Thr Arg Glu Asp Ser Ser Ser Trp Arg Ser Arg Gly Gln Gly Thr

- 65 048216

100

105

110

Leu Val Thr Val Ser Ser

115 <210> 63 <211> 118 <212> Protein <213> Artificial sequence <220>

<221> source <223> /note="Description of Artificial Sequence: Synthetic Polypeptide" <400> 63

Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu

5 1015

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Gly Ser Ile Ser Ser Ser 20 2530

Ser Tyr Tyr Trp Gly Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu 35 4045

Trp Ile Gly Ser Ile Tyr Tyr Ser Gly Ser Thr Tyr Tyr Asn Pro Ser 50 5560

Leu Lys Ser Arg Val Thr Ile Ser Val Asp Thr Ser Lys Asn Gln Phe 65 70 7580

Ser Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr 85 9095

Cys Thr Arg Glu Asp Ser Ser Ser Trp Arg Ser Arg Gly Gln Gly Thr

100 105110

Leu Val Thr Val Ser Ser

115 <210> 64 <211> 118 <212> Protein <213> Artificial sequence <220>

<221> source <223> /note="Description of Artificial Sequence: Synthetic Polypeptide" <400> 64

Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu

5 10 15

- 66 048216

Thr Leu Ser Leu Thr Cys Thr Val

Ser Gly Gly Ser Ile Ser Ser Ser 25 30

Ser Tyr Tyr Trp Gly Trp Ile Arg

40

Gln Pro Pro Gly Lys Gly Leu Glu 45

Trp Ile Gly His Ile Tyr Tyr Ser

55

Gly Ala Thr Tyr Tyr Asn Pro Ser 60

Leu Lys Asn Arg Val Thr Ile Ser 65 70

Val Asp Thr Ser Lys Asn Gln Phe

80

Ser Leu Lys Leu Ser Ser Val Thr 85

Ala Ala Asp Thr Ala Val Tyr Tyr

95

Cys Thr Arg Asp Asp Ser Ser Asn

100

Trp Arg Ser Arg Gly Gln Gly Thr

105 110

Leu Val Thr Val Ser Ser

115 <210> 65 <211> 118 <212> Protein <213> Artificial sequence <220>

<221> source <223> /note="Description of Artificial Sequence: Synthetic Polypeptide" <400> 65

Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu

5 1015

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Gly Ser Ile Ser Ser Ser 20 2530

Ser Tyr Tyr Trp Gly Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu 35 4045

Trp Ile Gly Ser Ile Tyr Tyr Ser Gly Ala Thr Tyr Tyr Asn Pro Ser 50 5560

Leu Lys Asn Arg Val Thr Ile Ser Val Asp Thr Ser Arg Asn Gln Phe 65 70 7580

Ser Leu Asn Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr 85 9095

- 67 048216

Cys Thr Arg Glu Asp Ser Ser Ser Trp Arg Ser Arg Gly Gln Gly Thr

100 105 110

Leu Val Thr Val Ser Ser

115 <210> 66 <211> 118 <212> Protein <213> Artificial sequence <220>

<221> source <223> /note="Description of Artificial Sequence: Synthetic Polypeptide" <400> 66

Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu

5 1015

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Gly Ser Ile Ser Ser Ser 20 2530

Ser Tyr Tyr Trp Gly Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu 35 4045

Trp Ile Gly Ser Ile Tyr Tyr Ser Gly Ser Thr Tyr Tyr Asn Pro Ser 50 5560

Leu Lys Asn Arg Val Thr Ile Ser Val Asp Thr Ser Lys Asn Gln Phe 65 70 7580

Ser Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr 85 9095

Cys Thr Arg Glu Asp Ser Ser Asn Trp Arg Ser Arg Gly Gln Gly Thr

100 105110

Leu Val Thr Val Ser Ser

115 <210> 67 <211> 118 <212> Protein <213> Artificial sequence <220>

<221> source <223> /note="Description of Artificial Sequence: Synthetic Polypeptide"

- 68 048216 <400> 67

Gln Leu Gln Leu 1

Gln Glu Ser Gly 5

Pro Gly Leu Val 10

Lys Pro Ser Glu 15

Th Leu Ser Leu

Thr Cys Thr Val

Ser Gly Gly Ser 25

Ile Thr Ser Ser 30

Ser Tyr Tyr Trp 35

Gly Trp Ile Arg 40

Gln Pro Pro Gly

Lys Gly Leu Glu 45

Trp Ile Gly Ser 50

Ile Tyr Tyr Ser 55

Gly Ser Thr Tyr

Tyr Asn Pro Ser

Leu Lys Ser Arg 65

Val Thr Ile Ser

Val Asp Thr Ser 75

Lys Asn Gln Phe

Sir Leu Lys Leu

Ser Ser Val Thr 85

Ala Ala Asp Thr 90

Ala Val Tyr Tyr 95

CysThrArgAsp

100

Asp Ser Ser Asn

Trp Arg Ser Arg 105

Gly Gln Gly Thr 110

Leu Val Thr Val Ser Ser

115 <210> 68 <211> 118 <212> Protein <213> Artificial sequence <220>

<221> source <223> /note="Description of Artificial Sequence: Synthetic Polypeptide" <400> 68

Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu

5 1015

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Gly Ser Ile Ser Ser Ser 20 2530

Ser Tyr Tyr Trp Gly Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu 35 4045

Trp Ile Gly Ser Ile Tyr Tyr Ser Gly Val Thr Tyr Tyr Asn Pro Ser 50 5560

Leu Lys Asn Arg Val Thr Ile Ser Val Asp Thr Ser Lys Asn Gln Phe 65 70 7580

- 69 048216

Ser Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr 85 90 95

Cys Thr Arg Glu Asp Ser Ser Asn Trp Arg Ser Arg Gly Gln Gly Thr

100 105 110

Leu Val Thr Val Ser Ser

115 <210> 69 <211> 118 <212> Protein <213> Artificial sequence <220>

<221> source <223> /note="Description of Artificial Sequence: Synthetic Polypeptide" <400> 69

Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu

5 1015

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Gly Ser Ile Ser Ser Ser 20 2530

Ser Tyr Tyr Trp Gly Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu 35 4045

Trp Ile Gly Ser Ile Tyr Tyr Ser Gly Ser Thr Tyr Tyr Asn Pro Ser 50 5560

Leu Lys Ser Arg Val Thr Ile Ser Val Asp Thr Ser Lys Asn Gln Phe 65 70 7580

Ser Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr 85 9095

Cys Thr Arg Glu Asp Ser Ser Asn Trp Arg Ser Arg Gly Gln Gly Thr

100 105110

Leu Val Thr Val Ser Ser

115 <210> 70 <211> 118 <212> Protein <213> Artificial sequence <220>

<221> source

- 70 048216 <223> /note="Description of polypeptide" of artificial sequence

Synthetic <400> 70

Gln Leu Gln Leu 1

Gln Glu Ser Gly 5

Pro Gly Leu Val 10

Arg Pro Ser Glu 15

Th Leu Ser Leu

Thr Cys Thr Val

Ser Gly Gly Ser 25

Ile Ser Ser Ser 30

Ser Tyr Tyr Trp 35

Gly Trp Ile Arg 40

Gln Pro Pro Gly

Lys Gly Leu Glu 45

Trp Ile Gly Ser 50

Ile Tyr Tyr Ser 55

Gly Ser Thr Tyr

Tyr Asn Pro Ser

Leu Lys Ser Arg 65

Val Thr Ile Ser

Val Asp Thr Ser 75

Lys Asn Gln Phe

Sir Leu Lys Leu

Ser Ser Val Thr 85

Ala Ala Asp Thr 90

Ala Val Tyr Tyr

CysThrArgGlu

100

Asp Ser Ser Ser

Trp Arg Ser Arg 105

Gly Gln Gly Thr 110

Leu Val Thr Val Ser Ser

115 <210> 71 <211> 118 <212> Protein <213> Artificial sequence <220>

<221> source <223> /note="Description of Artificial Sequence: Synthetic Polypeptide" <400> 71

Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu

5 1015

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Gly Ser Ile Ser SerSer

2530

Ser Tyr Tyr Trp Gly Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu 35 4045

Trp Ile Gly Ser Ile Tyr Tyr Ser Gly Ala Thr Tyr Tyr Asn Pro Ser

5560

- 71 048216

Leu Lys Asn Arg Val Thr Ile Ser Val Asp Thr Ser Arg Asn Gln Phe 65 70 7580

Ser Leu Asn Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr 85 9095

Cys Thr Arg Glu Asp Ser Ser Asn Trp Arg Ser Arg Gly Gln Gly Thr

100 105110

Leu Val Thr Val Ser Ser

115 <210> 72 <211> 118 <212> Protein <213> Artificial sequence <220>

<221> source <223> /note="Description of Artificial Sequence: Synthetic Polypeptide" <400> 72

Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu

5 1015

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Gly Ser Ile Ser Ser Ser 20 2530

Ser Tyr Tyr Trp Gly Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu 35 4045

Trp Ile Gly Ser Ile Tyr Tyr Ser Gly Ala Thr Tyr Tyr Asn Pro Ser 50 5560

Leu Lys Asn Arg Val Thr Ile Ser Val Asp Thr Ser Arg Asn Gln Phe 65 70 7580

Ser Leu Asn Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Met Tyr Tyr 85 9095

Cys Thr Arg Glu Asp Ser Ser Asn Trp Arg Ser Arg Gly Gln Gly Thr

100 105110

Leu Val Thr Val Ser Ser

115 <210> 73 <211> 118 <212> Protein <213> Artificial sequence

- 72 048216 <220>

<221> source <223> /note="Description of Artificial Sequence: Synthetic Polypeptide" <400> 73

Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu

5 10 15

Th Leu Ser Leu

Thr Cys Thr Val

Ser Gly Gly Ser 25

Ile Ser Ser Ser 30

Ser Tyr Tyr Trp 35

Gly Trp Ile Arg 40

Gln Pro Pro Gly

Lys Gly Leu Glu 45

Trp Ile Gly Ser 50

Ile Tyr Tyr Ser 55

Gly Val Thr Tyr

Tyr Asn Pro Ser

Leu Lys Asn Arg 65

Val Thr Ile Ser

Val Asp Thr Ser 75

Lys Asn Gln Phe

Sir Leu Lys Leu

Ser Ser Val Thr 85

Ala Ala Asp Thr 90

Ala Val Tyr Tyr 95

CysThrArgGlu

100

Asp Ser Ser Ser

Trp Arg Ser Arg 105

Gly Gln Gly Thr 110

Leu Val Thr Val Ser Ser

115 <210> 74 <211> 118 <212> Protein <213> Artificial sequence <220>

<221> source <223> /note="Description of Artificial Sequence: Synthetic Polypeptide" <400> 74

Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu

5 1015

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Asp Ser Ile Ser SerGly

2530

Asp Tyr Tyr Trp Gly Trp Ile Arg Gln Ser Pro Glu Lys Gly Leu Glu

4045

Trp Ile Gly His Ile Tyr Tyr Ser Gly Val Thr Tyr Tyr Asn Pro Ser

- 73 048216

Leu Lys Asn Arg Val Thr Ile Ser Val Asp Thr Ser Lys Asn Gln Phe 65 70 7580

Ser Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr 85 9095

Cys Lys Arg Asp Asp Ser Ser Asn Trp Arg Ser Arg Gly Gln Gly Thr

100 105110

Leu Val Thr Val Ser Ser

115 <210> 75 <211> 118 <212> Protein <213> Artificial sequence <220>

<221> source <223> /note="Description of Artificial Sequence: Synthetic Polypeptide" <400> 75

Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu

5 1015

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Gly Ser Ile Ser Ser Ser 20 2530

Ser Tyr Tyr Trp Gly Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu 35 4045

Trp Ile Gly His Ile Tyr Tyr Ser Gly Ala Thr Tyr Tyr Asn Pro Ser 50 5560

Leu Lys Asn Arg Val Thr Ile Ser Val Asp Thr Ser Lys Asn Gln Phe 65 70 7580

Ser Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr 85 9095

Cys Thr Arg Glu Asp Ser Ser Ser Trp Arg Ser Arg Gly Gln Gly Thr

100 105110

Leu Val Thr Val Ser Ser

115 <210> 76

- 74 048216 <211> 118 <212> Protein <213> Artificial sequence <220>

<221> source <223> /note="Description of Artificial Sequence: Synthetic Polypeptide" <400> 76

Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu

5 1015

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Gly Ser Ile Ser SerSer

2530

Ser Tyr Tyr Trp Gly Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu 35 4045

Trp Ile Gly Ser Val Tyr Tyr Thr Gly Ala Thr Tyr Tyr Asn Pro Ser

5560

Leu Lys Asn Arg Val Thr Ile Ser Val Asp Thr Ser Lys Asn GlnPhe

70 7580

Ser Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val TyrTyr

9095

Cys Thr Arg Asp Asp Ser Ser Asn Trp Arg Ser Arg Gly Gln Gly Thr 100 105110

Leu Val Thr Val Ser Ser

115 <210> 77 <211> 118 <212> Protein <213> Artificial sequence <220>

<221> source <223> /note="Description of Artificial Sequence: Synthetic Polypeptide" <400> 77

Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu

5 1015

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Gly Ser Ile Ser SerSer

2530

Ser Tyr Tyr Trp Gly Trp Phe Arg His Pro Pro Gly Lys Gly Leu Asp

4045

- 75 048216

Trp Ile Gly Ser Ile His Tyr Ser Gly Ser Thr Tyr Tyr Asn Pro Ser 50 5560

Leu Lys Ser Arg Val Thr Ile Ser Val Asp Thr Ser Arg Asn Gln Phe 65 70 7580

Ser Leu Asn Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr 85 9095

Cys Thr Arg Asp Asp Ser Ser Asn Trp Arg Ser Arg Gly Gln Gly Thr

100 105110

Leu Val Thr Val Ser Ser

115 <210> 78 <211> 118 <212> Protein <213> Artificial sequence <220>

<221> source <223> /note="Description of Artificial Sequence: Synthetic Polypeptide" <400> 78

Gln Leu Gln Leu Gln Glu Ser Asp Pro Gly Leu Val Lys Pro Ser Glu

5 1015

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Gly Ser Ile Ser Ser Ser 20 2530

Ser His Tyr Trp Gly Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu 35 4045

Trp Ile Gly His Ile Tyr Tyr Ser Gly Ala Thr Tyr Tyr Asn Pro Ser 50 5560

Leu Lys Asn Arg Val Thr Ile Ser Val Asp Thr Ser Arg Asn Gln Phe 65 70 7580

Ser Leu Asn Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr 85 9095

Cys Thr Arg Glu Asp Ser Ser Asn Trp Arg Ser Arg Gly Gln Gly Thr

100 105110

Leu Val Thr Val Ser Ser

115

- 76 048216 <210> 79 <211> 118 <212> Protein <213> Artificial sequence <220>

<221> source <223> /note="Description of Artificial Sequence: Synthetic Polypeptide" <400> 79

Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu

5 1015

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Gly Ser Ile Ser Ser Ser 20 2530

Ser Tyr Tyr Trp Gly Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu 35 4045

Trp Ile Gly Ser Ile Tyr Tyr Ser Gly Ser Thr Tyr Tyr Asn Pro Ser 50 5560

Leu Lys Asn Arg Val Thr Ile Ser Val Asp Thr Ser Arg Asn Gln Phe 65 70 7580

Ser Leu Asn Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr 85 9095

Cys Thr Arg Asp Asp Ser Ser Asn Trp Arg Ser Arg Gly Gln Gly Thr

100 105110

Leu Val Thr Val Ser Ser

115 <210> 80 <211> 118 <212> Protein <213> Artificial sequence <220>

<221> source <223> /note="Description of Artificial Sequence: Synthetic Polypeptide" <400> 80

Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu

5 10 15

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Gly Ser Ile Ser Ser Ser

25 30

- 77 048216

Ser Tyr Tyr Trp Gly Trp Ile Arg

40

Gln Pro Pro Gly Lys Gly Leu Glu 45

Trp Ile Gly His Ile Tyr Tyr Ser

55

Gly Ala Thr Tyr Tyr Asn Pro Ser 60

Leu Lys Asn Arg Val Thr Ile Ser 65 70

Val Asp Thr Ser Arg Asn Gln Phe

80

Ser Leu Asn Leu Ser Ser Val Thr

Ala Ala Asp Thr Ala Val Tyr Tyr

95

Cys Thr Arg Glu Asp Ser Ser Asn

100

Trp Arg Ser Arg Gly Gln Gly Thr

105 110

Leu Val Thr Val Ser Ser

115 <210> 81 <211> 118 <212> Protein <213> Artificial sequence <220>

<221> source <223> /note="Description of Artificial Sequence: Synthetic Polypeptide" <400> 81

Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu

5 1015

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Gly Ser Ile Ile Ser Ser 20 2530

Ser Tyr Tyr Trp Gly Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu 35 4045

Trp Ile Gly Ser Ile Tyr Tyr Ser Gly Ser Ala Tyr Tyr His Pro Ser 50 5560

Leu Lys Ser Arg Val Thr Ile Ser Ile Asp Thr Ser Lys Asn Gln Phe 65 70 7580

Ser Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr 85 9095

Cys Ala Arg Asp Asp Ser Ser Asn Trp Arg Ser Arg Gly Gln Gly Thr

100 105110

- 78 048216

Leu Val Thr Val Ser Ser

115 <210> 82 <211> 118 <212> Protein <213> Artificial sequence <220>

<221> source <223> /note="Description of Artificial Sequence: Synthetic Polypeptide" <400> 82

Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu

5 1015

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Gly Ser Ile Ser Ser Ser 20 2530

Ser Tyr Tyr Trp Gly Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu 35 4045

Trp Ile Gly Ser Ile Tyr Tyr Ser Gly Ala Thr Tyr Tyr Asn Pro Ser 50 5560

Leu Lys Asn Arg Val Thr Ile Ser Val Asp Thr Ser Arg Asn Gln Phe 65 70 7580

Ser Leu Asn Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr 85 9095

Cys Thr Arg Asp Asp Ser Ser Asn Trp Arg Ser Arg Gly Gln Gly Thr

100 105110

Leu Val Thr Val Ser Ser

115 <210> 83 <211> 118 <212> Protein <213> Artificial sequence <220>

<221> source <223> /note="Description of Artificial Sequence: Synthetic Polypeptide" <400> 83

Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu

5 10 15

- 79 048216

Thr Leu Ser Leu Thr Cys Thr Val

Ser Gly Asp Ser Ile Ser Ser Gly 25 30

Asp Tyr Tyr Trp Gly Trp Ile Arg

40

Gln Pro Pro Gly Lys Gly Leu Glu 45

Trp Ile Gly His Ile Tyr Tyr Ser

55

Gly Ala Thr Tyr Tyr Asn Pro Ser 60

Leu Lys Asn Arg Val Thr Ile Ser 65 70

Val Asp Thr Ser Arg Asn Gln Ser

80

Ser Leu Asn Leu Ser Ser Val Thr

Ala Ala Asp Thr Ala Val Tyr Tyr

95

Cys Thr Arg Glu Asp Ser Ser Asn

100

Trp Arg Ser Arg Gly Gln Gly Thr

105 110

Leu Val Thr Val Ser Ser

115 <210> 84 <211> 118 <212> Protein <213> Artificial sequence <220>

<221> source <223> /note="Description of Artificial Sequence: Synthetic Polypeptide" <400> 84

Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu

5 1015

Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Gly Ser Ile Asn Asp Asn 20 2530

Ser His Tyr Trp Gly Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu 35 4045

Trp Ile Gly His Ile Tyr Tyr Ser Gly Ala Thr Tyr Tyr Asn Pro Ser 50 5560

Leu Lys Asn Arg Val Thr Ile Ser Val Asp Thr Ser Arg Asn Gln Phe 65 70 7580

Ser Leu Asn Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr 85 9095

- 80 048216

Cys Thr Arg Glu Asp Ser Ser Ser Trp Arg Ser Arg Gly Gln Gly Thr

100 105 110

Leu Val Thr Val Ser Ser

115 <210> 85 <211> 8 <212> Protein <213> Artificial sequence <220>

<221> source <223> /note="Description of the artificial sequence: Synthetic peptide" <400> 85

Gly Phe Thr Phe His Asn Tyr Ala

5 <210> 86 <211> 8 <212> Protein <213> Artificial sequence <220>

<221> source <223> /note="Description of artificial sequence: Synthetic peptide" <400> 86

Ile Ser Trp Asn Ser Gly Ser Ile

5 <210> 87 <211> 16 <212> Protein <213> Artificial sequence <220>

<221> source <223> /note="Description of artificial sequence: Synthetic peptide" <400> 87

Ala Lys Asp Ser Arg Gly Tyr Gly Asp Tyr Ser Leu Gly Gly Ala Tyr 1 5 10 15 <210> 88 <211> 6 <212> Protein <213> Artificial sequence <220>

<221> source <223> /note="Description of the synthetic sequence: Synthetic peptide"

- 81 048216 <400> 88

Gln Ser Val Ser Ser Asn <210> 89 <211> 3 <212> Protein <213> Artificial sequence <220>

<221> source <223> /note="Description of artificial sequence: Synthetic peptide" <400> 89

Gly Ala Ser <210> 90 <211> 9 <212> Protein <213> Artificial sequence <220>

<221> source <223> /note="Description of the artificial sequence: Synthetic peptide" <400> 90

Gln Gln Tyr Asn Asn Trp Pro Trp Thr

5 <210> 91 <211> 142 <212> Protein <213> Artificial sequence <220>

<221> source <223> /note="Description of Artificial Sequence: Synthetic Polypeptide" <400> 91

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Arg

5 1015

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe His Asn Tyr 20 2530

Ala Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 4045

Ser Gly Ile Ser Trp Asn Ser Gly Ser Ile Gly Tyr Ala Asp Ser Val 50 5560

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr

- 82 048216

70 7580

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Leu Tyr Tyr Cys 85 9095

Ala Lys Asp Ser Arg Gly Tyr Gly Asp Tyr Ser Leu Gly Gly Ala Tyr 100 105110

Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Asp Tyr Arg Leu Gly 115 120125

Gly Ala Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser 130 135140 <210> 92 <211> 107 <212> Protein <213> Artificial sequence <220>

<221> source <223> /note="Description of Artificial Sequence: Synthetic Polypeptide" <400> 92

Glu Ile Val Met Thr Gln Ser Pro Ala Thr Leu Ser Val Ser Pro Gly

5 1015

Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Asn 20 2530

Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile 35 4045

Tyr Gly Ala Ser Thr Arg Ala Thr Gly Ile Pro Ala Arg Phe Ser Gly 50 5560

Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Ser

70 7580

Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr Asn Asn Trp Pro Trp 85 9095

Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys

100105 <210> 93 <211> 330 <212> Protein <213> Homo sapiens

- 83 048216 <400> 93

Ala Ser Thr Lys Gly 1 5

Pro Ser Val Phe Pro

Leu Ala Pro Ser Ser

Ser Thr Ser Gly Gly

Thr Ala Ala Leu Gly

Cys Leu Val Lys Asp

Phe Pro Glu Pro Val

Thr Val Ser Trp Asn 40

Ser Gly Ala Leu Thr 45

Gly Val His Thr Phe 50

About Ala Val Leu Gln

Ser Ser Gly Leu Tyr 60

Leu Ser Ser Val Val 65

Thr Val Pro Ser Ser 70

Ser Leu Gly Thr Gln 75

Tyr Ile Cys Asn Val

Asn His Lys Pro Ser

Asn Thr Lys Val Asp

Lys Val Glu Pro Lys

100

Ser Cys Asp Lys Thr

105

His Thr Cys Pro Pro

110

Pro Ala Pro Glu Leu

115

Leu Gly Gly Pro Ser

120

Val Phe Leu Phe Pro

125

Lys Pro Lys Asp Th

130

Leu Met Ile Ser Arg

135

Thr Pro Glu Val Thr

140

Val Val Val Asp Val 145

Ser His Glu Asp Pro 150

Glu Val Lys Phe Asn 155

Tyr Val Asp Gly Val

165

Glu Val His Asn Ala

170

Lys Thr Lys Pro Arg

175

Glu Gln Tyr Asn Ser

180

Thr Tyr Arg Val Val

185

Ser Val Leu Thr Val

190

His Gln Asp Trp Leu 195

Asn Gly Lys Glu Tyr

200

Lys Cys Lys Val Ser 205

Lys Ala Leu Pro Ala

210

Pro Ile Glu Lys Thr 215

Ile Ser Lys Ala Lys

220

Gln Pro Arg Glu Pro 225

Gln Val Tyr Thr Leu 230

Pro Pro Ser Arg Asp 235

Leu Thr Lys Asn Gln

245

Val Ser Leu Thr Cys

250

Leu Val Lys Gly Phe

255

Lys

Tyr

Ser

Ser

Th 80

Lys

Cys

Pro

Cys

Trp 160

Glu

Leu

Asn

Gly

Glu 240

Tyr

- 84 048216

Pro Ser Asp Ile Ala

260

Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn

265 270

Asn Tyr Lys Thr Thr 275

Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

280 285

Leu Tyr Ser Lys Leu 290

Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn

295 300

Val Phe Ser Cys Ser 305

Val Met His Glu Ala Leu His Asn His Tyr Thr

310 315 320

Gln Lys Ser Leu Ser

325

Leu Ser Pro Gly Lys

330 <210> 94 <211> 327 <212> Protein <213> Homo sapiens <400> 94

Ala Ser Thr Lys Gly 1 5

Pro Ser Val Phe Pro Leu Ala Pro Cys Ser Arg

15

Ser Thr Ser Glu Ser

Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr

30

Phe Pro Glu Pro Val

Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser

45

Gly Val His Thr Phe 50

Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser

60

Leu Ser Ser Val Val 65

Thr Val Pro Ser Ser Ser Leu Gly Thr Lys Thr 70 75 80

Tyr Thr Cys Asn Val

Asp His Lys Pro Ser Asn Thr Lys Val Asp Lys 90 95

Arg Val Glu Ser Lys

100

Tyr Gly Pro Pro Cys Pro Ser Cys Pro Ala Pro

105 110

Glu Phe Leu Gly Gly 115

Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys

120 125

Ser Arg Thr Pro Glu Val Thr Cys Val Val Val

135 140

Asp Thr Leu Met Ile

130

- 85 048216

Asp Val Ser Gln Glu Asp Pro Glu

145 150

Val Gln Phe Asn Trp Tyr Val Asp

155 160

Gly Val Glu Val

His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe

165 170 175

Asn Ser Thr Tyr Arg Val Val

180

Ser Val Leu Thr Val Leu His Gln Asp

185 190

Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val

195 200

Ser Asn Lys Gly Leu

205

Pro Ser Ser Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg

210 215 220

Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Gln Glu Glu Met Thr Lys

225 230 235 240

Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp

245 250 255

Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys

260 265 270

Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser

275 280 285

Arg Leu Thr Val Asp Lys Ser Arg Trp Gln Glu Gly Asn Val Phe Ser

290 295 300

Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser

305 310 315 320

Leu Ser Leu Ser Leu Gly Lys 325 <210> 95 <211> 330 <212> Protein <213> Artificial sequence <220>

<221> source <223> /note="Description of Artificial Sequence: Synthetic Polypeptide" <400> 95

Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys

5 10 15

- 86 048216

Ser Thr Ser Gly Gly

Thr Ala Ala Leu Gly

Cys Leu Val Lys Asp

Phe Pro Glu Pro Val

Thr Val Ser Trp Asn 40

Ser Gly Ala Leu Thr 45

Gly Val His Thr Phe 50

About Ala Val Leu Gln

Ser Ser Gly Leu Tyr 60

Leu Ser Ser Val Val 65

Thr Val Pro Ser Ser 70

Ser Leu Gly Thr Gln 75

Tyr Ile Cys Asn Val

Asn His Lys Pro Ser

Asn Thr Lys Val Asp

Lys Val Glu Pro Lys

100

Ser Cys Asp Lys Thr

105

His Thr Cys Pro Pro

110

Pro Ala Pro Glu Ala

115

Ala Gly Gly Pro Ser 120

Val Phe Leu Phe Pro

125

Lys Pro Lys Asp Th

130

Leu Met Ile Ser Arg

135

Thr Pro Glu Val Thr

140

Val Val Val Asp Val 145

Ser His Glu Asp Pro 150

Glu Val Lys Phe Asn 155

Tyr Val Asp Gly Val

165

Glu Val His Asn Ala

170

Lys Thr Lys Pro Arg

175

Glu Gln Tyr Asn Ser

180

Thr Tyr Arg Val Val

185

Ser Val Leu Thr Val

190

His Gln Asp Trp Leu 195

AsnGlyLysGluTyr200

Lys Cys Lys Val Ser 205

Lys Ala Leu Pro Ala

210

Pro Ile Glu Lys Thr 215

Ile Ser Lys Ala Lys

220

Gln Pro Arg Glu Pro 225

Gln Val Tyr Thr Leu 230

Pro Pro Ser Arg Glu 235

Met Thr Lys Asn Gln

245

Val Ser Leu Thr Cys

250

Leu Val Lys Gly Phe

255

Pro Ser Asp Ile Ala

260

Val Glu Trp Glu Ser

265

Asn Gly Gln Pro Glu

270

Tyr

Ser

Ser

Th 80

Lys

Cys

Pro

Cys

Trp 160

Glu

Leu

Asn

Gly

Glu 240

Tyr

Asn

- 87 048216

Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe

275 280285

Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn

290 295300

Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr

305 310 315320

Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys

325330 <210> 96 <211> 327 <212> Protein <213> Artificial sequence <220>

<221> source <223> /note="Description of Artificial Sequence: Synthetic Polypeptide" <400> 96

Ala Ser Thr Lys Gly Pro Ser Val

5

Phe Pro Leu Ala Pro Cys Ser Arg

15

Ser Thr Ser Glu Ser Thr Ala Ala

Leu Gly Cys Leu Val Lys Asp Tyr 25 30

Phe Pro Glu Pro Val Thr Val Ser

40

Trp Asn Ser Gly Ala Leu Thr Ser 45

Gly Val His Thr Phe Pro Ala Val

55

Leu Gln Ser Ser Gly Leu Tyr Ser 60

Leu Ser Ser Val Val Thr Val Pro

70

Ser Ser Ser Leu Gly Thr Lys Thr

80

Tyr Thr Cys Asn Val Asp His Lys 85

Pro Ser Asn Thr Lys Val Asp Lys

95

Arg Val Glu Ser Lys Tyr Gly Pro

100

Pro Cys Pro Pro Cys Pro Ala Pro

105 110

Glu Ala Ala Gly Gly Pro Ser Val

115 120

Phe Leu Phe Pro Pro Lys Pro Lys

125

Asp Thr Leu Met Ile Ser Arg Thr

130 135

Pro Glu Val Thr Cys Val Val Val

140

- 88 048216

Asp Val Ser Gln Glu Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp

145 150 155160

Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe

165 170175

Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp

180 185190

Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu

195 200205

Pro Ser Ser Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg

210 215220

Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Gln Glu Glu Met Thr Lys

225 230 235240

Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp

245 250255

Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys

260 265270

Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser

275 280285

Arg Leu Thr Val Asp Lys Ser Arg Trp Gln Glu Gly Asn Val Phe Ser

290 295300

Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser

305 310 315320

Leu Ser Leu Ser Leu Gly Lys

325 <210> 97 <211> 214 <212> Protein <213> Artificial sequence <220>

<221> source <223> /note="Description of Artificial Sequence: Synthetic Polypeptide" <400> 97

Glu Ile Val Met Thr Gln Ser Pro Ala Thr Leu Ser Val Ser Pro Gly

5 10 15

- 89 048216

Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln

25

Ser Val Ser Ser Asn

Leu Ala

Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu 35 40 45

Ile

Tyr Gly Ala Ser Thr Arg Ala Thr Gly Ile Pro Ala Arg Phe Ser Gly 50 55 60

Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Ser 65 70 75 80

Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr Asn Asn Trp Pro Trp 85 90 95

Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys Arg Thr Val Ala Ala

100 105 110

Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly

115 120 125

Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala

130 135 140

Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln

145 150 155 160

Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser

165 170 175

Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr

180 185 190

Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser

195 200 205

Phe Asn Arg Gly Glu Cys

210 <210> 98 <211> 453 <212> Protein <213> Artificial sequence <220>

<221> source <223> /note="Description of Artificial Sequence: Synthetic Polypeptide"

- 90 048216 <400> 98

Glu Val Gln Leu Val

5

Glu Ser Gly Gly Gly

Leu Val Gln Pro Gly

Ser Leu Arg Leu Ser

Cys Ala Ala Ser Gly

Phe Thr Phe His Asn

Ala Met His Trp Val 35

Arg Gln Ala Pro Gly 40

LysGlyLeuGluTrp45

Ser Gly Ile Ser Trp 50

Asn Ser Gly Ser Ile 55

Gly Tyr Ala Asp Ser 60

LysGlyArgPheThr65

Ile Ser Arg Asp Asn 70

Ala Lys Asn Ser Leu 75

Leu Gln Met Asn Ser

Leu Arg Ala Glu Asp

Thr Ala Leu Tyr Tyr

Ala Lys Asp Ser Arg

100

Gly Tyr Gly Asp Tyr

105

Ser Leu Gly Gly Ala

110

TrpGlyGlnGlyThr115

Leu Val Thr Val Ser

120

Ser Ala Ser Thr Lys

125

Pro Ser Val Phe Pro

130

Leu Ala Pro Ser Ser

135

LysSerThrSerGly140

Thr Ala Ala Leu Gly 145

Cys Leu Val Lys Asp 150

Tyr Phe Pro Glu Pro 155

Thr Val Ser Trp Asn

165

Ser Gly Ala Leu Thr

170

Ser Gly Val His Thr

175

About Ala Val Leu Gln

180

Ser Ser Gly Leu Tyr

185

Sir Leu Sir Sir Val

190

Thr Val Pro Ser Ser

195

Ser Leu Gly Thr Gln

200

Thr Tyr Ile Cys Asn 205

Asn His Lys Pro Ser 210

Asn Thr Lys Val Asp

215

Lys Lys Val Glu Pro

220

Ser Cys Asp Lys Thr 225

His Thr Cys Pro Pro 230

Cys Pro Ala Pro Glu 235

Leu Gly Gly Pro Ser

Val Phe Leu Phe Pro

Pro Lys Pro Lys Asp

Arg

Tyr

Val

Val

Tyr 80

Cys

Tyr

Gly

Gly

Val 160

Phe

Val

Val

Lys

Leu 240

Th

- 91 048216

245 250255

Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val

260 265270

Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val

275 280285

Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser

290 295300

Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu

305 310 315320

Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala

325 330335

Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro

340 345350

Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln

355 360365

Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala

370 375380

Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr

385 390 395400

Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu

405 410415

Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser

420 425430

Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser

435 440445

Leu Ser Pro Gly Lys 450 <210> 99 <211> 453 <212> Protein <213> Artificial sequence <220>

<221> source <223> /note="Description of the artificial sequence: Synthetic

- 92 048216 polypeptide" <400> 99

Glu Val Gln Leu Val 1 5

Ser Leu Arg Leu Ser

Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Arg 10 15

Cys Ala Ala Ser Gly Phe Thr Phe His Asn Tyr 25 30

Ala Met His Trp Val 35

Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

45

Ser Gly Ile Ser Trp 50

Asn Ser Gly Ser Ile Gly Tyr Ala Asp Ser Val

60

LysGlyArgPheThr65

Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr

75 80

Leu Gln Met Asn Ser

Leu Arg Ala Glu Asp Thr Ala Leu Tyr Tyr Cys

95

Ala Lys Asp Ser Arg

100

Gly Tyr Gly Asp Tyr Ser Leu Gly Gly Ala Tyr

105 110

TrpGlyGlnGlyThr115

Leu Val Thr Val

120

Ser Ser Ala Ser Thr Lys Gly

125

Pro Ser Val Phe Pro

130

Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly

135 140

Thr Ala Ala Leu Gly 145

Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val

150 155 160

Thr Val Ser Trp Asn

165

Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe

170 175

About Ala Val Leu Gln

180

Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val

185 190

Thr Val Pro Ser Ser

195

Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val

200 205

Asn His Lys Pro Ser 210

Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys

215 220

Ser Cys Asp Lys Thr 225

His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala

230 235 240

- 93 048216

Ala Gly Gly Pro Ser Val Phe Leu Phe Pro Pro

245250

Lys Pro Lys Asp Th

255

Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val

260 265270

Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val

275 280285

Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser

290 295300

Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu

305 310 315320

Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala

325 330335

Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro

340 345350

Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln

355 360365

Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala

370 375380

Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr

385 390 395400

Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu

405 410415

Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser

420 425430

Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser

435 440445

Leu Ser Pro Gly Lys

450 <210> 100 <211> 450 <212> Protein <213> Artificial sequence <220>

<221> source

- 94 048216 <223> /note="Description of artificial sequence: Synthetic polypeptide" <400> 100

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Arg

5 1015

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe His Asn Tyr 20 2530

Ala Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 4045

Ser Gly Ile Ser Trp Asn Ser Gly Ser Ile Gly Tyr Ala Asp Ser Val 50 5560

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr 65 70 7580

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Leu Tyr Tyr Cys 85 9095

Ala Lys Asp Ser Arg Gly Tyr Gly Asp Tyr Ser Leu Gly Gly Ala Tyr

100 105110

Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly

115 120125

Pro Ser Val Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser

130 135140

Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val

145 150 155160

Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe

165 170175

Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val

180 185190

Thr Val Pro Ser Ser Ser Leu Gly Thr Lys Thr Tyr Thr Cys Asn Val

195 200205

Asp His Lys Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu Ser Lys

210 215220

Tyr Gly Pro Pro Cys Pro Ser Cys Pro Ala Pro Glu Phe Leu Gly Gly

225 230 235240

- 95 048216

Pro Ser Val Phe Leu

245

Phe Pro Pro Lys Pro Lys Asp Thr Leu Met

250 255

Ser Arg Thr Pro Glu

260

Val Thr Cys Val Val Val Asp Val Ser Gln

265 270

Asp Pro Glu Val Gln

275

Phe Asn Trp Tyr Val Asp Gly Val Glu Val

280 285

Asn Ala Lys Thr Lys

290

Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr

295 300

Val Val Ser Val Leu 305

Thr Val Leu His Gln Asp Trp Leu Asn Gly

310 315

Glu Tyr Lys Cys Lys

325

Val Ser Asn Lys Gly Leu Pro Ser Ser Ile

330 335

Lys Thr Ile Ser Lys

340

Ala Lys Gly Gln Pro Arg Glu Pro Gln Val

345 350

Thr Leu Pro Pro Ser

355

Gln Glu Glu Met Thr Lys Asn Gln Val Ser

360 365

Thr Cys Leu Val Lys

370

Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu

375 380

Glu Ser Asn Gly Gln 385

Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro

390 395

Leu Asp Ser Asp Gly

405

Ser Phe Phe Leu Tyr Ser Arg Leu Thr Val

410 415

Lys Ser Arg Trp Gln

420

Glu Gly Asn Val Phe Ser Cys Ser Val Met

425 430

Glu Ala Leu His Asn

435

His Tyr Thr Gln Lys Ser Leu Ser Leu Ser

440 445

Ile

Glu

His

Arg

Lys 320

Glu

Tyr

Leu

Trp

Val 400

Asp

His

Leu

Gly Lys

450 <210> 101 <211> 450 <212> Protein <213> Artificial <220>

subsequence

- 96 048216 <221> source <223> /note="Description of Artificial Sequence: Synthetic Polypeptide" <400> 101

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Arg

5 1015

Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe His AsnTyr

2530

Ala Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 4045

Ser Gly Ile Ser Trp Asn Ser Gly Ser Ile Gly Tyr Ala Asp Ser Val

5560

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr 65 70 7580

Leu Gln Met Asn

Ser Leu Arg Ala 85

Glu Asp Thr Ala 90

Leu Tyr Tyr Cys 95

Ala Lys Asp Ser

100

ArgGlyTyrGly

Asp Tyr Ser Leu 105

Gly Gly Ala Tyr

110

TrpGlyGlnGly

115

Thr Leu Val Thr

120

Val Ser Ser Ala

Ser Thr Lys Gly 125

Pro Ser Val Phe Pro Leu Ala Pro Cys

130 135

Ser Arg Ser Thr Ser Glu Ser 140

Thr Ala Ala Leu 145

Gly Cys Leu Val 150

Lys Asp Tyr Phe

155

Pro Glu Pro Val

160

Thr Val Ser Trp

Asn Ser Gly Ala 165

Leu Thr Ser Gly 170

Val His Thr Phe

175

About Ala Val Leu

180

Gln Ser Ser Gly

Leu Tyr Ser Leu 185

Ser Ser Val Val

190

Thr Val Pro Ser

195

Ser Ser Leu Gly

200

Thr Lys Thr Tyr

Thr Cys Asn Val 205

Asp His Lys Pro 210

Ser Asn Thr Lys

215

Val Asp Lys Arg

220

Val Glu Ser Lys

Tyr Gly Pro Pro Cys Pro Pro Cys

225 230

About Ala

Pro Glu Ala Ala Gly Gly

235 240

- 97 048216

Pro Ser Val Phe Leu

245

Phe Pro Pro Lys Pro Lys Asp Thr Leu Met

250 255

Ser Arg Thr Pro Glu

260

Val Thr Cys Val Val Val Asp Val Ser Gln

265 270

Asp Pro Glu Val Gln 275

Phe Asn Trp Tyr Val Asp Gly Val Glu Val

280 285

Asn Ala Lys Thr Lys

290

Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr

295 300

Val Val Ser Val Leu 305

Thr Val Leu His Gln Asp Trp Leu Asn Gly

310 315

Glu Tyr Lys Cys Lys

325

Val Ser Asn Lys Gly Leu Pro Ser Ser Ile

330 335

Lys Thr Ile Ser Lys

340

Ala Lys Gly Gln Pro Arg Glu Pro Gln Val

345 350

Thr Leu Pro Pro Ser

355

Gln Glu Glu Met Thr Lys Asn Gln Val Ser

360 365

Thr Cys Leu Val Lys

370

Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu

375 380

Glu Ser Asn Gly Gln 385

Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro

390 395

Leu Asp Ser Asp Gly

405

Ser Phe Phe Leu Tyr Ser Arg Leu Thr Val

410 415

Lys Ser Arg Trp Gln

420

Glu Gly Asn Val Phe Ser Cys Ser Val Met

425 430

Glu Ala Leu His Asn

435

His Tyr Thr Gln Lys Ser Leu Ser Leu Ser

440 445

Ile

Glu

His

Arg

Lys 320

Glu

Tyr

Leu

Trp

Val 400

Asp

His

Leu

Gly Lys

450 <210> 102 <211> 5 <212> Protein <213> Artificial sequence

- 98 048216 <220>

<221> source <223> /note="Description of artificial sequence: peptide" <400> 102

Gly Gly Gly Gly Ser

5

Synthetic <210> 103 <211> 10 <212> Protein <213> Artificial sequence <220>

<221> source <223> /note="Description of artificial sequence: peptide" <400> 103

Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser

5 10

Synthetic <210> 104 <211> 10 <212> Protein <213> Artificial sequence <220>

<221> source <223> /note="Description of the artificial sequence:

Synthetic peptide" <220>

<221> OPTION <222> (2) .. (2) <223> /33Μ6ημτβ=Ο17 <220>

<221> OPTION <222> (5)..(5) <223> /replace=TKg or Ile, or Asn <220>

<221> OPTION <222> (6)..(6) <223> /replace=Asp <220>

<221> OPTION <222> (7)..(7) <223> /replace='^ег or Asn <220>

<221> OPTION <222> (8) .. (8) <223> /replace=Gly or Ser <220>

<221> OPTION

- 99 048216 <222> (9)..(9) <223> /replace=Н1з <220>

<221> SITE <222> (1)..(10) <223> /note=Variant residues listed in the sequence are not preferred over those listed in the footnotes for variant positions <400> 104

Gly Asp Ser Ile Ser Ser Gly Asp Tyr Tyr

5 10 <210> 105 <211> 7 <212> Protein <213> Artificial sequence <220>

<221> source <223> /note="Description of the artificial sequence: Synthetic peptide" <220>

<221> OPTION <222> (1)..(1) <223> /replace=Val <220>

<221> OPTION <222> (2)..(2) <223> /replace=His <220>

<221> OPTION <222> (4)..(4) <223> /replace^'^^' <220>

<221> OPTION <222> (6)..(6) <223> /replace=Уа1 or Ser <220>

<221> OPTION <222> (7)..(7) <223> /replace=А1а <220>

<221> SITE <222> (1)..(7) <223> /note=Variant residues listed in the sequence are not preferred over those listed in the footnotes for variant positions <400> 105

Ile Tyr Tyr Ser Gly Ala Thr

5

- 100 048216 <210> 106 <211> 10 <212> Protein <213> Artificial sequence <220>

<221> source <223> /note="Description of the artificial sequence: Synthetic peptide" <220>

<221> OPTION <222> (1) .. (1) <223> /replace=A1a or Lys <220>

<221> OPTION <222> (3)..(3) <223> /33Μ6ημτβ=Ο1π ¹ '<220>

<221> OPTION <222> (7)..(7) <223> /replace=Ser <220>

<221> SITE <222> (1)..(10) <223> /note=Variant residues listed in the sequence are not preferred over those listed in the footnotes for variant positions <400> 106

Thr Arg Asp Asp Ser Ser Asn Trp Arg Ser

5 10 <210> 107 <211> 50 <212> Protein <213> Artificial sequence <220>

<221> source <223> /note="Description of Artificial Sequence: Synthetic Polypeptide" <220>

<221> SITE <222> (1)..(50) <223> /note="This sequence may span 1-10 repeat units "G1y G1y units "G1y G1y G1y G1y Ser" <220>

<221> source <223> /note="See the description as filed for a detailed description of substitutions and preferred embodiments"

- 101 -

Claims (12)

<400> 107 Gly 1 Gly Gly Gly Ser Gly Gly Gly Gly Ser 10 Gly Gly Gly Gly Ser 15 Gly Gly Gly Gly Ser 20 Gly Gly Gly Gly Ser 25 Gly Gly Gly Gly Ser 30 Gly Gly Gly Gly Ser 35 Gly Gly Gly Gly Ser 40 Gly Gly Gly Gly Ser 45 Gly Gly Gly Gly Ser CLAUSE OF INVENTION 1. A multispecific antibody comprising a first binding unit having binding affinity for CD22 and a second binding unit having binding affinity for CD3, wherein the first binding unit comprises a variable region comprising: (a) a CDR1 sequence according to SEQ ID NO: 1, a CDR2 sequence according to SEQ ID NO: 11 and a CDR3 sequence according to SEQ ID NO: 18; (b) a CDR1 sequence according to SEQ ID NO: 1, a CDR2 sequence according to SEQ ID NO: 12 and a CDR3 sequence according to SEQ ID NO: 19; or c) a CDR1 sequence according to SEQ ID NO: 1, a CDR2 sequence according to SEQ ID NO: 12 and a CDR3 sequence according to SEQ ID NO: 20; and wherein the second binding unit comprises: (a) a heavy chain variable region comprising: a CDR1 sequence according to SEQ ID NO: 85, a CDR2 sequence according to SEQ ID NO: 86 and a CDR3 sequence according to SEQ ID NO: 87; and (b) a light chain variable region comprising a CDR1 sequence according to SEQ ID NO: 88, a CDR2 sequence according to SEQ ID NO: 89 and a CDR3 sequence according to SEQ ID NO: 90. 2. The multispecific antibody of claim 1, wherein the first binding unit comprises a heavy chain variable region having at least 95% sequence identity to SEQ ID NO: 25. 3. The polyspecific antibody according to claim 1, wherein the first binding unit comprises the variable region of the heavy chain of the sequence SEQ ID NO: 25. 4. The polyspecific antibody of claim 1, wherein said CDR1, CDR2 and CDR3 sequences are present in a human VH framework. 5. A polyspecific antibody according to any one of claims 1 to 4, which is bispecific. 6. A polyspecific antibody containing: (i) a heavy chain variable region having binding affinity for CD3, comprising a CDR1 sequence according to SEQ ID NO: 85, a CDR2 sequence according to SEQ ID NO: 86 and a CDR3 sequence according to SEQ ID NO: 87 in a human VH framework; (ii) a light chain variable region comprising a CDR1 sequence according to SEQ ID NO: 88, a CDR2 sequence according to SEQ ID NO: 89 and a CDR3 sequence according to SEQ ID NO: 90 in a human VL framework; and (iii) an antigen-binding domain of an anti-CD22 heavy chain-only antibody comprising a CDR1 sequence according to SEQ ID NO: 1, a CDR2 sequence according to SEQ ID NO: 12 and a CDR3 sequence according to SEQ ID NO: 19 in a human VH framework. 7. A polyspecific antibody according to any one of claims 1 to 6, comprising the Fc region of human IgG1. 8. The polyspecific antibody of claim 7, wherein the human IgG1 Fc region is a silenced human IgG1 Fc region. 9. A polyspecific antibody according to any one of claims 1 to 6, comprising the Fc region of human IgG4. 10. The polyspecific antibody of claim 9, wherein the Fc region of human IgG4 comprises a mutation in the hinge region. 11. The polyspecific antibody of claim 9, wherein the human IgG4 Fc region comprises multiple knob-into-hole mutations. 12. The polyspecific antibody according to claim 9, wherein the Fc region of human IgG4 comprises a C-terminal lysine. - 102 -