JP5035522B2 - Vinyl polymer, blocking agent, and method for producing probe binding particles using the same - Google Patents

Description

  The present invention relates to a novel vinyl polymer, for example, a blocking agent applicable to the surface of immunodiagnostic particles, as well as probe-binding particles and a method for producing the same.

  In recent years, for the purpose of early detection of diseases, etc., it has been required to increase the sensitivity of tests, and improving the sensitivity of diagnostic agents has become a major issue. In diagnostic agents using a solid phase such as magnetic particles, in order to improve sensitivity, a method using enzyme color development as a detection method is being switched to a method using fluorescence or chemiluminescence that can obtain higher sensitivity. With the development of these detection techniques, it is theoretically said that it has reached a level at which even the presence of a single molecule of a test substance can be detected, but in reality, sufficient sensitivity has not been obtained. The reason for this is that in the diagnosis of detecting specific substances in the presence of biomolecules such as serum, coexisting biomolecules, secondary antibodies, luminescent substrates, etc. adsorb non-specifically to the solid phase, resulting in noise. Increasing the sensitivity is hindering. For this reason, in immunodiagnostic measurement, albumin, casein, gelatin are usually used to reduce the decrease in sensitivity due to non-specific adsorption of substances other than substances that specifically bind to the solid surface used for the immune reaction. By using a biological material such as a blocking agent, non-specific adsorption is suppressed and noise is reduced.

  However, even if the blocking operation by the conventional method is performed, non-specific adsorption still remains, and when a biologically derived blocking agent is used, there is a problem of biological contamination represented by BSE. Development of a high-performance blocking agent is desired.

As a blocking agent by chemical synthesis, polymers having polyoxyethylene have been proposed in JP-A-11-287802 and Japanese Patent No. 34079797, but the noise reduction effect by these blocking agents was insufficient. Furthermore, although a polyoxyethylene / pentaethylenehexamine block copolymer has been proposed as a blocking agent (Japanese Patent Laid-Open No. 2006-226882, Republished Patent No. WO 2005/010529), an industrially available polyoxy Ethylene often contains impurities having absorption in the vicinity of 290 nm, and complicated purification is required.
JP 11-287802 A Japanese Patent No. 34079797 JP 2006-226882 A Republished Patent No. WO2005 / 010529

  An object of the present invention is to provide a novel chemically synthesized vinyl polymer that is easy to manufacture and has sufficient noise reduction effect and signal enhancement effect in immunodiagnosis and the like, a blocking agent, and a method for producing probe-bound particles using the same. Is to provide.

  In order to solve this problem, the present inventors have found a vinyl polymer having a specific group and a blocking agent, and further, by treating the immunodiagnostic particle with the blocking agent, the particle can be obtained. It was found that a signal enhancement effect was exhibited, and the present invention was completed.

  The vinyl polymer which concerns on 1 aspect of this invention has group represented by following General formula (1).

・・・・・（１）
［式中、Ｒ^１およびＲ^２は独立に、水素原子、または、アミノ基およびイミノ基あるいはいずれか一方を１〜１１個有する基を表す（ただし、Ｒ^１およびＲ^２に含まれるアミノ基およびイミノ基の合計は２〜１１個である。）］
上記ビニルポリマーでは、上記一般式（１）において、Ｒ^１およびＲ^２がそれぞれ下記一般式（２ａ）および（２ｂ）で表される基であることができる。

(1)
[Wherein, R ¹ and R ² independently represent a hydrogen atom, an amino group and an imino group, or a group having 1 to 11 of either one (provided that the amino group contained in R ¹ and R ² and The total number of imino groups is 2 to 11.)]
In the vinyl polymer, in the general formula (1), R ¹ and R ² can be groups represented by the following general formulas (2a) and (2b), respectively.

・・・・・（２ａ）

(2a)

・・・・・（２ｂ）
［式中、ｘ１およびｘ２はそれぞれ１〜４で表される数であり、ｙ１およびｙ２はそれぞれ０〜１１で表される数である（ただし、ｙ１＋ｙ２は２〜１１を満たす。）］
上記ビニルポリマーは水溶性であることができる。

(2b)
[Wherein, x1 and x2 are each a number represented by 1 to 4, and y1 and y2 are each a number represented by 0 to 11 (provided that y1 + y2 satisfies 2 to 11)]
The vinyl polymer can be water soluble.

The vinyl polymer can be obtained by reacting a vinyl polymer polymerized in the presence of a compound having at least one group selected from the group consisting of a carboxyl group, an amino group and a hydroxyl group and a mercapto group with an oligoimine. In this case, the oligoimine may be a compound represented by the following general formula (4).
H ₂ N (CH ₂ CH ₂ NH) _n CH ₂ CH ₂ NH ₂ (4)
[In formula, n shows the number of 1-10. ]
The blocking agent which concerns on 1 aspect of this invention consists of the said vinyl polymer.

A method for producing probe-bound particles according to an aspect of the present invention includes:
Binding a probe to the surface of the particle;
Treating the particles to which the probe is bound using the blocking agent;
including.

  In the method for producing probe-bonded particles, in the step of bonding the probes, the particles may have at least one group selected from an active ester group, a tosyl group, and an epoxy group.

  In the method for producing probe-bonded particles, the particles may be magnetic particles.

  The method for producing probe-bound particles according to one embodiment of the present invention is obtained by the method for producing probe-bound particles.

  The probe binding particle according to one embodiment of the present invention has the blocking agent on the surface.

  The vinyl polymer and the blocking agent are easy to produce and have no possibility of biological contamination because they are chemically synthesized products, and have a higher noise reduction effect than the conventionally used blocking agents.

  Moreover, the said blocking agent can express a signal enhancement effect, when it uses for the particle | grains (for example, magnetic particle) used for an immunodiagnosis.

  Hereinafter, a vinyl polymer, a blocking agent, probe binding particles, and a production method thereof according to an embodiment of the present invention will be described.

1. Vinyl polymer having specific group and blocking agent, probe-binding particle and production method thereof 1.1. Configuration of Vinyl Polymer Having Specific Group (Blocking Agent) The vinyl polymer according to an embodiment of the present invention (also referred to as “modified vinyl polymer” in the present specification) is a group represented by the following general formula (1). Have For example, the vinyl polymer (terminal-modified vinyl polymer) according to the present embodiment can have a group represented by the following general formula (1) at the terminal.

・・・・・（１）
［式中、Ｒ^１およびＲ^２は独立に、水素原子、または、アミノ基およびイミノ基あるいはいずれか一方を１〜１１個有する基を表す（ただし、Ｒ^１およびＲ^２に含まれるアミノ基およびイミノ基の合計は２〜１１個である。）］
上記一般式（１）において、Ｒ^１およびＲ^２がそれぞれ下記一般式（２ａ）および（２ｂ）で表される基であってもよい。

(1)
[Wherein, R ¹ and R ² independently represent a hydrogen atom, an amino group and an imino group, or a group having 1 to 11 of either one (provided that the amino group contained in R ¹ and R ² and The total number of imino groups is 2 to 11.)]
In the general formula (1), R ¹ and R ² may be groups represented by the following general formulas (2a) and (2b), respectively.

・・・・・（２ａ）

(2a)

・・・・・（２ｂ）
［式中、ｘ１およびｘ２はそれぞれ１〜４で表される数であり、ｙ１およびｙ２はそれぞれ０〜１１で表される数である（ただし、ｙ１＋ｙ２は２〜１１を満たす。）］
上記一般式（２ａ）および（２ｂ）において、ｘ１およびｘ２は２であるのが好ましく、ｙ１およびｙ２はそれぞれ１〜６（ただし、ｙ１＋ｙ２＝２〜１１を満たす。）であることがより好ましい。

(2b)
[Wherein, x1 and x2 are each a number represented by 1 to 4, and y1 and y2 are each a number represented by 0 to 11 (provided that y1 + y2 satisfies 2 to 11)]
In the general formulas (2a) and (2b), x1 and x2 are preferably 2, and y1 and y2 are more preferably 1 to 6 (provided y1 + y2 = 2 to 11 is satisfied).

  The modified vinyl polymer according to this embodiment can be used as a blocking agent. The blocking agent according to this embodiment is suitable, for example, as a particle signal enhancer. Further, the blocking agent according to this embodiment may be the reaction product itself, or may contain a solvent as necessary.

  From the viewpoint of ease of handling when used in an aqueous solvent and the performance as a blocking agent described later, the modified vinyl polymer according to this embodiment is preferably water-dispersible or water-soluble, and more water-soluble. preferable. In the present invention, “water-soluble” means that it is transparent by visual observation when 1% of a polymer is dissolved in 20 ° C. water.

  More specifically, in the modified vinyl polymer according to this embodiment, oligoimine is introduced into the vinyl polymer, and the introduction site is preferably at both ends or one end of the vinyl polymer, and more preferably, at one end, oligoimine (after Will be described in detail).

  The oligoimine in the modified vinyl polymer usually contains a modified vinyl polymer in which one of the primary amino groups having two or more is bonded to the vinyl polymer, but a plurality of primary amino groups are bonded to the vinyl polymer. May be.

The modified vinyl polymer according to this embodiment is
(I) a method of reacting a terminal-reactive vinyl polymer obtained by living ion polymerization with oligoimine,
(Ii) a method in which a radical-initiating functional group such as an azo group is introduced into oligoimine and a vinyl polymer is radically polymerized using the functional group,
(Iii) a method of introducing a chain transfer functional group such as a mercapto group into an oligoimine and radically polymerizing a vinyl polymer using the functional group;
(Iv) a method in which a vinyl polymer obtained by radical polymerization of a vinyl monomer using a radical initiator having a functional group is reacted with oligoimine,
(V) a method of reacting an oligoimine with a vinyl polymer having a terminal functional group introduced by radical telomerization,
From the viewpoint of reaction time, molecular weight control, ease of production, etc., it is preferably produced by the method (v).

1.2. Production of Modified Vinyl Polymer (Blocking Agent) The modified vinyl polymer according to this embodiment is a vinyl polymerized in the presence of a compound having at least one group selected from the group consisting of a carboxyl group, an amino group and a hydroxyl group and a mercapto group. It can be obtained by reacting a polymer with an oligoimine.

  More specifically, the modified vinyl polymer according to this embodiment can be obtained by a reaction between a vinyl polymer having a terminal functional group introduced by radical telomerization and an oligoimine. The obtained modified vinyl polymer is preferably purified by a method such as dialysis, reprecipitation, or chromatography.

1.2.1. Oligoimine Oligoimine is a branched or straight-chain oligomer having an imino group and an alkylene group in the oligomer main chain and a primary amino group at two or more terminals, and has a molecular weight of 1000 or less (preferably a molecular weight of 500 or less). Specifically, dimethylenetrimine, trimethylenetetramine, tetramethylenepentamine, pentamethylenehexamine, hexamethyleneheptamine, heptamethyleneoctamine, octamethylenenonamine, nonamethylenedecamine, decamethyleneundecamine, unmethylene Oligomethyleneimines such as decamethylene dodecamine; diethylenetrimine, triethylenetetramine, tetraethylenepentamine, pentaethylenehexamine, hexaethyleneheptamine, heptaethyleneoctamine, octaethylenenonamine, Examples include oligoethyleneimines such as naethylene decamin, decaethylene undecamine, undecaethylene dodecamine, Epomin SP-003 (manufactured by Nippon Shokubai Co., Ltd.), and propylene-butylene-based imines such as spermine and spermidine. These can be used alone or in combination of two or more.

In view of solubility in water as a blocking agent and noise reduction effect, the oligoimine is preferably a linear oligoethyleneimine represented by the following general formula (4).
H ₂ N (CH ₂ CH ₂ NH) _n CH ₂ CH ₂ NH ₂ (4)
[In formula, n shows the number of 1-10. ]
In the said General formula (4), n becomes like this. Preferably it is an integer of 2-6, More preferably, it is 4.

1.2.2. Vinyl Polymer Introduced with Terminal Functional Group The vinyl polymer used for producing the modified vinyl polymer according to this embodiment preferably has a terminal functional group introduced by radical telomerization.

  Radical telomerization is a polymerization method in which a telogen, that is, a chain transfer agent having a functional group, is used during radical polymerization in order to introduce a specific functional group into a polymer terminal.

  Specific examples of the telogen include those that introduce a carboxyl group such as mercaptoacetic acid, 3-mercaptopropionic acid, 2-mercaptopropionic acid; 2-mercaptoethylamine, 3-mercaptopropylamine, 2-mercaptopropylamine, etc. What introduce | transduces an amino group; What introduce | transduces hydroxyl groups, such as 2-mercaptoethanol, 3-mercapto-1-propanol, 1-mercapto-2-propanol, 4-mercapto-1-butanol, is mentioned.

  Radical polymerization in radical telomerization is carried out by using the telogen and vinyl monomer and stirring and heating together with known solvents, initiators and the like. The polymerization time is usually 30 minutes to 24 hours, and the polymerization temperature is about 0 to 120 ° C.

  A vinyl polymer into which a terminal functional group is introduced (hereinafter referred to as “raw polymer”) is obtained by polymerizing a vinyl monomer having at least one unsaturated double bond in one molecule. The molecular weight is usually 1,000 to 1,000,000, preferably 2,000 to 100,000, and more preferably 3,000 to 50,000.

  Examples of the vinyl monomer that can be used for producing the raw material polymer include hydrophilic monomers and hydrophobic monomers, and hydrophilic monomers are preferable.

  Examples of hydrophilic monomers include acrylamide, N-hydroxyethylacrylamide, N, N-dimethylacrylamide, N-isopropylacrylamide, acryloylmorpholine, glycerol acrylate, glycerol methacrylate, polyethylene glycol (meth) acrylic ester, diacetone acrylamide, acrolein, Nonionic hydrophilic monomers such as 2-hydroxyethyl (meth) acrylate, N-vinylacetamide, N-vinyl-2-pyrrolidone; acrylic acid, methacrylic acid, itaconic acid, maleic acid, crotonic acid, styrenesulfonic acid, Anionic monomers such as isoprenesulfonic acid and 2-acrylamido-2-methylpropanesulfonic acid; allylamine, aminostyrene, N, N-dimethylamino B pills acrylamide, N, N-cationic monomers such as methyl chloride quaternary salt of dimethylaminopropyl acrylamide.

  Examples of hydrophobic monomers include methoxyethyl (meth) acrylate, methyl (meth) acrylate, ethyl (meth) acrylate, propyl (meth) acrylate, lauryl (meth) acrylate, cyclohexyl (meth) acrylate, ( Mention may be made of isobonyl (meth) acrylate, benzyl (meth) acrylate and styrene.

  The raw material polymer is preferably a homopolymer of these monomers and a copolymer obtained by polymerizing two or more kinds of monomers. The raw material polymer is preferably composed of 50% by weight or more of a nonionic hydrophilic monomer, more preferably 80% by weight or more of a nonionic hydrophilic monomer, and 95% by weight or more of nonionic. Most preferably, it is composed of a hydrophilic monomer. As the nonionic hydrophilic monomer, acrylamide and N-hydroxyethylacrylamide are particularly preferable, and acrylamide is most preferable. The raw material polymer can give a high signal by copolymerizing 1 to 10% by weight of an anionic monomer, particularly styrenesulfonic acid, isoprenesulfonic acid, 2-acrylamido-2-methylpropanesulfonic acid and the like.

1.2.3. Reaction of vinyl polymer introduced with terminal functional group by radical telomerization and oligoimine Reaction of vinyl polymer introduced with terminal functional group by radical telomerization and oligoimine includes, for example, vinyl carboxyl-terminated vinyl. A method in which a polymer is activated esterified with water-soluble carbodiimide or N-hydroxysuccinimide, and this is reacted with oligoimine; a vinyl polymer terminated with an amino group is converted into 1,3-butadiene diepoxide or 1,2,7,8-diepoxyoctane. And a method of reacting this with oligoimine; a method of tosylating a hydroxyl-terminated vinyl polymer with p-toluenesulfonic acid chloride and the like and reacting with oligoimine. The reaction is preferably a reaction in which a carboxyl group-terminated vinyl polymer is active esterified and bound to oligoimine.

1.3. Use of Modified Vinyl Polymer The modified vinyl polymer according to the present embodiment effectively reduces non-specific adsorption and reduces noise by substituting albumin, casein, gelatin, etc. used in conventional immunodiagnostic measurements. In addition, it can be used as a blocking agent that enhances the signal.

  For example, in the plate method, after binding a probe such as an antibody to the plate, the blocking agent according to this embodiment can be added to treat the plate surface.

  Moreover, the blocking agent which concerns on this embodiment can be used conveniently for manufacture of probe coupling | bonding particle | grains, for example.

1.4. Probe-bound particle and production thereof The method for producing a probe-bound particle according to the present embodiment processes the particle to which the probe is bound using the step of binding the probe to the surface of the particle and the blocking agent according to the present embodiment. Process.

  Here, the process of processing the particle | grains with which the probe was couple | bonded using the blocking agent which concerns on this embodiment can be performed by making the blocking agent which concerns on this embodiment contact the particle | grain surface for a predetermined time, for example. . Thereby, nonspecific adsorption | suction on the particle | grain surface can be suppressed and noise can be reduced. Moreover, this process can be performed in the state which disperse | distributed the particle | grains in the solution of the blocking agent which concerns on this embodiment, for example.

  In this case, the particle to which the probe is bound preferably has at least one group selected from an active ester group, a tosyl group and an epoxy group on the surface (the reason will be described later). In this case, the particles are preferably magnetic particles.

  More specifically, for example, in an immunochromatography method, after binding a probe such as an antibody to a colored particle, the blocking agent according to this embodiment is added to treat the surface of the colored particle, thereby binding the probe-bound particle. Can be obtained. Further, for example, in an assay method such as EIA, CLIA, CLEIA, etc., after binding a probe such as an antibody to the surface of the magnetic particle, the blocking agent according to this embodiment is added to treat the surface of the magnetic particle. Thus, probe-bound particles can be obtained.

  As described above, the blocking agent according to the present embodiment has a group represented by the general formula (1), thereby suppressing nonspecific adsorption, reducing noise, and false positive in immunodiagnostic measurement. The appearance rate of can be reduced.

  When the particles having at least the surface of active groups such as active ester groups, tosyl groups, and epoxy groups (for example, magnetic particles) are treated using the blocking agent according to this embodiment, the amino groups in the blocking agent and the particle surface Since the active group forms a covalent bond and improves the orientation of the antibody for immunodiagnosis, the effect of enhancing the signal appears. Further, since the elimination of the blocking agent can be suppressed, the test reagent is extremely stable with respect to buffers containing a surfactant or the like.

  The carrier in which the blocking agent according to the present embodiment exhibits a particularly good effect is particles having an active ester group (for example, magnetic particles). As a preferable treatment method, the particle having a carboxyl group is made into an active ester group in a water-soluble carbodiimide single system or a combination system of water-soluble carbodiimide and N-hydroxysuccinimide, and this is performed after binding an immunodiagnostic probe. The method of processing with the blocking agent (signal enhancer) which concerns on a form is mentioned.

  The probe-coupled particles according to this embodiment can be obtained by the method for producing probe-coupled particles. For example, the probe binding particles according to the present embodiment can have the blocking agent on the surface.

  The modified vinyl polymer of the present invention can be used for drug delivery system dispersing agents, coating agents for reducing non-specific adsorption to proteins and DNA, protein introduction and gene introduction in addition to the above blocking agents. It is also useful as a transfection reagent.

2. Examples Hereinafter, the present invention will be described in more detail with reference to examples, but the present invention is not limited thereto.

2.1. Example 1
900 g of water, 98 g of acrylamide, 2 g of 2-acrylamido-2-methylpropanesulfonic acid, and 2 g of 3-mercaptopropionic acid as a telogen were placed in a separable flask equipped with a stirrer, heated to 80 ° C. with stirring, and then used as an initiator. 1 g of 4,4′-azobiscyanovaleric acid was added and polymerization was continued for 3 hours with stirring. The obtained aqueous polymer solution was a colorless and transparent liquid, and as a result of measurement, the solid content was 10.5% (polymerization conversion rate 100%). This aqueous polymer solution was dialyzed with a dialysis tube having a molecular weight cut off of 3,500 for 48 hours while exchanging water, and then freeze-dried to obtain a vinyl polymer (A-1) as a raw material polymer.

  Next, 0.46 g of N-hydroxysuccinimide was put into a 100 cc container, 25 g of a 4% aqueous solution of vinyl polymer (A-1) was added and mixed therewith, and 1-ethyl-3-dimethylaminopropylcarbodiimide hydrochloride 0. 76 g was added and dissolved (active esterified polymer solution). In another container, 0.93 g of pentaethylenehexamine and 10 g of water were dissolved, and the active esterified polymer solution was added thereto, followed by stirring for 16 hours to carry out terminal modification. 10 g of water was added to the resulting solution, and this was dialyzed for 48 hours using a dialysis tube with a molecular weight cut off of 3,500 while exchanging water to obtain a modified vinyl polymer aqueous solution. Hereinafter, this is referred to as a blocking agent (B-1). The solid content of the obtained blocking agent (B-1) aqueous solution was 1.6%, and the number average molecular weight was 7800 and the weight average molecular weight was 14700 measured by GPC. The aqueous blocking agent (B-1) solution was freeze-dried, dissolved in heavy water at 5%, and subjected to NMR measurement using HDO = 4.24 ppm as a standard. As a result, four peaks (1. 85ppm, 1.70ppm, 1.29ppm, 1.17ppm) and peaks characteristic of pentaethylenehexamine (divided into about 7 peaks centering on the maximum peak of 2.93ppm). These peak area ratios and GPC From the obtained number average molecular weight, it was confirmed that 61% of the vinyl polymer (A-1) was reacted with pentaethylenehexamine.

  Next, an aqueous solution of 1-ethyl-3-dimethylaminopropylcarbodiimide hydrochloride was added to an aqueous dispersion having a solid content concentration of 1% in which 1 mg of magnetic particles having a carboxyl group (MAG2303 manufactured by JSR Corporation) were dispersed, and the mixture was stirred at room temperature. The carboxyl group was converted into an active ester by stirring for a period of time. This was magnetically separated and the supernatant was discarded, and then 10 μg of an antibody against human α-fetoprotein (hereinafter referred to as “AFP”), which is a tumor marker (hereinafter referred to as “anti-AFP antibody”, manufactured by Cosmo Bio). For 3 hours at room temperature. After the reaction, 125 μL of an aqueous dispersion of the above particles was added to a 0.4% aqueous solution of the blocking agent (B-1), and the mixture was further reacted at room temperature for 15 hours. This was magnetically separated, washed repeatedly with a cleaning solution (containing 25 mmol / L Tris-HCl, pH 7.4, 0.01% Tween 20), diluted with a cleaning solution to a particle concentration of 0.5%, and used as a primary probe. Probe-bound particles (immunoassay particles) bound with an anti-AFP antibody were obtained. Take 10 μl of the probe-bound particle dispersion (corresponding to 50 μg of particles) in a test tube and mix with 50 μl of a standard specimen of AFP antigen (manufactured by Nippon Biotest) diluted to 500 ng / mL with fetal calf serum (FCS). And allowed to react at 37 ° C. for 10 minutes. After separating the particles by magnetic separation and removing the supernatant, an anti-AFP antibody labeled with alkaline phosphatase (hereinafter referred to as “ALP”) as a secondary antibody (manufactured by Fujirebio Inc., attached to Lumipulse AFP-N) Using reagent) 40 μl was added and allowed to react at 37 ° C. for 10 minutes. Next, after magnetic separation and removal of the supernatant, the particles obtained by repeating washing three times with PBS were dispersed in 50 μl of 0.01% Tween 20, and transferred to a new tube. After adding 100 μl of ALP substrate solution (Lumipulse substrate solution: manufactured by Fujirebio Inc.) and reacting at 37 ° C. for 10 minutes, the amount of chemiluminescence was measured.

  For the measurement of chemiluminescence, a chemiluminescence measuring device (trade name: Lumat LB9507) manufactured by Bertol Japan KK was used. As a result, the signal intensity was 252497 RIU. The noise intensity was measured in the same manner as above except that 50 μl of FCS not containing AFP antigen was used instead of 50 μl of the standard specimen of AFP antigen diluted to 100 ng / mL with fetal calf serum (FCS). Met.

2.2. Example 2
In Example 1, a blocking agent (B-2) was obtained in the same manner as in Example 1 except that N-hydroxyethylacrylamide was used instead of acrylamide.

  Moreover, except having used the blocking agent (B-2) instead of the blocking agent (B-1), the magnetic particle was processed by the method similar to Example 1, and signal strength and noise strength were calculated | required. As a result, the signal intensity was 229661 RIU and the noise intensity was 256 RIU.

2.3. Example 3
In Example 1, in the same manner as in Example 1, except that a mixed monomer of 50 g of acryloylmorpholine and 50 g of polyethylene glycol methacrylate was used instead of 98 g of acrylamide and 2 g of 2-acrylamido-2-methylpropanesulfonic acid, A blocking agent (B-3) was obtained.

  Moreover, the magnetic particle was processed by the method similar to Example 1 except having used the blocking agent (B-3) instead of the blocking agent (B-1), and signal strength and noise strength were calculated | required. As a result, the signal intensity was 215974 RIU, and the noise intensity was 249 RIU.

2.4. Comparative Example 1
The blocking agent (B-4) of Comparative Example 1 was obtained in the same manner as in Example 1 except that ethylenediamine was used instead of pentaethylenehexamine in Example 1.

  Moreover, the magnetic particle was processed by the method similar to Example 1 except having used the blocking agent (B-4) instead of the blocking agent (B-1), and signal strength and noise strength were calculated | required. As a result, the signal intensity was 204656RIU and the noise intensity was 252RIU.

2.5. Comparative Example 2
In Example 1, except that bovine serum albumin was used instead of the blocking agent (B-1), the magnetic particles were processed in the same manner as in Example 1 to determine the signal intensity and noise intensity. As a result, the signal intensity was 214082 RIU and the noise intensity was 315 RIU.

2.6. Comparative Example 3
In Example 1, except that the blocking agent (B-1) was not used, the magnetic particles were processed in the same manner as in Example 1 to determine the signal intensity and noise intensity. The signal intensity was 193569 RIU and the noise intensity was 321 RIU.

Claims (9)

A water-soluble vinyl polymer having a group represented by the following general formula (1), which is polymerized in the presence of a compound having at least one group selected from the group consisting of a carboxyl group, an amino group and a hydroxyl group and a mercapto group Water-soluble vinyl polymer obtained by the reaction of the prepared vinyl polymer with oligoimine .

(1)
[Wherein, R ¹ and R ² independently represent a hydrogen atom, an amino group and an imino group, or a group having 1 to 11 of either one (provided that the amino group contained in R ¹ and R ² and The total number of imino groups is 2 to 11.)] The water-soluble vinyl polymer according to claim 1, wherein, in the general formula (1), R ¹ and R ² are groups represented by the following general formulas (2a) and (2b), respectively.

(2a)

(2b)
[Wherein, x1 and x2 are each a number represented by 1 to 4, and y1 and y2 are each a number represented by 0 to 11 (provided that y1 + y2 satisfies 2 to 11)] The water-soluble vinyl polymer according to claim 1 or 2 , wherein the oligoimine is a compound represented by the following general formula (4).
H ₂ N (CH ₂ CH ₂ NH) _n CH ₂ CH ₂ NH ₂ (4)
[In formula, n shows the number of 1-10. ] A blocking agent comprising a water-soluble vinyl polymer having a group represented by the following general formula (1) .

(1)
[Wherein, R ¹ and R ² independently represent a hydrogen atom, an amino group and an imino group, or a group having 1 to 11 of either one (provided that the amino group contained in R ¹ and R ² and The total number of imino groups is 2 to 11.)] Binding a probe to the surface of the particle;
Treating the particles to which the probe is bound using the blocking agent of claim 4 ;
A method for producing probe-bound particles, comprising: The method for producing probe-bonded particles according to claim 5 , wherein, in the step of bonding the probes, the particles have at least one group selected from an active ester group, a tosyl group, and an epoxy group. The method for producing probe-bonded particles according to claim 5 , wherein the particles are magnetic particles. Probe-bound particles obtained by the method for producing probe-bound particles according to claim 5 . Probe binding particles having the blocking agent according to claim 4 on the surface.