CN107893051A - A kind of method of excretion body in serum using immuno magnetic cell separation - Google Patents
A kind of method of excretion body in serum using immuno magnetic cell separation Download PDFInfo
- Publication number
- CN107893051A CN107893051A CN201710940765.7A CN201710940765A CN107893051A CN 107893051 A CN107893051 A CN 107893051A CN 201710940765 A CN201710940765 A CN 201710940765A CN 107893051 A CN107893051 A CN 107893051A
- Authority
- CN
- China
- Prior art keywords
- magnetic
- volume
- serum
- incubated
- suspension
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
- C12N5/06—Animal cells or tissues; Human cells or tissues
- C12N5/0602—Vertebrate cells
- C12N5/0634—Cells from the blood or the immune system
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2509/00—Methods for the dissociation of cells, e.g. specific use of enzymes
Landscapes
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Biomedical Technology (AREA)
- Life Sciences & Earth Sciences (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Wood Science & Technology (AREA)
- Biotechnology (AREA)
- Organic Chemistry (AREA)
- Chemical & Material Sciences (AREA)
- Genetics & Genomics (AREA)
- Zoology (AREA)
- Hematology (AREA)
- Microbiology (AREA)
- Immunology (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Cell Biology (AREA)
- Peptides Or Proteins (AREA)
Abstract
The invention provides a kind of method of excretion body in serum using immuno magnetic cell separation, this method comprises the following steps：1) serum pre-processes；2) immunomagnetic beads is prepared；3) by serum pretreatment product obtained by step 1) and 2), the middle immunomagnetic beads obtained is incubated, and obtains antibody bead complexes absorption excretion body.Advantages of the present invention：It is simple to operate, resulting product purity it is high, it is necessary to equipment it is relatively simple, and the excretion body containing special sign thing in serum can be separated.
Description
Technical field
The present invention relates to biomedical sector, in particular it relates to excretion body in a kind of serum using immuno magnetic cell separation
Method.
Background technology
Excretion body (exosome) is a kind of small film bubble that can be secreted by most cells, has double-layer of lipoid membrane structure,
Diameter about 40-100nm, plays and acts in biological processes and intercellular signal transmission.Contain on excretion body film surface
The marker proteins such as CD9, CD63 and CD81, film is interior to contain the large biological molecules such as nucleic acid such as microRNA.At present, serum excretion body point
From mainly having based on ultracentrifugal isolation technics, isolation technics (chromatography), the PEG precipitation method based on molecular size, it is based on
Isolation technics of immuno absorbence etc..The excretion body yield that centrifugal process obtains is higher, but it takes time and effort, and is not suitable for great amount of samples behaviour
Make.The excretion body purity of chromatography extraction is higher, but its separation is easily disturbed by foreign protein, and operational stability is poor, and generally to sample
This can produce diluting effect, and obtained excretion bulk concentration is relatively low, it is difficult to carries out subsequent detection.The excretion body that the PEG precipitation method obtain
It is time-consuming short, but gained excretion body purity is relatively low.Importantly, above-mentioned centrifugal process, chromatography and the PEG precipitation method isolate and purify blood
Total excretion body in clear, and can not separation and Extraction contain the excretion body (such as CD9+/CD63+ excretions body) of special sign thing.
Substantial amounts of protein and acceptor on excretion body film be present, the affine in immunity between these protein (antigen-antibody) is made
With and idiosyncrasy between acceptor and part, provide new method for effectively special separation excretion body.Immunomagnetic beads point
From technology with its targeting specific it is strong, it is easy to operate, separation it is efficient the advantages of, be widely used in biochemistry, molecule, science of heredity,
The every field such as pathology, physiology, pharmacology, microorganism.At present, U.S. SBI companies, Japanese MBL companies have been developed based on magnetic bead
Affine in immunity capture technique is used for isolating and purifying for excretion body, and the excretion that special sign thing is expressed in serum is separated available for extraction
Body.
The content of the invention
It is an object of the present invention to provide a kind of method of excretion body in serum using immuno magnetic cell separation, this method includes following
Step：
1) pre-process, (room temperature) takes fresh serum, separated through ultracentrifugation, tentatively go the removal of impurity, take supernatant；
Further, step 1) includes：Fresh serum is taken, room temperature 3000g is centrifuged 30 minutes, abandons pellet cell debris；Take
Centrifuged supernatant, the PBS for adding 29 times of volumes are diluted, and 4 DEG C of 100000g, are centrifuged 2 hours, are collected precipitation, are resuspended.
2) suspension containing magnetic beads are taken to be cleaned with appropriate dissociating buffer, magnetic frame separation magnetic bead, with the appropriate dissociating buffer
Suspend；Then appropriate CD5L monoclonal antibodies are added into gained suspension to be incubated；With appropriate (such as 100 times after the completion of incubation
Volume) dissociating buffer cleaning, magnetic frame separation magnetic bead (at least cleaning three times), then with right amount (such as 100 times of volumes)
Dissociating buffer suspends；
Further, using the magnetic bead of Streptavidin modification, such as Streptavidin modification in step 2)Magnetic bead.
Further, the dosage of dissociating buffer is 50-500 times of suspension containing magnetic beads volume in step 2), such as 100 times.
Further, the addition volume of CD5L monoclonal antibodies described in step 2) and the volume ratio of the suspension containing magnetic beads are
1-1.5:50-100。
Further, incubation time described in step 2) at least 12 hours.
3) 2-10 times of debulking step 1 is added in the bead suspension finally given to step 2)) obtained supernatant is pre-processed,
It is incubated；
Further, incubation time described in step 3) at least 24 hours.
4) system after the completion of step 3) is incubated is cleaned with appropriate (such as 100 times of volumes) phosphate buffered saline solution, magnetic
Power frame separates magnetic bead, that is, obtains being adsorbed with the compound of excretion body, i.e. magnetic bead-antibody-excretion nanocrystal composition.
Further, the dissociating buffer is the phosphate buffered saline solution containing 0.1% bovine serum albumin(BSA).
Further, the dissociating buffer is in advance with 0.22 micron membrane filter filtration sterilization.
Such as indicated without special, incubation conditions of the present invention are：37 DEG C of temperature, couveuse parameter are rotated by 90 ° to be reverse, inclined
Oblique 5 seconds, 5 ° of vibrations continued 1 second.
Specifically, in the serum of the present invention using immuno magnetic cell separation excretion body method, including following steps：
1) pre-process, at room temperature with fresh serum, separated through ultracentrifugation, tentatively go the removal of impurity, take supernatant；
2) 1 times of volume suspension containing magnetic beads is taken to separate buffer solution for cleaning, magnetic frame separation magnetic bead, 100 times of volumes with 100 times of volumes
Dissociating buffer suspends；Above-mentioned suspension is added into 1/10 times of volume CD5L monoclonal antibody to be incubated 12 hours；After the completion of incubation
Buffer solution for cleaning is separated with 100 times of volumes, magnetic frame separation magnetic bead, is cleaned three times, 100 times of volume dissociating buffers suspend；
3) 2-10 times of debulking step 1 is added in the bead suspension for finally giving step 2)) handle obtained pretreatment
Supernatant, it is incubated 24 hours；4) cleaned after the completion of step 3) is incubated with 100 times of volume phosphate buffered saline solutions, magnetic frame separation
Magnetic bead, that is, the excretion nanocrystal composition contained (i.e. magnetic bead-antibody-excretion nanocrystal composition).
The technical characterstic of the present invention is：
The basic thought of the present invention is to use excretion body in immuno magnetic cell separation serum.
Excretion body in serum, is enriched with by the defects of present invention is directed to prior art with the method for immunomagnetic beads, is operated
Simply, resulting product purity is high, it is not necessary to particular device, can pointedly separate the excretion of expression specified protein
Body.
Brief description of the drawings
Fig. 1 is the Dot-blot of CD5L+ excretion bodies in immuno magnetic cell separation serum.
Fig. 2 is the Western-blot of CD5L+ excretion bodies in immuno magnetic cell separation serum.
Embodiment
Following examples are used to illustrate the present invention, but are not limited to the scope of the present invention.It is unreceipted specific in embodiment
Technology or condition person, carried out according to the technology or condition described by document in the art, or according to product description.It is used
Reagent or the unreceipted production firm person of instrument, it is the conventional products that can be commercially available by regular distributor.
The key instrument and reagent used in following embodiments：M-280Tosylactivated
(Ancell companies, the U.S.)；Rabbit-anti people CD5L polyclonal antibodies (Abcam companies, the U.S.)；HRP mark secondary antibodies (Jackson
Immuno Research)；Ultracentrifuge (Beckman coulter, the U.S.)；Magnetic frame (NewEnglandBiolabs)；
Half-dried transferring film instrument, electrophoresis tank are purchased from Bio-rad.
CD5L+ excretion bodies in the immuno magnetic cell separation serum of embodiment 1
1st, experimental procedure：
1) fresh serum is collected, is placed in a centrifuge, room temperature 3000g is centrifuged 30 minutes, is abandoned pellet cell debris, is taken centrifugation
Supernatant, with 1:29 (serum:PBS volume ratio) is diluted, using ultracentrifuge carry out ultracentrifugation (100000g,
2 hours, 4 DEG C), precipitation is collected, is resuspended.
2) whirlpool concussion Streptavidin modificationMagnetic bead, after bead suspension is homogeneous, take 165 micro-
Rise suspension containing magnetic beads to be added in 1ml dissociating buffers, suspend cleaning, magnetic frame separation magnetic bead, repeated washing three times.Cleaning is completed
Afterwards, 250 liters of dissociating buffer suspension magnetic beads.
Take 6 microlitres of rabbit-anti people's CD5L polyclonal antibodies to be added in the suspension containing magnetic beads of above-mentioned gained, overturn and be well mixed, put
In being incubated on couveuse 12 hours (parameter is rotated by 90 ° to be reverse, 37 DEG C)；It is incubated and completes, Incubating Solution is placed on magnetic frame 2 points
Clock, clear liquid is abandoned, magnetic bead is suspended with 1ml dissociating buffers, and suspend cleaning, magnetic frame separation magnetic bead, is cleaned three times.Cleaning is completed
Afterwards, 250 microlitres of dissociating buffer suspension magnetic beads.
3) take the precipitation re-suspension liquid obtained by 150 microgram steps 1) to be added in the suspension containing magnetic beads obtained by step 2), overturn mixed
Close uniformly, put and be incubated on couveuse 24 hours (parameter is rotated by 90 ° to be reverse, 4 DEG C)；
4) it is incubated and completes, Incubating Solution is placed on magnetic frame, abandons clear liquid, and magnetic bead is suspended with 1ml dissociating buffers, is suspended clear
Wash, magnetic frame separation magnetic bead, clean three times；After the completion of cleaning, that is, obtain being adsorbed with the compound i.e. magnetic beads of CD5L+ excretion bodies-
Antibody-CD5L+ excretion nanocrystal compositions.
2nd, experimental result：
As Fig. 1 shows, be according to the above method it is separated obtain the compound i.e. magnetic bead-antibody for having a CD5L+ excretion bodies-
CD5L+ excretion nanocrystal compositions Dot-blot resulting after elution.It is highly enriched that shown result shows that excretion body obtains.
Exo is excretion body.
As Fig. 2 shows, be according to the above method it is separated obtain the compound i.e. magnetic bead-antibody for having a CD5L+ excretion bodies-
CD5L+ excretion nanocrystal compositions Western-blot resulting after elution.It is rich that shown result shows that excretion body obtains height
Collection.TSG101 is excretion body specificity marker, frequently as the specific proteins of excretion body identification.
Claims (9)
1. a kind of method of excretion body in serum using immuno magnetic cell separation, it is characterised in that comprise the following steps：
1) pre-process：Fresh serum is taken, is separated through ultracentrifugation, tentatively goes the removal of impurity, takes supernatant；
2) take suspension containing magnetic beads to be cleaned with appropriate dissociating buffer, magnetic frame separation magnetic bead, suspended with the appropriate dissociating buffer；
Then appropriate CD5L monoclonal antibodies are added into gained suspension to be incubated；It is clear with the appropriate dissociating buffer after the completion of incubation
Wash, magnetic frame separation magnetic bead, then suspended with appropriate dissociating buffer；
3) 2-10 times of debulking step 1 is added in the bead suspension finally given to step 2)) obtained supernatant is pre-processed, incubate
Educate；
4) system after the completion of step 3) is incubated is cleaned with appropriate phosphate buffered saline solution, magnetic frame separation magnetic bead, that is, is obtained
It is adsorbed with the compound of excretion body.
2. according to the method for claim 1, it is characterised in that the dissociating buffer is containing 0.1% bovine serum albumin(BSA)
Phosphate buffered saline solution；Preferably, the dissociating buffer is in advance with 0.22 micron membrane filter filtration sterilization.
3. method according to claim 1 or 2, it is characterised in that step 1) includes：Take fresh serum, room temperature 3000g from
The heart 30 minutes, abandons pellet cell debris；Centrifuged supernatant is taken, the PBS for adding 29 times of volumes is diluted, 4 DEG C of 100000g, from
The heart 2 hours, precipitation is collected, be resuspended.
4. according to the method described in claim any one of 1-3, it is characterised in that step 2) uses the magnetic of Streptavidin modification
Pearl；It is preferred that Streptavidin is modifiedMagnetic bead.
5. according to the method described in claim any one of 1-3, it is characterised in that the dosage of dissociating buffer is magnetic in step 2)
50-500 times of pearl suspension volume.
6. according to the method described in claim any one of 1-3, it is characterised in that CD5L's monoclonal antibodies described in step 2)
It is 1-1.5 to add volume and the volume ratio of the suspension containing magnetic beads:50-100.
7. according to the method described in claim any one of 1-3, it is characterised in that incubation time described in step 2) at least 12 is small
When；And/or
Incubation time described in step 3) at least 24 hours.
8. according to the method described in claim any one of 1-7, it is characterised in that the incubation conditions are：37 DEG C of temperature, it is incubated
Device parameter is rotated by 90 ° to be reverse, tilts 5 seconds, 5 ° of vibrations continue 1 second.
9. according to the method described in claim any one of 1-8, it is characterised in that including following steps：
1) pre-process, at room temperature with fresh serum, separated through ultracentrifugation, tentatively go the removal of impurity, take supernatant；
2) 1 times of volume suspension containing magnetic beads is taken to separate buffer solution for cleaning, magnetic frame separation magnetic bead, 100 times of volume separation with 100 times of volumes
Buffer solution suspends；Above-mentioned suspension is added into 1/10 times of volume CD5L monoclonal antibody to be incubated 12 hours；With 100 after the completion of incubation
Times volume separation buffer solution for cleaning, magnetic frame separation magnetic bead, clean three times, 100 times of volume dissociating buffers suspend；
3) 2-10 times of debulking step 1 is added in the bead suspension for finally giving step 2)) obtained pretreatment supernatant is handled,
It is incubated 24 hours；
4) cleaned, magnetic frame separation magnetic bead, that is, contained with 100 times of volume phosphate buffered saline solutions after the completion of step 3) is incubated
Some excretion nanocrystal compositions.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710940765.7A CN107893051A (en) | 2017-10-11 | 2017-10-11 | A kind of method of excretion body in serum using immuno magnetic cell separation |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710940765.7A CN107893051A (en) | 2017-10-11 | 2017-10-11 | A kind of method of excretion body in serum using immuno magnetic cell separation |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN107893051A true CN107893051A (en) | 2018-04-10 |
Family
ID=61802559
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201710940765.7A Pending CN107893051A (en) | 2017-10-11 | 2017-10-11 | A kind of method of excretion body in serum using immuno magnetic cell separation |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN107893051A (en) |
Cited By (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109116010A (en) * | 2018-08-22 | 2019-01-01 | 威海纽兰生物科技有限公司 | Test tube and excretion body separation method for the acquisition of blood excretion body |
| CN109266599A (en) * | 2018-10-16 | 2019-01-25 | 郑州大学 | A kind of excretion body separation method that efficient pattern is lossless |
| CN110079457A (en) * | 2019-06-04 | 2019-08-02 | 苏州大学 | Micro-fluidic chip and excretion body extracting method |
| CN110152747A (en) * | 2019-05-10 | 2019-08-23 | 清华大学 | The separation method of micro-fluidic chip and excretion body |
| CN110747158A (en) * | 2019-11-14 | 2020-02-04 | 赵凯 | Cell supernatant exosome extraction process based on precipitation reagent method |
| CN111505264A (en) * | 2019-01-30 | 2020-08-07 | 上海思路迪生物医学科技有限公司 | Exosome separation method, immunomagnetic beads and kit |
| CN111621416A (en) * | 2020-05-14 | 2020-09-04 | 南京大学 | Biological magnetic separation method based on mobile magnetic net |
| WO2021147127A1 (en) * | 2020-01-21 | 2021-07-29 | 武汉生之源生物科技股份有限公司 | Kit for separating exosomes from cell supernatant and use method of kit |
| CN113728233A (en) * | 2019-01-31 | 2021-11-30 | 积水医疗株式会社 | Immunoassay method and assay kit for free AIM in biological samples |
| CN115322954A (en) * | 2022-08-30 | 2022-11-11 | 柳州市人民医院 | Method for separating and purifying influenza virus infected epithelial cell exosomes |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101490085A (en) * | 2006-06-12 | 2009-07-22 | 特鲁比昂药品公司 | Single-chain multivalent binding proteins with effector function |
| CN106289926A (en) * | 2016-07-26 | 2017-01-04 | 华东理工大学 | A kind of method using immuno magnetic cell separation serum China and foreign countries to secrete body |
-
2017
- 2017-10-11 CN CN201710940765.7A patent/CN107893051A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101490085A (en) * | 2006-06-12 | 2009-07-22 | 特鲁比昂药品公司 | Single-chain multivalent binding proteins with effector function |
| CN106289926A (en) * | 2016-07-26 | 2017-01-04 | 华东理工大学 | A kind of method using immuno magnetic cell separation serum China and foreign countries to secrete body |
Non-Patent Citations (1)
| Title |
|---|
| ARMANDO DE MENEZES-NETO等: ""Size-exclusion chromatography as a stand-alone methodology identifies novel markers in mass spectrometry analyses of plasma-derived vesicles from healthy individuals"", 《JOURNAL OF EXTRACELLULAR VESICLES》 * |
Cited By (13)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109116010B (en) * | 2018-08-22 | 2023-11-21 | 威海纽兰生物科技有限公司 | Test tube for blood exosome collection and exosome separation method |
| CN109116010A (en) * | 2018-08-22 | 2019-01-01 | 威海纽兰生物科技有限公司 | Test tube and excretion body separation method for the acquisition of blood excretion body |
| CN109266599A (en) * | 2018-10-16 | 2019-01-25 | 郑州大学 | A kind of excretion body separation method that efficient pattern is lossless |
| CN111505264A (en) * | 2019-01-30 | 2020-08-07 | 上海思路迪生物医学科技有限公司 | Exosome separation method, immunomagnetic beads and kit |
| CN111505264B (en) * | 2019-01-30 | 2024-05-17 | 上海思路迪生物医学科技有限公司 | Exosome separation method, immunomagnetic beads and kit |
| CN113728233A (en) * | 2019-01-31 | 2021-11-30 | 积水医疗株式会社 | Immunoassay method and assay kit for free AIM in biological samples |
| CN110152747B (en) * | 2019-05-10 | 2020-06-02 | 清华大学 | Microfluidic chip and exosome separation method |
| CN110152747A (en) * | 2019-05-10 | 2019-08-23 | 清华大学 | The separation method of micro-fluidic chip and excretion body |
| CN110079457A (en) * | 2019-06-04 | 2019-08-02 | 苏州大学 | Micro-fluidic chip and excretion body extracting method |
| CN110747158A (en) * | 2019-11-14 | 2020-02-04 | 赵凯 | Cell supernatant exosome extraction process based on precipitation reagent method |
| WO2021147127A1 (en) * | 2020-01-21 | 2021-07-29 | 武汉生之源生物科技股份有限公司 | Kit for separating exosomes from cell supernatant and use method of kit |
| CN111621416A (en) * | 2020-05-14 | 2020-09-04 | 南京大学 | Biological magnetic separation method based on mobile magnetic net |
| CN115322954A (en) * | 2022-08-30 | 2022-11-11 | 柳州市人民医院 | Method for separating and purifying influenza virus infected epithelial cell exosomes |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN107893051A (en) | A kind of method of excretion body in serum using immuno magnetic cell separation | |
| CN106289926A (en) | A kind of method using immuno magnetic cell separation serum China and foreign countries to secrete body | |
| CN110231207B (en) | Method for separating exosome | |
| US10669535B2 (en) | Methods for the isolation of extracellular vesicles and other bioparticles from urine and other biofluids | |
| CN109891236A (en) | The method for recycling extracellular vesica | |
| CN110343664B (en) | Method for extracting exosome and exosome protein | |
| CN113101737B (en) | Affinity tangential flow filtration system, construction method thereof, exosome extraction method and application | |
| CN111321108A (en) | High-purity exosome density gradient centrifugation method | |
| CN109913417A (en) | A method of using different cell origin excretion body hypotypes in tachysynthesis paramagnetic particle method separation cerebrospinal fluid | |
| CN113249302A (en) | Efficient exosome separation and purification method | |
| Yoshida et al. | TIM4-affinity methods targeting phosphatidylserine for isolation or detection of extracellular vesicles | |
| CN101644704B (en) | Preparation method for rheumatoid factor detection reagent | |
| Childs III et al. | Use of Percoll density gradients for studying the attachment of bacteria to oral epithelial cells | |
| CN101125886A (en) | Method for extracting yolk immunoglobulin | |
| CN114276992A (en) | Complete exosome separation and purification kit and detection analysis method | |
| CN114574437B (en) | Plasma exosome extraction reagent, enrichment method, extraction kit and application thereof | |
| KR930012113B1 (en) | Method for purification of hbc antigen and method for measurement of hbc antibody | |
| CN115932250B (en) | Magnetic particle chemiluminescence reagent for quantitative detection of anti-aPS/PT antibody and chemiluminescence detection kit | |
| CN111394307A (en) | Method for separating and purifying exosome from plasma and application | |
| US20130288227A1 (en) | Methods and compositions for cell separation of blood tissues | |
| JP2008189648A (en) | New antigen derived from helicobacter pylori, antigenic composition, and detection method for helicobacter pylori antibody | |
| JPS6231301B2 (en) | ||
| CN117092354B (en) | Protein marker for identifying extracellular vesicles of brain-derived cells | |
| Viljoen et al. | Isolation of Cowdria ruminantium by cellular affinity chromatography and detection by an enzyme-linked immunosorbent assay | |
| Litman et al. | Rapid purification of IgA from normal human serum |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20180410 |