CN1152053C - Method for induction and prepn. of epitope idiosyncratic monoclonal antibody by using epitope polypeptide - Google Patents
Method for induction and prepn. of epitope idiosyncratic monoclonal antibody by using epitope polypeptide Download PDFInfo
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- CN1152053C CN1152053C CNB991238087A CN99123808A CN1152053C CN 1152053 C CN1152053 C CN 1152053C CN B991238087 A CNB991238087 A CN B991238087A CN 99123808 A CN99123808 A CN 99123808A CN 1152053 C CN1152053 C CN 1152053C
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- epitope
- polypeptide
- epitope polypeptide
- protein
- monoclonal antibody
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Abstract
The present invention belongs to the technical field of biological engineering. Firstly, epitope polypeptide with particular epitopes on nature protein or gene recombination protein is artificially synthesized, one or a plurality of epitopes repeatedly appear on the polypeptide artificially synthesized (namely the epitope polypeptide), and the epitope polypeptide is coupled on carrier protein or carrier polypeptide; the coupled substances of the epitope polypeptide are matched with proper adjuvant to immunize animals, and hybridoma is prepared by a conventional cell fusion technology. Under the condition of no native protein or gene recombination protein, the present invention can directly induce/prepare monoclonal antibodies with predetermined epitope specificity in high efficiency.
Description
Technical field the invention belongs to technical field of bioengineering, particularly the Monoclonal Antibody of epitope specificity and production.
Background technology monoclonal antibody technique (hybridoma technology) is invention in 1975, the acquisition Nobel prize in 1984.This technology has produced epoch-making influence to life science and medical research, makes the diagnosis of disease and treatment technology produce revolutionary variation, and has brought up global annual 10000000000 dollars industry.Current at present monoclonal antibody technique is the protein immune animal that utilizes native protein or gene recombination, prepares the hybridoma clone of secrete monoclonal antibody then by cell-fusion techniques.
Existing monoclonal antibody technique has two difficult problems that can't overcome: one, the albumen of native protein or gene recombination can't obtain or be difficult to extracting, thereby can not prepare monoclonal antibody; Its two, the monoclonal antibody that will just can obtain separating the epitope specificity that obtains needs through a large amount of screenings often.
Summary of the invention the objective of the invention is for solving the difficult problem that prior art exists, the monoclonal antibody method of epitope specificity is induced/is prepared in proposition with epitope polypeptide, can not have directly to induce/prepare predetermined epitope specificity monoclonal antibody efficiently under the proteic condition of native protein or gene recombination.
What the present invention proposed induces/prepares the monoclonal antibody method of epitope specificity with epitope polypeptide, it is characterized in that may further comprise the steps:
1) synthetic has the repetition epitope polypeptide C-(GPGRAFY) on native protein or the gene recombinant protein
4
2) said epi-position often is to appear on the polypeptide (being epitope polypeptide) of synthetic with the multiple form;
3) said epitope polypeptide is coupled on carrier proteins or the carrier polypeptide;
4) coupling matter of said epitope polypeptide is equipped with and is fit to the adjuvant immunity animal;
5) adopt conventional cell-fusion techniques to prepare hybridoma.
The effect of invention: we have used this technology successfully to prepare the monoclonal antibody of two strain epitope specificities.
Embodiment
Embodiment: the specific monoclonal antibody of preparation GPGRAFY
Step 1, the synthetic epitope polypeptide C-(GPGRAFY) that repeats
4
Step 2 utilizes MBS (m-maleimidoberrzoyl-N-hydroxy succinimide ester) with epitope polypeptide and carrier proteins BSA coupling connection;
Step 3 joins the epitope polypeptide and the freund adjuvant mixed immunity Balb/c female mice of carrier with coupling, uses complete freund adjuvant for the first time, and full freund adjuvant toos many or too much for use later on.Immunizing dose be 10 μ g/ each/every; Per two all immunity once;
Step 4, the cytogamy experiment;
Step 5, the hybridoma screening.
Claims (1)
1, a kind ofly induce/prepare the monoclonal antibody method of epitope specificity, it is characterized in that may further comprise the steps with epitope polypeptide:
1) synthetic has the repetition epitope polypeptide C-(GPGRAFY) on native protein or the gene recombinant protein
4
2) said epi-position is to appear on the epitope polypeptide of synthetic with the multiple form;
3) said epitope polypeptide is coupled on carrier proteins or the carrier polypeptide;
4) coupling matter of said epitope polypeptide is equipped with suitable adjuvant immunity animal;
5) adopt conventional cell-fusion techniques to prepare hybridoma.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB991238087A CN1152053C (en) | 1999-11-12 | 1999-11-12 | Method for induction and prepn. of epitope idiosyncratic monoclonal antibody by using epitope polypeptide |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB991238087A CN1152053C (en) | 1999-11-12 | 1999-11-12 | Method for induction and prepn. of epitope idiosyncratic monoclonal antibody by using epitope polypeptide |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN1259524A CN1259524A (en) | 2000-07-12 |
| CN1152053C true CN1152053C (en) | 2004-06-02 |
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Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNB991238087A Expired - Fee Related CN1152053C (en) | 1999-11-12 | 1999-11-12 | Method for induction and prepn. of epitope idiosyncratic monoclonal antibody by using epitope polypeptide |
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| CN (1) | CN1152053C (en) |
Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110790829B (en) * | 2017-12-19 | 2022-08-02 | 北京泽勤生物医药有限公司 | Application of antibody prepared by using pHLIP extracellular segment as antigen in preparation of antitumor drugs |
| CN110272502B (en) * | 2019-07-12 | 2021-09-14 | 深圳市亚辉龙生物科技股份有限公司 | Immunogen, hybridoma cell secreting anti-cardiac troponin I monoclonal antibody, preparation method, monoclonal antibody and application |
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1999
- 1999-11-12 CN CNB991238087A patent/CN1152053C/en not_active Expired - Fee Related
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| Publication number | Publication date |
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| CN1259524A (en) | 2000-07-12 |
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