CN1259524A - 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
- Publication number
- CN1259524A CN1259524A CN 99123808 CN99123808A CN1259524A CN 1259524 A CN1259524 A CN 1259524A CN 99123808 CN99123808 CN 99123808 CN 99123808 A CN99123808 A CN 99123808A CN 1259524 A CN1259524 A CN 1259524A
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- China
- Prior art keywords
- polypeptide
- epi
- epitope
- protein
- monoclonal antibody
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Links
- 229920001184 polypeptide Polymers 0.000 title claims abstract description 25
- 108090000765 processed proteins & peptides Proteins 0.000 title claims abstract description 25
- 102000004196 processed proteins & peptides Human genes 0.000 title claims abstract description 25
- 238000000034 method Methods 0.000 title claims abstract description 13
- 230000006698 induction Effects 0.000 title 1
- 108090000623 proteins and genes Proteins 0.000 claims abstract description 15
- 102000004169 proteins and genes Human genes 0.000 claims abstract description 10
- 241001465754 Metazoa Species 0.000 claims abstract description 4
- 230000007910 cell fusion Effects 0.000 claims abstract description 4
- 102000014914 Carrier Proteins Human genes 0.000 claims abstract description 3
- 108010078791 Carrier Proteins Proteins 0.000 claims abstract description 3
- 239000002671 adjuvant Substances 0.000 claims description 5
- 210000004408 hybridoma Anatomy 0.000 claims description 5
- 230000008878 coupling Effects 0.000 claims description 4
- 238000010168 coupling process Methods 0.000 claims description 4
- 238000005859 coupling reaction Methods 0.000 claims description 4
- 230000036039 immunity Effects 0.000 claims description 4
- 108010008281 Recombinant Fusion Proteins Proteins 0.000 claims description 2
- 238000005215 recombination Methods 0.000 abstract description 5
- 230000006798 recombination Effects 0.000 abstract description 5
- 206010028980 Neoplasm Diseases 0.000 abstract 1
- 239000000126 substance Substances 0.000 abstract 1
- 238000005516 engineering process Methods 0.000 description 4
- 238000012216 screening Methods 0.000 description 2
- 241000699670 Mus sp. Species 0.000 description 1
- 230000024835 cytogamy Effects 0.000 description 1
- 238000003745 diagnosis Methods 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 150000002148 esters Chemical class 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 230000003053 immunization Effects 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
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- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Peptides Or Proteins (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
In the present invention we synthesize artificially epi-position polyeptide carrying natural protein or gene recombination protein with specific epi-position, its one or sereral epi-positions are aften repeatedly appeared on artificially-synthesized polypeptide (that is epi-rosition poly peptide): the epi-position polypeptide is coupled on to carrier protein or carrier polypeptide, the coupled substance of epi-position polypeptide with suitable adjurant immune animal, and the conventional cell fusion technique is adopted to prepare hybridized tumour. The present method can directly induce/prepare predetermined epi-position spsecific mono clone antibody effectively under the condition of having no natural protein or gene recombination protein.
Description
The invention belongs to technical field of bioengineering, particularly the Monoclonal Antibody of epitope specificity and production.
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.
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 epitope polypeptide of the defined epitope on native protein or the gene recombinant protein;
2) said one or more epi-position often be with the multiple form appear at synthetic polypeptide (promptly the table
The position polypeptide) on;
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: the specific monoclonal antibody of preparation GPGRAFY
Step 1, the synthetic epitope polypeptide C-(GPGRAFY) that repeats
4
Step 2, utilize MBS (mmaleimidobenzoyl-N-hydroxy suocinimide ester) with epitope polypeptide with carry
Body protein BSA coupling connection;
Step 3 joins the epitope polypeptide and the freund adjuvant mixed immunity Balb/c female mice of carrier with coupling,
Once use complete freund adjuvant, full freund adjuvant toos many or too much for use later on.Immunizing dose is 10 μ g/
At every turn/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 epitope polypeptide of the defined epitope on native protein or the gene recombinant protein;
2) said one or more 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 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 true CN1259524A (en) | 2000-07-12 |
| CN1152053C CN1152053C (en) | 2004-06-02 |
Family
ID=5283004
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 |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN1152053C (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109517045A (en) * | 2017-12-19 | 2019-03-26 | 北京泽勤生物医药有限公司 | A kind of low pH of modified is inserted into peptide |
| CN110272502A (en) * | 2019-07-12 | 2019-09-24 | 深圳市亚辉龙生物科技股份有限公司 | The hybridoma and preparation method, monoclonal antibody and application of immunogene, the anti-cardiac muscle troponin I monoclonal antibody of secretion |
-
1999
- 1999-11-12 CN CNB991238087A patent/CN1152053C/en not_active Expired - Fee Related
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109517045A (en) * | 2017-12-19 | 2019-03-26 | 北京泽勤生物医药有限公司 | A kind of low pH of modified is inserted into peptide |
| CN110790829A (en) * | 2017-12-19 | 2020-02-14 | 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 |
| 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 |
| CN110272502A (en) * | 2019-07-12 | 2019-09-24 | 深圳市亚辉龙生物科技股份有限公司 | The hybridoma and preparation method, monoclonal antibody and application of immunogene, the anti-cardiac muscle troponin I monoclonal antibody of secretion |
Also Published As
| Publication number | Publication date |
|---|---|
| CN1152053C (en) | 2004-06-02 |
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