CN1232039A - Genetic engineering double specific antibody and its use - Google Patents
Genetic engineering double specific antibody and its use Download PDFInfo
- Publication number
- CN1232039A CN1232039A CN 99103517 CN99103517A CN1232039A CN 1232039 A CN1232039 A CN 1232039A CN 99103517 CN99103517 CN 99103517 CN 99103517 A CN99103517 A CN 99103517A CN 1232039 A CN1232039 A CN 1232039A
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- China
- Prior art keywords
- hepatitis
- surface antigen
- genetic engineering
- antibody
- specific antibody
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- 238000010353 genetic engineering Methods 0.000 title claims description 8
- 239000000427 antigen Substances 0.000 claims abstract description 18
- 108091007433 antigens Proteins 0.000 claims abstract description 18
- 102000036639 antigens Human genes 0.000 claims abstract description 18
- 108090000623 proteins and genes Proteins 0.000 claims abstract description 13
- 210000004369 blood Anatomy 0.000 claims abstract description 8
- 239000008280 blood Substances 0.000 claims abstract description 8
- 208000002672 hepatitis B Diseases 0.000 claims abstract description 8
- 238000000034 method Methods 0.000 claims abstract description 8
- 239000003153 chemical reaction reagent Substances 0.000 claims abstract description 5
- 238000001514 detection method Methods 0.000 claims abstract description 4
- 108090000765 processed proteins & peptides Proteins 0.000 claims abstract description 4
- 239000013604 expression vector Substances 0.000 claims abstract description 3
- 208000006454 hepatitis Diseases 0.000 claims description 10
- 102000004196 processed proteins & peptides Human genes 0.000 claims description 3
- 239000000047 product Substances 0.000 abstract description 3
- 238000004904 shortening Methods 0.000 abstract description 2
- 239000006228 supernatant Substances 0.000 abstract description 2
- 210000003743 erythrocyte Anatomy 0.000 abstract 2
- 238000005215 recombination Methods 0.000 abstract 2
- 230000006798 recombination Effects 0.000 abstract 2
- 230000001580 bacterial effect Effects 0.000 abstract 1
- 230000009977 dual effect Effects 0.000 abstract 1
- 239000007788 liquid Substances 0.000 abstract 1
- 238000002407 reforming Methods 0.000 abstract 1
- 239000012228 culture supernatant Substances 0.000 description 2
- 238000009629 microbiological culture Methods 0.000 description 2
- 238000002360 preparation method Methods 0.000 description 2
- 241000894006 Bacteria Species 0.000 description 1
- 230000000259 anti-tumor effect Effects 0.000 description 1
- 210000000624 ear auricle Anatomy 0.000 description 1
- 230000035931 haemagglutination Effects 0.000 description 1
- 231100000283 hepatitis Toxicity 0.000 description 1
- 238000009434 installation Methods 0.000 description 1
- 230000000968 intestinal effect Effects 0.000 description 1
- 239000003550 marker Substances 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 230000009466 transformation Effects 0.000 description 1
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- Peptides Or Proteins (AREA)
- Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
Abstract
A dual-specific antibody composed of the surface antigens to resist against erythrocyte and hepatitis B, which can be used as the test reagent to detect the surface antigen of hepatitis B in blood, is prepared by recombination technique in gene engineering. The recombination technique includes reforming the surface antigen to resist against erythrocyte and hepatitis B to become hybrid antibody genes by shortening the joining peptide of single-chain antibody, and assembling them in same expression vector. Said gene product can be directly extracted from the supernatant of bacterial culture liquid. Its advantages are low cost, quick detection and simple operation.
Description
The invention discloses a kind of genetic engineering double specific antibody and application thereof, particularly a kind of genetic engineering double specific antibody and application of forming by anti-erythrocyte and anti-hepatitis b surface antigen thereof.
At present, the bispecific antibody of forming by antitumor cell and anti-immunocyte surface marker CD3 of the method preparation of professor Winter that known bispecific antibody mainly contains univ cambridge uk in 1993 by shortening the single-chain antibody connection peptides, be called two special Diabody, it mainly acts on antineoplaston clinically.Still do not have record and use by genetic engineering double specific antibody and application thereof that anti-erythrocyte and anti-hepatitis b surface antigen are formed.
The object of the present invention is to provide a kind of genetic engineering double specific antibody and application of forming by anti-erythrocyte and anti-hepatitis b surface antigen thereof.
The object of the present invention is achieved like this:
A. bispecific antibody gene:
The present invention utilizes the genetically engineered recombinant technology by the method that shortens the single-chain antibody connection peptides anti-erythrocyte and anti-hepatitis b surface antigen to be transformed into the hybrid antibody gene, is assembled in that (reference of carrier PCOMb3H is CarlosF.Barbas III andJurgen Wagner Synthetic Human Antibodies:Selecting andEvolving Functional Proteins METHODS:A Companion toMethods in Enzymology 1995 in the PCOMb3H expression vector; 8:94-103), constitute anti-erythrocyte and anti-hepatitis b surface antigen bispecific antibody; The bispecific antibody gene order is
GCCGAGCTCA?CCCAGTCTCC?ATCCTTCCTG?TCTGCATCTA
TAGGAGACAG?CGTCACCATC?ACTTGCCGGA?CCAGTCAGGA
CATTGATAAT?TATTTAGCCT?GGTATCAGCA?AAAACCAGGA
AAAGCCCCTA?AGCTCCTGGT?CTATTCTGCA?TCCGCCTTGC
AAGGTGGGGT?CCCATCAAGG?TTCAGCGGCA?GTGGATCTAG
GACAGATTTC?ACTCTCACAA?TCAACAGCCT?GCAGCCTGAA
GATTCTGCAA?CTTATTACTG?TCAACAGCTT?GGAAGTTACC
CACTCACTTT?CGGCGGAGGG?ACCAAGGTGG?AGATCAAACG
ATCTAGAGGT?GGTGGATCCG?AGGTGCAGCT?GCTCGAGCTT
GCGGAGTCTG?GGGGAGGCTT?AGTGAAGCCT?GGAGGGTCCC
TGAAACTCTC?CTGTACAACC?TCTGGATTCG?CTTTCAGTAG
TTATGACATG?TCTTGGGTTC?GCCAGACTCC?GGAGAAGAGG
CTGGAGTGGG?TCGCATACAT?TAGTAGTGCT?GGTGGTAGTA
TTTATTATTC?AGACACTGTG?AAGGACCGCT?TCACCATCTC
CAGAGACAAT?GTCAAAAACA?CCCTGTACCT?GCAAATGAAC
AGTCTGAATT?CTGAGGACAC?AGCCATATAT?TACTGTACAA
GACACGGGCT?CGGCTCCTAC?TATGTTATGG?ACTACTGGGG
TCAAGGAACC?TCAGTCACCG?TCTCCTCAAC?TAGTGAACAA
AAACTCATCT?CAGAAGAAGA?TCTGGCTAGC?CATCACCATC
ACCATCACTA?AGTCGACAGG?AGGAATTTAA?AATGAAAAAG
ACAGCTATCG?CGATTGCAGT?GGCACTGGCT?GGTTTCGCTA
CCGTGGCCCA?GGCGGCCGAG?CTCCAGATGA?CCCAGTCTCC
AGCCTCCCTA?TCTGCATCTG?TGGGAGAAAC?TGTCACCATC
ACATGTCGAG?CAAGTGAGAA?TATTTACAGT?TATTTAGCAT
GGTATCAGCA?GAAACAGGGA?AAATCTCCTC?AGCTCCTGGT
CTATAATGCA?AAAACCTTAG?CAGAAGGTGT?GCCATCAAGG
TTCAGTGGCA?GTGGATCAGG?CACACAGTTT?TCTCTGAAGA
TCAACAGCCT?GCAGCCTGAA?GATTTTGGGA?GTTATTACTG
TCAACATCAT?TATGGTACTC?GTCTCACGTT?CGGTGCTGGG
ACCAAGCTGG?AGCTGAAACG?GTCTAGAGGT?GGTGGATCCG
AGGTGCAGGT?GCTCGAGGAG?TCTGGGGGAG?GCTTGGTACA
GGCTGGGGGG?TCCCTGAGAC?TCTCCTGTGC?AATCTCTGGA
TTCACCTTTA?GTGACTATGC?CGTGAGTTGG?GTCCGCCAGG
CTCCTGGGAA?GGGGCTGGAG?TGGGTCTCAA?CCATCAGTGG
CAGTGCTGGA?AACACATACT?ACGCAGACTC?CGTGAAGGGC
CGATTGATCA?TCTCCAGAGA?CACTTCCAGG?AACATGCTAT
TTCTGCAAAT?GAACAGCCTG?AGAGCCGAGG?ACACGGCCGT
ATATTACTGT?GCGAAAGTCA?AGATGGTTCG?GGGTGGCTAC
TGGTTCTACG?GTATGGACGT?CTGGGGCCAA?GGGACCGCGG
TCAGAGTCTC?CTCA；
B. use:
Bispecific antibody gene transformation intestinal bacteria justacrine is expressed, and centrifugal collection microbial culture supernatant promptly contains anti-erythrocyte and anti-hepatitis b surface antigen bispecific antibody in the supernatant, can detect hepatitis B surface antigen in the blood as the hepatitis B surface antigen detection reagent.
The bispecific antibody that adopts the two specific genes of anti-erythrocyte and anti-hepatitis b surface antigen to be prepared from because gene engineering product can directly obtain, has reduced the preparation cost of product from the microbial culture supernatant; In testing process, do not need to use specific installation, only need gather not tip blood, place in room temperature and just can finish detection in 1～2 minute, be not subjected to the influence in time and geographical position, have easy and simple to handle, advantage efficiently.
The method of using bispecific antibody to detect hepatitis B surface antigen in the blood is to drip bispecific antibody reagent 50ml on slide, gather not tip blood (referring to blood or ear-lobe blood) 20ml and reagent mixing, placed 1～2 minute in room temperature, slide is shaken the aggegation situation of observing, result of determination, the hemagglutination person is a hepatitis b surface antigen positive.
Claims (2)
1. genetic engineering double specific antibody and application thereof, utilize the genetically engineered recombinant technology anti-erythrocyte and anti-hepatitis b surface antigen to be transformed into the hybrid antibody gene by the method that shortens the single-chain antibody connection peptides, be assembled in the PCOMb3H expression vector, constitute anti-erythrocyte and anti-hepatitis b surface antigen bispecific antibody gene; It is characterized in that: the bispecific antibody gene order is
GCCGAGCTCA?CCCAGTCTCC?ATCCTTCCTG?TCTGCATCTA
TAGGAGACAG?CGTCACCATC?ACTTGCCGGA?CCAGTCAGGA
CATTGATAAT?TATTTAGCCT?GGTATCAGCA?AAAACCAGGA
AAAGCCCCTA?AGCTCCTGGT?CTATTCTGCA?TCCGCCTTGC
AAGGTGGGGT?CCCATCAAGG?TTCAGCGGCA?GTGGATCTAG
GACAGATTTC?ACTCTCACAA?TCAACAGCCT?GCAGCCTGAA
GATTCTGCAA?CTTATTACTG?TCAACAGCTT?GGAAGTTACC
CACTCACTTT?CGGCGGAGGG?ACCAAGGTGG?AGATCAAACG
ATCTAGAGGT?GGTGGATCCG?AGGTGCAGCT?GCTCGAGCTT
GCGGAGTCTG?GGGGAGGCTT?AGTGAAGCCT?GGAGGGTCCC
TGAAACTCTC?CTGTACAACC?TCTGGATTCG?CTTTCAGTAG
TTATGACATG?TCTTGGGTTC?GCCAGACTCC?GGAGAAGAGG
CTGGAGTGGG?TCGCATACAT?TAGTAGTGCT?GGTGGTAGTA
TTTATTATTC?AGACACTGTG?AAGGACCGCT?TCACCATCTC
CAGAGACAAT?GTCAAAAACA?CCCTGTACCT?GCAAATGAAC
AGTCTGAATT?CTGAGGACAC?AGCCATATAT?TACTGTACAA
GACACGGGCT?CGGCTCCTAC?TATGTTATGG?ACTACTGGGG
TCAAGGAACC?TCAGTCACCG?TCTCCTCAAC?TAGTGAACAA
AAACTCATCT?CAGAAGAAGA?TCTGGCTAGC?CATCACCATC
ACCATCACTA?AGTCGACAGG?AGGAATTTAA?AATGAAAAAG
ACAGCTATCG?CGATTGCAGT?GGCACTGGCT?GGTTTCGCTA
CCGTGGCCCA?GGCGGCCGAG?CTCCAGATGA?CCCAGTCTCC
AGCCTCCCTA?TCTGCATCTG?TGGGAGAAAC?TGTCACCATC
ACATGTCGAG?CAAGTGAGAA?TATTTACAGT?TATTTAGCAT
GGTATCAGCA?GAAACAGGGA?AAATCTCCTC?AGCTCCTGGT
CTATAATGCA?AAAACCTTAG?CAGAAGGTGT?GCCATCAAGG
TTCAGTGGCA?GTGGATCAGG?CACACAGTTT?TCTCTGAAGA
TCAACAGCCT?GCAGCCTGAA?GATTTTGGGA?GTTATTACTG
TCAACATCAT?TATGGTACTC?GTCTCACGTT?CGGTGCTGGG
ACCAAGCTGG?AGCTGAAACG?GTCTAGAGGT?GGTGGATCCG
AGGTGCAGCT?GCTCGAGGAG?TCTGGGGGAG?GCTTGGTACA
GCCTGGGGGG?TCCCTGAGAC?TCTCCTGTGC?AATCTCTGGA
TTCACCTTTA?GTGACTATGC?CGTGAGTTGG?GTCCGCCAGG
CTCCTGGGAA?GGGGCTGGAG?TGGGTCTCAA?CCATCAGTGG
CAGTGCTGGA?AACACATACT?ACGCAGACTC?CGTGAAGGGC
CGATTGATCA?TCTCCAGAGA?CACTTCCAGG?AACATGCTAT
TTCTGCAAAT?GAACAGCCTG?AGAGCCGAGG?ACACGGCCGT
ATATTACTGT?GCGAAAGTCA?AGATGGTTCG?GGGTGGCTAC
TGGTTCTACG?GTATGGACGT?CTGGGGCCAA?GGGACCGCGG
TCAGAGTCTC?CTCA；
2. genetic engineering double specific antibody and application thereof is characterized in that: bispecific antibody can detect hepatitis B surface antigen in the blood as the hepatitis B surface antigen detection reagent.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 99103517 CN1232039A (en) | 1999-04-02 | 1999-04-02 | Genetic engineering double specific antibody and its use |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 99103517 CN1232039A (en) | 1999-04-02 | 1999-04-02 | Genetic engineering double specific antibody and its use |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN1232039A true CN1232039A (en) | 1999-10-20 |
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN 99103517 Pending CN1232039A (en) | 1999-04-02 | 1999-04-02 | Genetic engineering double specific antibody and its use |
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Cited By (15)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101668771A (en) * | 2007-03-12 | 2010-03-10 | 艾斯巴技术股份公司 | Sequence-based engineering and optimization of single-chain antibodies |
| CN103068846A (en) * | 2010-08-24 | 2013-04-24 | 弗·哈夫曼-拉罗切有限公司 | Bispecific antibodies comprising disulfide bond-stabilized Fv fragments |
| US9688758B2 (en) | 2012-02-10 | 2017-06-27 | Genentech, Inc. | Single-chain antibodies and other heteromultimers |
| US9890204B2 (en) | 2009-04-07 | 2018-02-13 | Hoffmann-La Roche Inc. | Trivalent, bispecific antibodies |
| US9994646B2 (en) | 2009-09-16 | 2018-06-12 | Genentech, Inc. | Coiled coil and/or tether containing protein complexes and uses thereof |
| US10106612B2 (en) | 2012-06-27 | 2018-10-23 | Hoffmann-La Roche Inc. | Method for selection and production of tailor-made highly selective and multi-specific targeting entities containing at least two different binding entities and uses thereof |
| US10106600B2 (en) | 2010-03-26 | 2018-10-23 | Roche Glycart Ag | Bispecific antibodies |
| US10323099B2 (en) | 2013-10-11 | 2019-06-18 | Hoffmann-La Roche Inc. | Multispecific domain exchanged common variable light chain antibodies |
| US10611825B2 (en) | 2011-02-28 | 2020-04-07 | Hoffmann La-Roche Inc. | Monovalent antigen binding proteins |
| US10633457B2 (en) | 2014-12-03 | 2020-04-28 | Hoffmann-La Roche Inc. | Multispecific antibodies |
| US10640555B2 (en) | 2009-06-16 | 2020-05-05 | Hoffmann-La Roche Inc. | Bispecific antigen binding proteins |
| US10793621B2 (en) | 2011-02-28 | 2020-10-06 | Hoffmann-La Roche Inc. | Nucleic acid encoding dual Fc antigen binding proteins |
| US10927163B2 (en) | 2007-12-21 | 2021-02-23 | Hoffmann-La Roche, Inc. | Bivalent, bispecific antibodies |
| US11421022B2 (en) | 2012-06-27 | 2022-08-23 | Hoffmann-La Roche Inc. | Method for making antibody Fc-region conjugates comprising at least one binding entity that specifically binds to a target and uses thereof |
| US11618790B2 (en) | 2010-12-23 | 2023-04-04 | Hoffmann-La Roche Inc. | Polypeptide-polynucleotide-complex and its use in targeted effector moiety delivery |
-
1999
- 1999-04-02 CN CN 99103517 patent/CN1232039A/en active Pending
Cited By (21)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101668771B (en) * | 2007-03-12 | 2013-08-21 | 艾斯巴技术,爱尔康生物医药研究装置有限责任公司 | Sequence-based engineering and optimization of single-chain antibodies |
| CN101668771A (en) * | 2007-03-12 | 2010-03-10 | 艾斯巴技术股份公司 | Sequence-based engineering and optimization of single-chain antibodies |
| US10927163B2 (en) | 2007-12-21 | 2021-02-23 | Hoffmann-La Roche, Inc. | Bivalent, bispecific antibodies |
| US11993642B2 (en) | 2009-04-07 | 2024-05-28 | Hoffmann-La Roche Inc. | Trivalent, bispecific antibodies |
| US9890204B2 (en) | 2009-04-07 | 2018-02-13 | Hoffmann-La Roche Inc. | Trivalent, bispecific antibodies |
| US10640555B2 (en) | 2009-06-16 | 2020-05-05 | Hoffmann-La Roche Inc. | Bispecific antigen binding proteins |
| US11673945B2 (en) | 2009-06-16 | 2023-06-13 | Hoffmann-La Roche Inc. | Bispecific antigen binding proteins |
| US9994646B2 (en) | 2009-09-16 | 2018-06-12 | Genentech, Inc. | Coiled coil and/or tether containing protein complexes and uses thereof |
| US10106600B2 (en) | 2010-03-26 | 2018-10-23 | Roche Glycart Ag | Bispecific antibodies |
| CN103068846A (en) * | 2010-08-24 | 2013-04-24 | 弗·哈夫曼-拉罗切有限公司 | Bispecific antibodies comprising disulfide bond-stabilized Fv fragments |
| US9879095B2 (en) | 2010-08-24 | 2018-01-30 | Hoffman-La Roche Inc. | Bispecific antibodies comprising a disulfide stabilized-Fv fragment |
| US11618790B2 (en) | 2010-12-23 | 2023-04-04 | Hoffmann-La Roche Inc. | Polypeptide-polynucleotide-complex and its use in targeted effector moiety delivery |
| US10611825B2 (en) | 2011-02-28 | 2020-04-07 | Hoffmann La-Roche Inc. | Monovalent antigen binding proteins |
| US10793621B2 (en) | 2011-02-28 | 2020-10-06 | Hoffmann-La Roche Inc. | Nucleic acid encoding dual Fc antigen binding proteins |
| US9688758B2 (en) | 2012-02-10 | 2017-06-27 | Genentech, Inc. | Single-chain antibodies and other heteromultimers |
| US11407836B2 (en) | 2012-06-27 | 2022-08-09 | Hoffmann-La Roche Inc. | Method for selection and production of tailor-made highly selective and multi-specific targeting entities containing at least two different binding entities and uses thereof |
| US11421022B2 (en) | 2012-06-27 | 2022-08-23 | Hoffmann-La Roche Inc. | Method for making antibody Fc-region conjugates comprising at least one binding entity that specifically binds to a target and uses thereof |
| US10106612B2 (en) | 2012-06-27 | 2018-10-23 | Hoffmann-La Roche Inc. | Method for selection and production of tailor-made highly selective and multi-specific targeting entities containing at least two different binding entities and uses thereof |
| US10323099B2 (en) | 2013-10-11 | 2019-06-18 | Hoffmann-La Roche Inc. | Multispecific domain exchanged common variable light chain antibodies |
| US10633457B2 (en) | 2014-12-03 | 2020-04-28 | Hoffmann-La Roche Inc. | Multispecific antibodies |
| US11999801B2 (en) | 2014-12-03 | 2024-06-04 | Hoffman-La Roche Inc. | Multispecific antibodies |
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