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Fuse protein of specific antibody N1M1 and N2M2 and its coding gene

A bispecific antibody and fusion protein technology, applied in the direction of anti-receptor/cell surface antigen/cell surface determinant immunoglobulin, anti-animal/human immunoglobulin, DNA/RNA fragments, etc., can solve the preparation process Complexity, various products, poor stability of double hybridoma cell lines, etc.

Inactive Publication Date: 2006-02-22
SHENYANG INST OF APPLIED ECOLOGY - CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

But the disadvantage is: the newly prepared BsAb is easily degraded and eliminated in the body
[0007] 2. Hybrid hybridoma preparation method: The hybrid hybridoma method to prepare bispecific antibodies is based on traditional monoclonal antibody technology, and hybridoma cells secreting two antibodies are fused to produce hybrid hybridomas that stably secrete BsAb ; The double antibody prepared by this method has high biological activity and relatively stable antibody structure, but the stability of the double hybridoma cell line is poor in the passage process, and the preparation process is complicated and the yield is low. Ten kinds of antibodies are formed after linkage, only one of which is a biologically active BsAb, and the products are numerous and difficult to purify

Method used

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  • Fuse protein of specific antibody N1M1 and N2M2 and its coding gene
  • Fuse protein of specific antibody N1M1 and N2M2 and its coding gene
  • Fuse protein of specific antibody N1M1 and N2M2 and its coding gene

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] A single-chain antibody gene NM3E2 of a human anti-CD16 molecule has the base sequence of SEQ ID No: 1 in the sequence table after being amplified by primers.

[0034] TAACCATG GAGGTGCAGCTGGTGGAGTCTGGGGGAGGTGTGGTACGGCCTGGGGGGTCCCTGAG

[0035] ACTCTCCTGTGCAGCCTCTGGATTCACCTTTGATGATTATGGCATGAGCTGGGTCCGCCAAGCT

[0036] CCAGGGAAGGGGCTGGAGTGGGTCTCTGGTATTAATTGGAATGGTGGTAGCACAGGTTATGCAG

[0037] ACTCTGTGAAGGGCCGATTCACCATTCTCCAGAGACAACGCCAAGAACTCCCTGTATCTGCAAAT

[0038]GAACAGTCTGAGAGCCGAGGACACGGCCGTGTATTACTGTGCAAGAGGGAGGTCTCTGCTTTTT

[0039] GACTATTGGGGCCAAGGTACCCTGGTCACCGTCTCTAGAGGTGGAGGCGGTTCAGGCGGAGGTG

[0040] GCTCTGGCGGTGGCGGATCGTCTGAGCTGACTCAGGACCCTGCTGTGTCTGTGGCCTTGGGACA

[0041] GACAGTCAGGATCACATGCCAAGGAGACAGCCTCAGAAGCTATTATGCAAGCTGGTACCAGCAG

[0042] AAGCCAGGACAGGCCCCTGTACTTGTCATCTATGGTAAAAACAACCGGCCCTCAGGGATCCCAG

[0043] ACCGATTCTCTGGCTCCAGCTCAGGAAACACAGCTTCCTTGACCATCACTGGGGCTCAGGCGGA

[0044] AGATGAGGCTGACTATTACTGTAACTCCCGGGACAGCAGTGGTAACCATGAGGTATTCGGCGGA ...

Embodiment 2

[0061] A mouse-derived anti-melanoma surface antigen GD2 single-chain antibody gene M-ScFv is derived from hybridoma HB8759, and has the base sequence of SEQ ID No: 2 in the sequence table after amplification by primers.

[0062] TCAGGTGGAGGCGGTTCT TCAGGAGGAGGCTTGGTGCAACCTGGAGGATCCATGAAACTCTCCT

[0063] GTGTTGTCTCTGGATTCACTTTCAGTAATTACTGGATGAACTGGGTCCGCCAGTCTCCAGAGAA

[0064] GGGGCTTGAGTGGATTGCAGAAATTAGATTGAAATCCAATAATTTTGCAAGATATTATGCGGAG

[0065] TCTGTGAAAGGGAGGTTCACCATCTCAAGAGATGATTCCAAAGGTAGTGTCTACCTGCAAATGA

[0066] TCAACCTAAGAGCTGAAGATACTGGCATTTATTACTGTACCAGTTATGGTAACTACGTTGGGCA

[0067] CTATTTTGACCACTGGGGCCAAGGGACCACGGTCACCGTCTCCTCAGGTGGAGGCGGTTCAGGC

[0068] GGAGGTGGCTCTGGCGGTGGCGGATCGGACATTGAGCTCACCCAGTCTCCAAAATTCATGTCCA

[0069] CATCAGTAGGAGACAGGGTCAGCGTCACCTGCAAGGCCAGTCAGAATGTGGATACTAATGTAGC

[0070] CTGGTATCAACAAAAACCAGGGCAATCTCCTGAACCACTGCTTTTCTCGGCATCCTACCGTTAC

[0071] ACTGGAGTCCCTGATCGCTTCACAGGCAGTGGATCTGGGACAGATTTCACTTCTCACCATCAGCA

[0072] ATGTGCAGTC...

Embodiment 3

[0090] The fusion gene of the bispecific antibody N1M1 has the base sequence of SEQ ID No: 3 in the sequence table

[0091] ATGGAGGTGCAGCTGGTGGAGTCTGGGGGAGGTGTGGTACGGCCTGGGGGGTCCCTGAGACTCT

[0092] CCTGTGCAGCCTCTGGATTCACCTTTGATGATTATGGCATGAGCTGGGTCCGCCAAGCTCCAGG

[0093] GAAGGGGCTGGAGTGGGTCTCTGGTATTAATTGGAATGGTGGTAGCACAGGTTATGCAGACTCT

[0094] GTGAAGGGCCGATTCACCATCTCCAGAGACAACGCCAAGAACTCCCTGTATCTGCAAATGAACA

[0095] GTCTGAGAGCCGAGGACACGGCCGTGTATTACTGTGCAAGAGGGAGGTCTCTGCTTTTTGACTA

[0096] TTGGGGCCAAGGTACCCTGGTCACCGTCTCTAGAGGTGGAGGCGGTTCAGGCGGAGGTGGCTCT

[0097] GGCGGTGGCGGATCGTCTGAGCTGACTCAGGACCCTGCTGTGTCTGTGGCCTTGGGACAGACAG

[0098] TCAGGATCACATGCCAAGGAGACAGCCTCAGAAGCTATTATGCAAGCTGGTACCAGCAGAAGCC

[0099]AGGACAGGCCCCTGTACTTGTCATCTATGGTAAAAACAACCGGCCCTCAGGGATCCCAGACCGA

[0100] TTCTCTGGCTCCAGCTCAGGAAACACAGCTTCCTTGACCATCACTGGGGCTCAGGCGGAAGATG

[0101] AGGCTGACTATTACTGTAACTCCCGGGACAGCAGTGGTAACCATGAGGTATTCGGCGGAGGGAC

[0102] CAAGCTGACCGTCCTAGGTTCAGGTGGAGGCGGTTCTCAGGTGAAG...

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Abstract

Disclosed is a fusion protein of specific antibody N1M1 and N2M2 and its coding gene, wherein the double specific antibody N1M1 fusion protein has amino acid sequence of SEQ ID No:2 in the sequence table, its coding gene has amino acid sequence of SEQ ID No:3 in the sequence table, and the double specific antibody N2M2 fusion protein has amino acid sequence of SEQ ID No:8 in the sequence table, its coding gene has amino acid sequence of SEQ ID No:7 in the sequence table.

Description

technical field [0001] The invention belongs to the field of pharmacy, specifically a novel anti-CD16 / anti-melanoma bispecific antibody N1M1 and N2M2 fusion protein and its coding gene (expression and application in Escherichia coli). Background technique [0002] For a long time, the clinical treatment of tumors has been plagued by three major problems: conventional radiotherapy and chemotherapy regimens are poorly selective; tumor cells develop drug resistance; tumor micrometastases occur. And tumor immunotherapy undoubtedly provides a new way to solve these problems. [0003] Since the successful establishment of monoclonal antibody technology by Kohler and Milstein in the mid-1970s, this therapy has been extensively studied and discussed. However, the efficacy of monoclonal antibodies in the treatment of solid tumors is not obvious. Therefore, bispecific antibodies (Bispecificantibody, BsAb) that can target drugs and mediate immune cells have become one of the hot spot...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/62C07K16/18C07K16/30
Inventor 吴文芳倪剑锋孙静吕安国
Owner SHENYANG INST OF APPLIED ECOLOGY - CHINESE ACAD OF SCI
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