Human source anti-hepatitis A virus gene engineering antibody from CHO cell

A technology of genetic engineering antibody and hepatitis A virus, which is applied in the direction of genetic engineering, antiviral immunoglobulin, antibody, etc., and can solve problems such as immune diseases and antibody failure

Inactive Publication Date: 2005-04-13
中国疾病预防控制中心病毒病预防控制所
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The use of pure mouse monoclonal antibodies to prevent and treat viral diseases has not been approved internationally. The biggest disadvantage of using mouse antibodie

Method used

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  • Human source anti-hepatitis A virus gene engineering antibody from CHO cell
  • Human source anti-hepatitis A virus gene engineering antibody from CHO cell
  • Human source anti-hepatitis A virus gene engineering antibody from CHO cell

Examples

Experimental program
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Effect test

Embodiment 1

[0034]Embodiment 1 is the sequence characteristics of the human source anti-hepatitis A virus genetic engineering antibody gene produced by CHO cells; embodiment 2-4 is the cloning preparation method of the human source anti hepatitis A virus genetic engineering antibody produced by CHO cells; embodiment 5 is CHO cell The protein characteristics of the produced human-sourced anti-hepatitis A virus genetically engineered antibody; Examples 6-9 are the functional characteristics of the human-sourced anti-hepatitis A virus genetically engineered antibody produced by CHO cells.

example 1

[0035] Example 1, the sequence characteristics of the human anti-hepatitis A virus genetically engineered antibody gene produced by CHO cells: its variable region gene composition characteristics are specific light chain and heavy chain genes, derived from the human anti-hepatitis A virus antibody gene library Specific enrichment screening and expression, the corresponding three CDR region sequences are novel sequences unique to this antibody, and the ChHAIgG16 light chain and heavy chain variable region gene sequences have been stated in the patent application CN1316437A. ChHAIgG78 light chain variable region CDR1, CDR2 and CDR3 sequences are "RASQSVSSSYLA", "GASSRAT" and "QQYGSSPLT", respectively. Its complete nucleotide sequence is as follows:

[0036] GAGCTCACGCAGTCTCCA GGCACCCTGTCTTTGTCTCCAGG GAAAGAGCC

[0037] ACC CTC TCC TGC AGG GCCAGTCAGGTGT AGCAGCAGCTACTTA GCC

[0038] TGGTACCAGCAGAAACCTGGCCAGGCTCCCAGGCTCCTCATCTATGGTGCATC

[0039] CAGCAGGGCCACTGGC ATCCCAGACAGGTTCAG...

example 2

[0052] Example 2, Cloning and recombination of the human anti-hepatitis A virus genetically engineered antibody gene produced by CHO cells: the anti-hepatitis A virus neutralizing antibody Fab gene obtained through the phage surface display system was cloned into the human antibody high-efficiency expression expression vector invented by our laboratory middle. Among them, the ChHAIgG16 and ChHAIgG78 light chain variable region genes were respectively cloned into the vector VK-dhfrl with the EcoRV and XhoI endonuclease sites (this vector has been stated in the patent application with the patent publication number of CN1363682), placed in the EF-1α promoter Under the control of the child, the expression vectors 16VL-dhfr and 78VK-dhfr were obtained. The heavy chain variable region genes of ChHAIgG16 and ChHAIgG78 were respectively cloned into the vector VH-dhfr1 with BssHII and BstE II (this vector has been stated in the patent application CN1363682), and the expression vectors ...

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Abstract

The human-derived anti-Hepatitis A virus genetically engineered antibody produced by CHO cells includes two strains of human-derived anti-Hepatitis A virus neutralizing genetically engineered antibody CHO (Chinese hamster ovary cell) production cell line and the production of human-derived anti-Hepatitis A virus neutralizing for prevention and treatment Genetically engineered antibody preparations. Its product features are fully human antibodies IgG1λ and IgG1κ derived from phage human antibody library screening and human IgG Fc gene recombination in Chinese hamster ovary (CHO) cells stably expressing cells, named ChHAIgG16 and ChHAIgG78 respectively. The molecular weight is about 150kd. The cocktail-type antibody preparation composed of ChHAIgG16 and ChHAIgG78 mixed at a ratio of 3:1 is named HAMaxTMM. Utilizing its functional characteristics of highly neutralizing hepatitis A virus infection and the molecular structure characteristics of whole antibodies similar to natural antibody molecules, it is expected to replace blood product gamma globulin in clinical practice for the prevention and treatment of hepatitis A caused by hepatitis A virus infection .

Description

1. Technical field [0001] The invention belongs to the technical fields of bioengineering and biopharmaceuticals. 2. Background technology [0002] The invention includes two strains of human anti-hepatitis A virus neutralizing genetic engineering antibodies. respectively by China Production in murine ovary cell (CHO) production cell lines. The antibody and its preparation can be used to prepare medicines or reagents for diagnosing, preventing and treating hepatitis A caused by hepatitis A virus infection. [0003] The number of deaths caused by infectious diseases still ranks first in the world's death population, and viral hepatitis is an infectious disease that seriously endangers the health of our people. Among the infectious diseases notifiable by law, the acute and chronic hepatitis caused by various hepatitis viruses ranks first in the morbidity and mortality of all infectious diseases in my country. Hepatitis A (Hepatitis A) is an acute infectious disease caused...

Claims

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Application Information

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IPC IPC(8): A61K39/42A61K48/00C07K16/08C12N5/10C12N15/13C12N15/85
Inventor 梁米芳曹经媛李川李德新
Owner 中国疾病预防控制中心病毒病预防控制所
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