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Trastuzumab mutant IgG and its application

A technology of trastuzumab and mutants, which is applied in the fields of antibodies, applications, and anti-tumor drugs, which can solve the problems of target cell cytotoxicity and achieve the effect of improving ADCC effect

Active Publication Date: 2019-01-11
NANJING GENSCRIPT BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Binding to activating antibody receptors FcγRI, FcγRIIA, and FcγRIIIA will cause an activation response, resulting in cytotoxicity to target cells

Method used

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  • Trastuzumab mutant IgG and its application
  • Trastuzumab mutant IgG and its application
  • Trastuzumab mutant IgG and its application

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0060] 1.1 Construction of eukaryotic expression vector

[0061] The full-length DNA of eight mutant IgGs and the wild-type full-length DNA sequence of Trastuzumab were synthesized respectively. The wild-type heavy chain DNA sequence of Trastuzumab was SEQ ID NO.1, and the heavy chain DNA sequence of mutant 1 IgG was SEQ ID NO.2. The heavy chain DNA sequence of mutant 2IgG is SEQ ID NO.3, the heavy chain DNA sequence of mutant 3IgG is SEQ ID NO.4, the heavy chain DNA sequence of mutant 4IgG is SEQ ID NO.5, and the heavy chain of mutant 5IgG The DNA sequence is SEQ ID NO.6, the heavy chain DNA sequence of mutant 6IgG is SEQ ID NO.7, the heavy chain DNA sequence of mutant 7IgG is SEQ ID NO.8, and the heavy chain DNA sequence of mutant 8IgG is SEQ ID NO. 9. The protein sequences of these clones are SEQ ID10-18. The light chains of Trastuzumab wild type and all mutants described herein are the same, its DNA sequence is SEQ ID19, and its protein sequence is SEQ ID20. These clones...

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Abstract

Belonging to the fields of gene engineering and immunological technologies, the invention discloses trastuzumab mutants IgG and application thereof. By performing point mutation on trastuzumab heavy chain coding DNA, 8 trastuzumab IgG mutants can be obtained, the heavy chain coding DNA sequence of the mutants can be any of SEQ ID NO.2-SEQ ID NO.9. The protein purification yield and trastuzumab obtained by transient expression of each trastuzumab mutant and trastuzumab are both high. Each mutant IgG can improve the ADCC effect of trastuzumab by different degrees without influencing the IgG yield and thermal stability, and can be applied in anticancer drug preparation as trastuzumab.

Description

technical field [0001] The invention belongs to the field of genetic engineering and immune technology, and relates to trastuzumab mutant IgG and application thereof. Background technique [0002] Since OKT3 was first marketed as an antibody drug in 1986, antibody drug development has made incredible progress in the past 25 years. Five of the ten best-selling drugs in 2010 (Humira, Remicade, Enbrel, Rituxan, and Avastin) were antibody drugs or Fc fusion proteins. [0003] Her2 is a member of the EGFR family and was found to be highly expressed in 25-30% of breast cancer patients [1], so Her2 is considered to be an important target for targeted therapy of breast cancer. In 1990, Fendly et al. of Genentech isolated monoclonal antibody 4D5 from a series of mouse monoclonal antibodies, and found that this antibody binds to the extracellular domain of Her2 and inhibits the growth of tumor cells with Her2 overexpression [2]. 4D5 was humanized in 1992, the humanized antibody was ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K16/28C12N15/13C12N15/85C12N5/10A61K39/395A61K48/00A61P35/00
Inventor 张剑冰武术杨帅
Owner NANJING GENSCRIPT BIOTECH CO LTD