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Anti-CD24 (cluster of differentiation 24) humanized antibody fusion protein

A technology of humanized antibody and fusion protein, which is applied in the field of bioengineering in Ming Dynasty

Inactive Publication Date: 2017-10-03
CHINA PHARM UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

So far, there are few reports on fusion proteins related to NKG2D ligand MICA molecules

Method used

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  • Anti-CD24 (cluster of differentiation 24) humanized antibody fusion protein
  • Anti-CD24 (cluster of differentiation 24) humanized antibody fusion protein
  • Anti-CD24 (cluster of differentiation 24) humanized antibody fusion protein

Examples

Experimental program
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Effect test

Embodiment 1

[0029] Example 1 Construction of CD24 antibody fusion protein hG7-MICA

[0030] Using hG7 heavy and light chain genes, MICA gene, pCA puro, and pMH3 plasmids as templates, primers were designed and synthesized for PCR amplification. The fusion protein hG7-MICA was constructed based on the hG7 heavy chain gene H, and the complete H-MICA gene was obtained by overlapping PCR extension amplification. The PCR product was detected by 1.0% agarose gel electrophoresis, and the target gene was recovered by the agarose gel extraction kit. The final products of PCR amplification and plasmids pCApuro and pMH3 were double-digested with restriction endonucleases, and the digested products were gel-recovered and then ligated with T4 ligase overnight at 16°C. After ligation, transform Escherichia coli DH5α competent, smear the plate, pick out the single clone, double enzyme digestion and sequence identification the next day.

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PUM

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Abstract

The invention belongs to the technical field of genetic engineering antibodies, in particular to a preparation method and application of a CD24 antibody fusion protein. The invention utilizes genetic engineering technology to link the CD24 humanized antibody hG7S with the extracellular 1-3 region of the MICA molecule through the G4S flexible peptide, and the mammalian eukaryotic expression system stably expresses hG7S-MICA; the CD24 antibody fusion protein hG7S-MICA can simultaneously Combined with CD24 molecules on the surface of tumor cells and the activated receptor NKG2D on the surface of NK cells, remodeling the immune surveillance effect of NK cells induced by NKG2D pathway on CD24+ tumor cells, and finally achieving the goal of activating the body's own immune system to effectively kill CD24+ tumor cells Purpose.

Description

technical field [0001] The invention belongs to the field of bioengineering, and in particular relates to a new CD24 humanized antibody fusion protein hG7-MICA that can specifically bind to the leukocyte differentiation antigen CD24 (cluster of differentiation 24) and NKG2D receptors at the same time, and uses hG7 target The tropism specifically displays MICA on the surface of tumor cells, enhances the antibody-dependent cell-mediated cytotoxicity (ADCC) of NK cells on human breast cancer cells, and avoids the down-regulation and shedding of MICA molecules on the cell surface. Tumor immune escape. The design of hG7-MICA opens up a new tumor immunotherapy strategy, which is a bifunctional antibody fusion protein that specifically remodels the immune surveillance function of NK cells on CD24+ tumor cells. Background technique [0002] The morbidity and mortality of malignant tumors in the world have been on the rise. Tumor immune escape refers to the shedding of tumor cell s...

Claims

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Application Information

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IPC IPC(8): C07K19/00A61K39/00A61K47/68A61P35/00
CPCC07K16/2896A61K39/00C07K14/70539C07K2319/00C07K2319/33
Inventor 王旻张娟孙福谋杜晓典
Owner CHINA PHARM UNIV
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