Single-chain antibody targeting EGFR, chimeric antigen receptor T cell, preparation method and application thereof

A chimeric antigen receptor and single-chain antibody technology, applied in the field of medical biology, to achieve the effects of promoting amplification, efficient and specific killing, and strong affinity activity

Inactive Publication Date: 2019-06-04
SHENZHEN BINDEBIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there is no preparation and research of chimeric antigen receptor T cells targeting EGFR

Method used

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  • Single-chain antibody targeting EGFR, chimeric antigen receptor T cell, preparation method and application thereof
  • Single-chain antibody targeting EGFR, chimeric antigen receptor T cell, preparation method and application thereof
  • Single-chain antibody targeting EGFR, chimeric antigen receptor T cell, preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0080]A method for preparing chimeric antigen receptor T cells targeting EGFR, comprising the following steps:

[0081] (1) Prepare the gene sequence of chimeric antigen receptor CAR-EGFR targeting EGFR

[0082] The coding genes of the signal peptide, the single-chain antibody targeting EGFR, the CD8α hinge region, the CD8 transmembrane region, the 4-1BB signal region and the CD3ζ signal region are respectively prepared, and the coding genes of the signal peptide are shown in SEQ ID NO: 15 , the coding gene of the single-chain antibody targeting EGFR is shown in SEQ ID NO: 2, the coding gene of the CD8α hinge region is shown in SEQ ID NO: 7, and the coding gene of the CD8 transmembrane region is shown in SEQ ID NO: As shown in ID NO: 9, the coding gene of the 4-1BB signal region is shown in SEQ ID NO: 11, and the coding gene of the CD3ζ signal region is shown in SEQ ID NO: 13.

[0083] The genes encoding the above signal peptide, single-chain antibody targeting EGFR, CD8α hin...

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PUM

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Abstract

The present invention provides a single-chain antibody targeting EGFR, the single-chain antibody targeting EGFR comprises an amino acid sequence shown in SEQ ID NO: 1, and the single-chain antibody targeting EGFR is a humanized single-chain antibody. The present invention also provides a chimeric antigen receptor T cell comprising the single-chain antibody targeting EGFR, wherein the chimeric antigen receptor targeting EGFR can specifically target EGFR, the expansion of T cells in patients is promoted, and the tumor cells can be efficiently and specifically killed. In addition, the chimeric antigen receptor T cell targeting EGFR prepared by humanized single-chain antibody has better cell viability and lethality, and no damage is generated on the normal cells. The invention also provides apreparation method and an application of the chimeric antigen receptor T cell targeting EGFR.

Description

technical field [0001] The invention relates to the field of medical biology, in particular to a single-chain antibody targeting EGFR, a chimeric antigen receptor T cell, and a preparation method and application thereof. Background technique [0002] Human epidermal growth factor receptor (EGFR) is the main receptor for human epidermal growth factor. Upon activation, EGFR undergoes homodimerization or heterodimerization with other members of the human epithelial growth factor receptor family, thereby inducing autophosphorylation of EGFR intracellular structures and activating downstream pathways such as mitogen activation The protein kinase (MAPK) pathway, etc., and mediate cell expansion. Studies have shown that EGFR is a cell surface antigen that is expressed or abnormally expressed in various solid tumor cells such as lung cancer, but weakly expressed in ordinary cells. EGFR is related to the proliferation of tumor cells, tumor invasion, metastasis and inhibition of apo...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/28C12N15/13C07K19/00C12N15/62C12N15/867C12N5/10A61P35/00
Inventor 曾滢杨忠华
Owner SHENZHEN BINDEBIOTECH CO LTD
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