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

A chimeric antigen receptor and targeting technology, applied in the field of medical biology, can solve problems such as limitations and lack of relevant research on a variety of EGFR-positive solid tumors, and achieve efficient metastasis, wide application range, and highly tissue-specific expression Effect

Inactive Publication Date: 2019-12-03
SHENZHEN BINDEBIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the current application of CART technology is still limited to hematological tumors, and there is no relevant research on the application of various EGFR-positive solid tumors.

Method used

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

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0052] A method for preparing chimeric antigen receptor T cells targeting EGFR, comprising the following steps:

[0053] (1) Prepare the gene sequence of chimeric antigen receptor CAR-EGFR targeting EGFR

[0054] The coding gene sequence of the chimeric antigen receptor CAR-EGFR targeting EGFR is prepared by PCR, and the coding gene of the CAR-EGFR includes the nucleotide sequence shown in SEQ ID NO: 3;

[0055] (2) Construction of pWPXLd-CAR-EGFR recombinant plasmid

[0056] The gene encoding CAR-EGFR was inserted between the BamHI and EcoRI restriction sites of the pWPXLD vector, and behind the EF1α of the pWPXLD vector, EF1α was used as the promoter. When the CAR-EGFR coding gene is inserted into the pWPXLD vector, a start codon (such as ATG) is added to the 5' end of the CAR-EGFR coding gene to connect to the BamHI restriction site in the pWPXLD vector, and the 3' end is also A stop codon (such as TAA) is added to connect with the EcoRI restriction site in the pWPXLD vec...

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Abstract

The invention provides an EGFR-targeting chimeric antigen receptor CAR-EGFR. The CAR-EGFR comprises an amino acid sequence as shown in SEQ ID NO:1. The invention further provides a chimeric antigen receptor T cell including the CAR-EGFR. The EGFR-targeting chimeric antigen receptor CAR-EGFR can specifically target EGFR positive tumor cells and activate T cells to exert cellular immunity effects torealize efficient and specific killing on the EGFR positive tumor cells, has persistent cell viability and lethality, and cannot damage normal cells. The invention further provides a preparation method and application of the EGFR targeted chimeric antigen receptor T cell.

Description

technical field [0001] The invention relates to the field of medical biology, in particular to a chimeric antigen receptor targeting EGFR, a chimeric antigen receptor T cell and a preparation method and application thereof. Background technique [0002] Cancer prevention and treatment is difficult and the disease burden is heavy, and it has become one of the major public health problems that seriously endanger human health. Epidermal growth factor receptor (EGFR) is a receptor for epidermal growth factor (EGF) cell proliferation and signal transduction, and belongs to the tyrosine kinase type I receptor subfamily. Studies have shown that there are high or abnormal expressions of EGFR in many solid tumors. EGFR is related to tumor cell proliferation, angiogenesis, tumor invasion, metastasis and inhibition of apoptosis. Therefore, in tumor therapy, EGFR can become a very important drug target. [0003] Chimeric Antigen Receptor–Modified T Cells (CAR-T), as one of the latest...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K19/00C12N15/62C12N15/867C12N5/10A61K35/17A61P35/00
CPCA61K35/17A61P35/00C07K14/7051C07K14/70578C07K16/2863C07K2319/00C07K2319/33C07K2319/74C12N5/0636C12N15/86C12N2510/00C12N2740/15043C12N2800/107
Inventor 万晓春刘绿艳唐超李欣
Owner SHENZHEN BINDEBIOTECH CO LTD
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