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FGFR4-targeting single-chain antibody, chimeric antigen receptor and chimeric antigen receptor T cell, preparation method of chimeric antigen receptor T cell, and application of FGFR4-targeting single-chain antibody, chimeric antigen receptor and chimeric antigen receptor T cell

A chimeric antigen receptor, FGFR4 technology, applied in the field of medical biology, can solve problems such as drug resistance

Active Publication Date: 2021-05-18
SHENZHEN BINDEBIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

FGFR4 selective inhibitors are basically covalent reversible or irreversible inhibitors based on Cys552. Once this site is mutated, it is likely to lead to drug resistance.
At present, there is no preparation and research of chimeric antigen receptor T cells targeting FGFR4

Method used

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  • FGFR4-targeting single-chain antibody, chimeric antigen receptor and chimeric antigen receptor T cell, preparation method of chimeric antigen receptor T cell, and application of FGFR4-targeting single-chain antibody, chimeric antigen receptor and chimeric antigen receptor T cell
  • FGFR4-targeting single-chain antibody, chimeric antigen receptor and chimeric antigen receptor T cell, preparation method of chimeric antigen receptor T cell, and application of FGFR4-targeting single-chain antibody, chimeric antigen receptor and chimeric antigen receptor T cell
  • FGFR4-targeting single-chain antibody, chimeric antigen receptor and chimeric antigen receptor T cell, preparation method of chimeric antigen receptor T cell, and application of FGFR4-targeting single-chain antibody, chimeric antigen receptor and chimeric antigen receptor T cell

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0079] A method for preparing a chimeric antigen receptor T cell targeting FGFR4, comprising the following steps:

[0080] (1) Provide the gene sequence of chimeric antigen receptor CAR-FGFR4 targeting FGFR4

[0081] Provide the nucleotide sequence (gene sequence of CAR-FGFR4) corresponding to any amino acid sequence shown in SEQ ID NO: 7-8, add the core corresponding to the amino acid sequence shown in SEQ ID NO: 9 at its 5' end Nucleotide sequence (gene sequence of signal peptide).

[0082] (2) Construction of pCDH-EF1-FGFR4-CAR recombinant plasmid

[0083] The above nucleotide sequence was inserted between the BamHI and EcoRI restriction sites of the pCDH-EF1-MCS vector, and behind the EF1α of the pCDH-EF1-MCS vector, EF1α was used as the promoter. When the above nucleotide sequence is inserted into the pCDH-EF1-MCS vector, a start codon (such as ATG) can be added to the 5' end of the above nucleotide sequence to connect with the BamHI restriction site in the pCDH-EF1-MCS...

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PUM

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Abstract

The invention provides a FGFR4-targeting single-chain antibody. Amino acid sequences of the FGFR4-targeting single-chain antibody comprises at least one kind of of the following sequences of (a) an amino acid sequence of a single-chain antibody heavy chain VH as shown in SEQ ID NO: 1 and an amino acid sequence of a single-chain antibody light chain VL as shown in SEQ ID NO: 2; and (b) an amino acid sequence of a single-chain antibody heavy chain VH as shown in SEQ ID NO: 3 and an amino acid sequence of a single-chain antibody light chain VL as shown in SEQ ID NO: 4. The invention also provides a chimeric antigen receptor and a chimeric antigen receptor T cell which comprise the FGFR4-targeting single-chain antibody. According to the chimeric antigen receptor and the chimeric antigen receptor T cell, FGFR4 can be specifically targeted, the in-vivo amplification of T cell is promoted, tumor cells are efficiently and specifically killed, the activity and lethality of cells are well maintained, and normal cells are not damaged.

Description

technical field [0001] The invention relates to the field of medical biology, in particular to a single-chain antibody targeting FGFR4, a chimeric antigen receptor, a chimeric antigen receptor T cell, and a preparation method and application thereof. Background technique [0002] Primary liver cancer is one of the most common malignant tumors and the second leading cause of cancer death worldwide, which has seriously threatened people's health and life. In clinical practice, the common treatment methods for liver cancer mainly include surgical resection, liver transplantation, chemotherapy and radiation therapy. However, since the early symptoms of liver cancer are not obvious and thus difficult to be detected, most patients are already in the middle and late stages when they are diagnosed, and lose the chance of surgical treatment and liver transplantation. In actual treatment, about 90% of patients with liver cancer need to receive drug treatment. Although the existing c...

Claims

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Application Information

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IPC IPC(8): C07K16/28C07K19/00C12N5/10C12N15/867A61K39/00A61P35/00
CPCC07K16/2863C07K14/7051C12N5/0636C12N15/86A61P35/00C07K2317/622C07K2319/03C07K2319/02C07K2319/33C12N2740/15043C12N2510/00A61K39/4611A61K39/464407A61K39/4631C07K16/28C07K19/00C12N5/10C12N15/867A61K39/395C12N15/63
Inventor 许晨光张宏玲温丽联唐超
Owner SHENZHEN BINDEBIOTECH CO LTD
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