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Single-chain antibody targeting CD317, 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, can solve the problems of failure to kill tumor cells, limitations, and no progress, achieve high infection efficiency and transcription efficiency, promote amplification, and persist. The effect of maintaining capacity in the body

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

However, the current application of CAR-T technology is still limited to hematological tumors, and has not yet made progress in various solid tumors such as liver cancer.
In addition, the in vivo maintenance ability (persistence) of CAR-T cells is the guarantee of the long-term efficacy of CAR-T, but in fact, CAR-T completely disappears after a few days to a few weeks after reinfusion, and fails to exert a long-lasting and lasting effect. ability to kill tumor cells

Method used

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  • Single-chain antibody targeting CD317, chimeric antigen receptor T cell, preparation method and application thereof
  • Single-chain antibody targeting CD317, chimeric antigen receptor T cell, preparation method and application thereof
  • Single-chain antibody targeting CD317, chimeric antigen receptor T cell, preparation method and application thereof

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Experimental program
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Embodiment 1

[0077] A method for preparing chimeric antigen receptor T cells targeting CD317, comprising the following steps:

[0078] (1) Prepare the gene sequence of chimeric antigen receptor CAR-CD317 targeting CD317

[0079]The coding genes for the signal peptide, the single-chain antibody targeting CD317, the CD8α hinge region, the CD8 transmembrane region, the 4-1BB signal region and the CD3ζ signal region were respectively prepared, and the coding gene sequence of the signal peptide is shown in SEQ ID NO: 15 The coding gene sequence of the single-chain antibody targeting CD317 is shown in SEQ ID NO: 2, the coding gene sequence of the CD8α hinge region is shown in SEQ ID NO: 7, and the coding gene sequence of the CD8 transmembrane region The sequence is shown in SEQ ID NO: 9, the coding gene sequence 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.

[0080] The coding gene sequences of the above-mentioned ...

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PUM

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Abstract

The present invention provides a single-chain antibody targeting CD317, the single-chain antibody targeting CD317 comprises an amino acid sequence shown in SEQ ID NO: 1, and the single-chain antibodytargeting CD317 is a humanized single-chain antibody. The present invention also provides a chimeric antigen receptor T cell comprising the single-chain antibody targeting CD317, wherein the chimericantigen receptor targeting CD317 can specifically target CD317, the expansion of T cells in patients is promoted, the tumor cells can be efficiently and specifically killed, and damage is hardly generated on the normal cells. In addition, the chimeric antigen receptor T cell targeting CD317 produced by humanized single-chain antibody can keep the ability of self-renewal and tumour lethality. The invention also provides a preparation method and an application of the chimeric antigen receptor T cell targeting CD317.

Description

technical field [0001] The invention relates to the field of medical biology, in particular to a single-chain antibody targeting CD317, a chimeric antigen receptor T cell, and a preparation method and application thereof. Background technique [0002] Malignant tumor (cancer) has become the culprit threatening human life, and the incidence rate is on the rise. CD317 is a lipid raft-related protein, which is mainly expressed in various solid tumor cells such as liver cancer and colon cancer, but rarely expressed in ordinary cells. The expression of CD317 is often the response of cells to the gamma-interferon pathway, so it is closely related to the occurrence of tumors. [0003] CAR-T (Chimeric Antigen Receptor T Cell) technology is a new type of cell therapy, which is to reinfuse CAR-modified T cells into the human body, activate the autoimmune system, and kill tumor cells. One of the most effective treatments for malignant tumors. However, the current application of CAR-...

Claims

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Application Information

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IPC IPC(8): C07K16/28C12N15/13C07K19/00C12N15/62C12N15/867C12N5/10A61P35/00
Inventor 张宏玲龙丽梅
Owner SHENZHEN BINDEBIOTECH CO LTD
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