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Single-chain antibody targeting CD20, 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 that CART-CD19 can no longer exert anti-tumor effect and fail to kill tumor cells, achieve high infection efficiency and transcription efficiency, and promote Effects of amplified, persistent in vivo maintenance capacity

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

AI Technical Summary

Problems solved by technology

However, since tumors of the same type often express different immune phenotypes, for example, when tumor cells escape CD19-negative, that is, there are tumor cells that do not express CD19, the CD19 target will fail, and CART-CD19 will no longer be able to function. Anti-tumor effect, therefore, it is necessary to provide CAR-T cells that can target other broadly expressed targets of malignancies
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 CD20, chimeric antigen receptor T cell, preparation method and application thereof
  • Single-chain antibody targeting CD20, chimeric antigen receptor T cell, preparation method and application thereof
  • Single-chain antibody targeting CD20, chimeric antigen receptor T cell, preparation method and application thereof

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Embodiment 1

[0077] A method for preparing chimeric antigen receptor T cells targeting CD20, comprising the following steps:

[0078] (1) Prepare the gene sequence of chimeric antigen receptor CAR-CD20 targeting CD20

[0079] The coding gene sequences of signal peptide, single-chain antibody targeting CD20, CD8α hinge region, CD8 transmembrane region, 4-1BB signal region and CD3ζ signal region were respectively prepared, and the coding gene of the signal peptide is shown in SEQ ID NO: 15 The coding gene of the single-chain antibody targeting CD20 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 As shown in SEQ ID NO: 9, the gene encoding the 4-1BB signal region is shown in SEQ ID NO: 11, and the gene encoding the CD3ζ signal region is shown in SEQ ID NO: 13.

[0080] The coding gene sequences of the above-mentioned signal peptide, CD20-targeting single-chain antibody, CD8α hinge regio...

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PUM

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Abstract

The present invention provides a single-chain antibody targeting CD20, the single-chain antibody targeting CD20 comprises an amino acid sequence shown in SEQ ID NO: 1, and the single-chain antibody targeting CD20 is a humanized single-chain antibody. The present invention also provides a chimeric antigen receptor T cell comprising the single-chain antibody targeting CD20, wherein the chimeric antigen receptor targeting CD20 can specifically target CD20, especially can be targeted to tumor cells that do not express CD19, the expansion of T cells in patients is promoted, and the tumor cells canbe efficiently and specifically killed. In addition, the chimeric antigen receptor T cell targeting CD20 produced by humanized single-chain antibody has better cell viability and lethality. The invention also provides a preparation method and an application of the chimeric antigen receptor T cell targeting CD20.

Description

technical field [0001] The invention relates to the field of medical biology, in particular to a single-chain antibody targeting CD20, a chimeric antigen receptor T cell, and a preparation method and application thereof. Background technique [0002] 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. The treatment of Hodgkin's lymphoma has a significant effect and is considered to be one of the most effective treatments for malignant tumors. [0003] At present, CAR-T technology has achieved remarkable results in CD19-positive blood tumors (such as B-cell lymphoma, acute and chronic B-lymphoblastic leukemia). However, since tumors of the same type often express different immune phenotypes, for example, when tumor cells escape CD19-negative, that is, there are tumor cells that do not express CD19, the CD19 target will fail, and ...

Claims

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Application Information

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IPC IPC(8): C07K16/28C12N15/13C07K19/00C12N15/62C12N15/867C12N5/10A61P35/00
Inventor 曾滢张长风吴咏东
Owner SHENZHEN BINDEBIOTECH CO LTD
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