Method of preparing CAR-T cell from CD3

A cell-positive technology, applied in the field of molecular biology, can solve the problems of epithelial malignant tumors that need to be improved, and achieve the effect of easy industrial production, simple preparation method and high safety

Inactive Publication Date: 2018-11-02
沈阳美达博生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the safety and effectiveness of CAR-T cells in the treatment of epithelial malignancies need to be improved

Method used

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Embodiment Construction

[0030] In order to make the object, technical solution and advantages of the present invention more clear, the present invention will be further described in detail below in conjunction with examples, but the examples given are not intended to limit the present invention.

[0031] In the embodiment of the present invention, the CAR-T cells are obtained by infecting CD3-positive T cells with a virus containing CAR.

[0032] CD3 positive T cells are prepared by the following steps:

[0033] S10, Isolation of T cells

[0034] S101. Draw 20ml of blood into an anticoagulant tube, or add 0.1ml (125-250) / ml heparin solution to every 1ml of whole blood, shake well, put them together in a 50ml centrifuge tube, add 20ml of PBS (equal to whole blood) to dilute;

[0035] S102. Add 3ml of lymphatic separation solution to the 15ml centrifuge tube in advance;

[0036] S103. Use a graduated pipette along the inclined tube wall, slowly superimpose the diluent and the layered liquid surface, ...

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Abstract

The invention provides a method of preparing a CAR-T (chimeric antigen receptor T) cell from CD3, and relates to the technical field of molecular biology. The method comprises the steps of infecting aCD3 positive T cell with a virus containing a CAR to form the CAR-T cell. The prepared CAR-T cell has high effectiveness and safety, and the method is simple, practicable, and easy in industrial production.

Description

technical field [0001] The invention relates to the technical field of molecular biology, in particular to a method for preparing CAR-T cells by using CD3. Background technique [0002] With the development of genetic engineering technology and the improvement of understanding of T cell recognition, tumor targeting receptor-CAR came into being, so there is also the possibility of transfecting CAR into human T cells to enhance the antigen recognition specificity of adoptive immunotherapy and lethal technology. The design basis of CAR is to link the extracellular ligand recognition region—typically the single-chain variable region—scFv fragment with intracellular signaling molecules including CD3ζ, and then effectively activate T cells by interacting with antigens . CAR-T cells designed based on TCR signaling have been successfully investigated in the treatment of refractory B-cell malignancies. However, the safety and effectiveness of CAR-T cells in the treatment of epithe...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/10
CPCC12N5/0636C12N2501/515C12N2510/00
Inventor 隋长江
Owner 沈阳美达博生物科技有限公司
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