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Chimeric antigen receptor with safety switch, expression gene of the chimeric antigen receptor, NK cell modified by the chimeric antigen receptor and an application of the chimeric antigen receptor.

A technology of chimeric antigen receptors and safety switches, which is applied in the field of NK cells and applications, chimeric antigen receptors and their expressed genes, can solve the problems of poor safety performance of NK cells, many side effects of treatment, and poor specificity

Active Publication Date: 2018-07-31
SHENZHEN MORECELL BIOMEDICAL TECH DEV CO LTD
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  • Abstract
  • Description
  • Claims
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AI Technical Summary

Problems solved by technology

[0005] However, traditional chimeric antigen receptor-modified NK cells have problems such as poor safety, many side effects, and poor specificity.

Method used

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  • Chimeric antigen receptor with safety switch, expression gene of the chimeric antigen receptor, NK cell modified by the chimeric antigen receptor and an application of the chimeric antigen receptor.
  • Chimeric antigen receptor with safety switch, expression gene of the chimeric antigen receptor, NK cell modified by the chimeric antigen receptor and an application of the chimeric antigen receptor.
  • Chimeric antigen receptor with safety switch, expression gene of the chimeric antigen receptor, NK cell modified by the chimeric antigen receptor and an application of the chimeric antigen receptor.

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Experimental program
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Effect test

Embodiment 1

[0057] 1. Construction of c-Met chimeric antigen receptor lentiviral vector with EGFRt switch

[0058] 1.1 c-Met scFv sequence

[0059] Using the anti-c-Met monoclonal antibody sequence screened by our laboratory, the VH and VL sequences were selected for the design of the scFv sequence in the CAR vector.

[0060] 1.2 Synthesis of c-Met CAR sequence with EGFRt switch

[0061] According to the sequence of each component of CAR, the full-length sequence contains signal peptide SP, c-Met single-chain antibody (scFv), CD8 hinge region and transmembrane domain, 41BB intracellular domain and DAP12-ITAM domain, T2A, Signal peptide SP, EGFRt, collectively referred to as SP-c-Met scFv-CD8-41BB-DAP12-T2A-SP-EGFRt, the structure diagram is as follows figure 1 shown. Restriction sites EcoR I and XhoI were added at the 5' and 3' ends of the entire framework, respectively, and Nanjing GenScript Biotechnology Co., Ltd. was entrusted to synthesize the entire expression cassette. The synthe...

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Abstract

The invention relates to a chimeric antigen receptor with a safety switch, an expression gene of the chimeric antigen receptor, an NK cell modified by the chimeric antigen receptor and an applicationof the chimeric antigen receptor. The chimeric antigen receptor comprises first signal peptide, c-Met single-chain antibody, CD8 structural domain, 41BB intracellular domain, DAP12-ITAM structural domain, T2A, second signal peptide and modified EGFRt. Experimental results show that the chimeric antigen receptor modified NK cells has features that the chimeric antigen receptor is controllable in immune response, the safety performance is good, the treatment side effect is less and the specificity is high.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a chimeric antigen receptor with a safety switch and its expression gene, its modified NK cell and its application. Background technique [0002] Human mesenchymal epithelial transition factor c-Met (Mesenchymal Epithelial Transition Factor, c-Met) is a receptor for hepatocyte growth factor (Hepatocyte Growth Factor, HGF), which belongs to the superfamily of tyrosine kinase receptors. By combining with HGF, c-Met activates the HGF / c-Met signaling pathway, reduces the adhesion between cells, and is beneficial to the metastasis of tumor cells. In recent years, studies have found that c-Met has abnormally high expression, mutation or activity changes in tumor tissues such as liver cancer, lung cancer, breast cancer, gastric cancer, pancreatic cancer, and glioma with a high incidence in my country. Therefore, c-Met has become an important target for antitumor therapy. [0003] Among th...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K19/00C12N15/62C12N5/10A61K35/17A61P35/00
CPCA61K35/17C07K14/7051C07K14/71C07K16/22C07K2319/02C12N5/0646
Inventor 不公告发明人
Owner SHENZHEN MORECELL BIOMEDICAL TECH DEV CO LTD
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