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Multi-signal chimeric antigen receptor and its expression gene, its modified nk cell and application

A chimeric antigen receptor and gene expression technology is applied to genetically modified cells, cells modified by introducing foreign genetic material, animal cells, etc. It can solve the problems of insensitive NK cell recognition process and many side effects of treatment

Active Publication Date: 2020-07-14
SHENZHEN MORECELL BIOMEDICAL TECH DEV CO LTD
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] However, the traditional NK cells modified by chimeric antigen receptors have the problems of insensitive recognition process and many side effects of treatment

Method used

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  • Multi-signal chimeric antigen receptor and its expression gene, its modified nk cell and application
  • Multi-signal chimeric antigen receptor and its expression gene, its modified nk cell and application
  • Multi-signal chimeric antigen receptor and its expression gene, its modified nk cell and application

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Embodiment 1

[0058] 1. Construction of lentiviral expression vector

[0059] 1.1c-Met scFv sequence

[0060] 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.

[0061] 1.2 Sequence synthesis and vector construction

[0062] According to the experimental requirements, three sequences were synthesized for the construction of lentiviral expression vectors:

[0063] Expression vector one (group one): pLenti7.3-γδTCR lentiviral expression vector, the sequence of which is shown in SEQ ID No.3, including signal peptide SP, TCRγ chain, and TCRδ chain.

[0064] Expression vector two (group two): pLenti7.3-c-Met-CAR lentiviral expression vector, the sequence is shown in SEQID No.4, including c-Met single-chain antibody (scFv), CD8 hinge region and transmembrane structure domain, 41BB intracellular domain and DAP12-ITAM domain.

[0065] Expression vector three (the third group)...

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Abstract

The invention relates to a multi-signal chimeric antigen receptor and its expression gene, its modified NK cell and its application. The chimeric antigen receptor includes a first fragment, a second fragment, and a self-cleaving polypeptide T2A connecting the first fragment and the second fragment. The first fragment includes a first signal peptide, a TCRγ chain and a TCRδ chain sequentially connected, and the second fragment Including the second signal peptide SP, tumor antibody, CD8 domain, 41BB intracellular domain and DAP12-ITAM domain connected in sequence, the C-terminus of the self-cleaving polypeptide T2A is connected to the TCRδ chain, and the N-terminus of the self-cleaving polypeptide T2A is connected to the first The two signal peptides SP are connected. Experimental results show that the above-mentioned multi-signal chimeric antigen receptor can be stably expressed in NK cells, can more quickly and specifically recognize and kill malignant tumor cells, and reduce the toxic and side effects of treatment.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a multi-signal chimeric antigen receptor and its expression gene, its modified NK cell and its application. Background technique [0002] Metabolism is the basic feature of the body's life activities, including material metabolism and accompanying energy metabolism. The release, conversion and utilization of energy in the process of material metabolism of organisms is called energy metabolism. An important feature of malignant tumors is the high metabolic capacity of tumor cells, which can specifically overproduce small tumor phosphorylated antigen molecules. [0003] In recent years, adoptive cell therapy technology, chimeric antigen receptor (chimeric antigen receptor, CAR) modified T cell technology has made a significant breakthrough in the clinical application of tumor treatment, showing good targeting in clinical trials , lethality and persistence, showing great application p...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K19/00C12N15/62C12N15/867C12N5/10A61K35/17A61P35/00
CPCC12N5/0646C12N15/86C07K14/7051C07K16/30C12N2510/00C12N2740/15043C12N2800/107C07K2319/33C07K2319/03C07K2319/00C07K2317/622C07K2319/74A61K39/46441A61K2239/53A61K39/4613A61K39/4631C07K16/2863C07K2317/73A61K2039/844A61P35/00C12N2740/16043C07K14/70517
Inventor 不公告发明人
Owner SHENZHEN MORECELL BIOMEDICAL TECH DEV CO LTD
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