Multi-signal embedded antigen receptor and expression gene thereof, NK (Natural Killer) cell modified by multi-signal embedded antigen receptor and application

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

Active Publication Date: 2018-07-24
SHENZHEN MORECELL BIOMEDICAL TECH DEV CO LTD
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  • Summary
  • 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 embedded antigen receptor and expression gene thereof, NK (Natural Killer) cell modified by multi-signal embedded antigen receptor and application
  • Multi-signal embedded antigen receptor and expression gene thereof, NK (Natural Killer) cell modified by multi-signal embedded antigen receptor and application
  • Multi-signal embedded antigen receptor and expression gene thereof, NK (Natural Killer) cell modified by multi-signal embedded antigen receptor and application

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Experimental program
Comparison scheme
Effect test

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 embedded antigen receptor and an expression gene thereof, an NK (Natural Killer) cell modified by the multi-signal embedded antigen receptor and application. The embedded antigen receptor comprises a first segment, a second segment and a self-pyrolysis polypeptide T2A for connecting the first segment and the second segment, wherein the first segment comprises a first signal peptide, a TCRgamma chain and a TCRdelta chain, which are connected in sequence; the second segment comprises a second signal peptide SP, a tumor antibody, a CD8 structure domain, a 41BB intracellular domain and a DAP12-ITAM structural domain, which are connected in sequence; a C end of the self-pyrolysis polypeptide T2A is connected with the TCRdelta chain; an N end of the self-pyrolysis polypeptide T2A is connected with the second signal peptide SP. An experiment result shows that the multi-signal embedded antigen receptor can be stably expressed in the NK cell and can be used for more rapidly and specifically identifying malignant tumor cells and killing the malignant tumor cells, and the toxic side effect of treatment is reduced.

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 Applications(China)
IPC IPC(8): C07K19/00C12N15/62C12N15/867C12N5/10A61K35/17A61P35/00
CPCA61K35/17C12N5/0646C12N15/86C07K14/7051C07K16/30C12N2510/00C12N2740/15043C12N2800/107C07K2319/33C07K2319/03C07K2319/00C07K2317/622C07K2319/74C07K16/2863C07K2317/73A61K2039/5156A61K2039/844A61K39/00111A61K39/001129A61P35/00C12N2740/16043C07K14/70517
Inventor 不公告发明人
Owner SHENZHEN MORECELL BIOMEDICAL TECH DEV CO LTD
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