Mode cell for expressing anti-human DSG3 scFv on cell surface and construction method thereof

A cell and model technology, applied in the field of molecular biology, can solve problems such as high price, high recurrence rate, and heavy burden

Pending Publication Date: 2022-06-28
中国人民解放军西部战区总医院
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Since all clones of B cells in the immune system express CD20 molecules, there are clinical studies in the United States using the monoclonal antibody drug Rituximab (trade name: MabThera) developed by Roche in recent years to target CD20 to eliminate B cells. Herpes, although it can re...

Method used

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  • Mode cell for expressing anti-human DSG3 scFv on cell surface and construction method thereof
  • Mode cell for expressing anti-human DSG3 scFv on cell surface and construction method thereof
  • Mode cell for expressing anti-human DSG3 scFv on cell surface and construction method thereof

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Embodiment 1

[0047] By artificially synthesizing a DNA fragment containing the nucleotides shown in SEQ ID No.6, the specific sequence of the DNA fragment is shown in SEQ ID No.7, and the structure is shown in figure 2 shown. The fragment includes CD8-leader leader region, scFv region (scFv fragment of humanized DSG3-specific single chain antibody), CD8-hinge hinge region and CD8-TM region (transmembrane region). The specific sequence of SEQ ID No.7 is as follows:

[0048] GTCGACGCCACCATGGCCCTGCCCGTGACCGCCCTGCTGCTGCCCCTGGCCCTGCTGCTGCACGCCGCCAGGCCCCAGGTGCAGCTGGTGCAGAGCGGCGCCGAGCTGGTGAGGCCCGGCGCCAGCGTGAAGCTGAGCTGCCAGGCCAGCGGCTACACCTTCACCAGCCACTACATCCACTGGGTGAGGCAGGCCCCCGGCCAGGAGAGGGAGTGGGGCTGGATCAACCCCAGCGGCGGCAAGACCAACACCAACCAGAACTTCAAGGACAAGGCCACCCTGACCGTGGACAAGAGCAGCAGCGCCGCCTACACCAGCGAGGACAGCGCCGTGTACTTCTGCGCCAGGGACCAGAGCCTGGGCATGGACGTGTGGGGCCAGGGCACCCTGGTGACCAGCGCCGGCGGCGGCGGCAGCGGCGGCGGCGGCAGCGACATCCAGATGACCCAGAGCCCCAGCAGCCTGAGCGCCAGCGTGGGCGACAGGGTGACCATCACCTGCAGCAGCGACATCGGCAGGTACAAC...

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Abstract

The invention discloses a model cell capable of expressing anti-human DSG3scFv on the cell surface and a construction method of the model cell, an artificially designed membrane type anti-DSG3 single-chain antibody is expressed on the surface of the model cell, the model cell can be combined with a T cell expressing an anti-DSG3 autoantibody receptor (CAAR), namely, a CAAR-T cell, the model cell is used for identifying a killing evaluation system of the chimeric autoantibody receptor (CAAR) CAAR-T cell, and the model cell can be used for identifying the killing evaluation system of the CAAR-T cell. And a simple, convenient and easily available experimental tool is provided for the application of the CAAR-T cells.

Description

technical field [0001] The invention belongs to the field of molecular biology, in particular to a model cell expressing anti-human DSG3 scFv on the cell surface and a construction method thereof. Background technique [0002] In recent years, chimeric antigen receptor (CAR)-T cells have brought remarkable innovative progress and efficacy to tumor therapy. Taking advantage of the fact that CD8+ T cells can differentiate into CTL cells that can effectively kill target cells after activation, gene modification technology is used to express an artificially designed transmembrane protein molecule on the surface of CD8+ T cells: the extracellular region is a specific recognition antigen Molecular single-chain antibody (single chain fragment variable, SCFV), also known as antigen receptor (antigen receptor), the intracellular region is TCR / CD3ζ and costimulatory molecule intracellular region part. This artificial chimeric antigen receptor molecule is commonly referred to as a CAR...

Claims

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Application Information

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IPC IPC(8): C12N5/10C07K19/00C12N15/62C12N15/867
CPCC12N5/0694C07K16/18C07K14/70517C12N15/86C12N2510/00C07K2317/622C07K2317/24C07K2319/03C07K2319/33C12N2740/15043C12N2800/107
Inventor 李华刘佳慧毛娟
Owner 中国人民解放军西部战区总医院
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