Method for constructing MRFFT1 cell

A construction method and cell technology, which is applied in the field of MRFFT1 cell construction, can solve the problem of not considering the self-protection technology of T cells, and achieve the effects of strong target specificity, increased lethality, and less off-target effects

Active Publication Date: 2019-08-06
BEIJING DCTY BIOTECH CO LTD +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] None of the above schemes considers the self-protection technology of T cells, so that a small number of specific T cells directly face the powerful tumor immune microenvironment

Method used

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  • Method for constructing MRFFT1 cell
  • Method for constructing MRFFT1 cell
  • Method for constructing MRFFT1 cell

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Embodiment Construction

[0060] The present invention is described below through specific embodiments. Unless otherwise specified, the technical means used in the present invention are methods known to those skilled in the art. In addition, the embodiments should be considered as illustrative rather than limiting the scope of the invention, the spirit and scope of which is defined only by the claims. For those skilled in the art, on the premise of not departing from the spirit and scope of the present invention, various changes or modifications to the material components and dosage in these embodiments also belong to the protection scope of the present invention.

[0061] The technical solution is detailed as follows:

[0062] 1. Epitope prediction

[0063] 1) Use the peripheral blood of lung cancer patients for ctDNA sequencing and HLA typing detection;

[0064] 2) Use software to analyze the sequencing information: compare the ctDNA sequencing results with the genome of normal cells, and screen o...

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Abstract

The invention relates to a method for constructing MRFFT1 cells. The method comprises the following steps of: screening out a mutant polypeptide through antigen epitope prediction, connecting and synthesizing a mutant polypeptide expression gene sequence; and meanwhile, constructing an MVA virus vector, packaging MVA virus, transfecting APC cells, completing the modification of specific MV cells,co-culturing PBMC separated from peripheral blood with the modified specific MV cells in vitro, screening out effective polypeptide, modifying common T cells into RFF cells with more accurate killingcapacity through the second impact of accurate effective polypeptide stimulation, performing modification with a TCR-T technology principle, and knocking out immunosuppressive target of the modified Tcells by using a gene editing technology. The method accurately protects specific killing T cells from in-vivo inhibition, and improves the killing ability of T cells to tumor cells.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a method for constructing MRFFT1 cells. Background technique [0002] At present, in terms of tumor-specific immunotherapy, the existing LAK, DC, CIK, and DC-CIK cells and methods have basically been proved to be ineffective, while NK, CAR-NK, TIL and other cell technologies have yet to mature. CAR- T cells also have shortcomings in safety and solid tumor therapy. In the prior art, DCs are generally modified to present T cells to produce specific killing. Some laboratories are trying to use viruses as vectors to transfect and present T cells to induce specific killing of T cells. We have also used mutant hybrid peptides to directly stimulate PBMCs to induce T cells. There are also laboratories using TCR-T technology to target the presentation of MAGE A3 antigen. [0003] The above treatment methods are immature, especially the in vitro induction of DC cells and the te...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/90C12N5/10
CPCC07K14/7051C12N5/0638C12N15/907C12N2510/00C12N2800/80C12N2810/10
Inventor 焦顺昌张嵘周子珊宋玉洁解佳森张天赋王海燕袁翰
Owner BEIJING DCTY BIOTECH CO LTD
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