LRFFT1 cell construction method

A construction method and cell technology, applied in the field of LRFFT1 cell construction, can solve the problems of lack of patient antigens, lack of accurate and effective analysis, etc., and achieve the effect of strong target specificity, less off-target effect, and improved lethality

Inactive Publication Date: 2019-01-04
BEIJING DCTY BIOTECH CO LTD
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

There is a general lack of accurate and

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  • LRFFT1 cell construction method
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  • LRFFT1 cell construction method

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[0060] The present invention is described below through specific embodiments. Unless otherwise specified, the technical means used in the present invention are all methods known to those skilled in the art. In addition, the embodiments should be understood as illustrative rather than limiting the scope of the present invention, and the spirit and scope of the present invention are only defined by the claims. For those skilled in the art, without departing from the essence and scope of the present invention, various changes or modifications to the material components and dosages in these embodiments also belong to the protection scope of the present invention.

[0061] The technical solution is detailed as follows:

[0062] 1. Antigenic epitope prediction

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

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

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Abstract

The invention provides an LRFFT1 cell construction method, human peripheral blood is used to sequence ctDNA or a whole exome of a tumor tissue, a mutation site is screened to predicate an epitope, andthe sequence of an expression gene of a mutation polypeptide is connected and synthesized; meanwhile, a lentiviral vector is constructed, a lentivirus is packaged, an APC (Antigen Presenting Cell) istransfected to finish the transformation of a specific LV cell, the APC is cultured in vitro together with a PBMC (Peripheral Blood Mononuclear Cell) separated from the peripheral blood, the effective polypeptide is screened, a common T cell is transformed into an RFF cell having more accurate lethality through the second impact stimulated by the accurate effective polypeptide; the transformationis conducted through the TCR-T technical principle; an immunosuppressive target of the transformed T cell is then knocked off through the gene editing technology, the specific lethal T cell is protected accurately against in-vivo suppression, the lethality of the T cell on a tumor cell is improved, and a more offensive and defensive LRFFT1 cell is obtained.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a method for constructing LRFFT1 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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IPC IPC(8): C12N15/90C12N5/10
CPCC07K14/7051C12N5/0638C12N15/907C12N2510/00C12N2800/80C12N2810/10
Inventor 焦顺昌张嵘张天赋周子珊宋玉洁陈小彬吴子明
Owner BEIJING DCTY BIOTECH CO LTD
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