Construction method of LRFFT1 cells

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

Active Publication Date: 2019-08-06
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 effective analysis of patient antigens

Method used

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  • Construction method of LRFFT1 cells
  • Construction method of LRFFT1 cells
  • Construction method of LRFFT1 cells

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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 present invention discloses a construction method of LRFFT1 cells. The construction method of the LRFFT1 cells comprises the following steps: ctDNA sequencing of human peripheral blood or whole exon sequencing of tumor tissues is conducted to screen out mutation sites for antigen epitope prediction, and a mutant polypeptide expression gene sequence is ligated and synthesized; simultaneously alentiviral vector is constructed, lentiviruses are packaged, the packaged lentiviruses transfect APC cells, modification of specific LV cells is completed, the APC cells are co-cultured with PBMC isolated from peripheral blood in vitro, effective peptides are screened out, and ordinary T cells are transformed into RFF cells with a more precise killing ability by a second impact of a precise and effective peptide stimulation; then a TCR-T technology principle is used to conduct modification; and the modified T cells are then subjected to a gene-editing technology to conduct knocking out of immunosuppressive targets, the T cells with the specific killing ability is precisely protected from in vivo inhibition, the killing ability of the T cells on tumor cells is improved, and the LRFFT1 cellsmore combining attack and defense are 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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Patent Type & Authority Applications(China)
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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