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Construction method of AFF cells

A construction method and cell technology, applied in animal cells, vertebrate cells, cell culture active agents, etc., can solve problems such as T cell self-protection technology, and achieve strong target specificity, high ratio, and safety high effect

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

AI Technical Summary

Problems solved by technology

[0007] None of the above schemes consider the self-protection technology of T cells, so that a small number of specific T cells directly face the powerful tumor microenvironment

Method used

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  • Construction method of AFF cells
  • Construction method of AFF cells
  • Construction method of AFF cells

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] This example will take lung cancer patients as an example to provide targeted AFF cells and their preparation methods:

[0038] 1. Whole Exome Sequencing

[0039] 1) Take peripheral blood from lung cancer patients for ctDNA sequencing and HLA typing;

[0040] 2) Use software to analyze the sequencing information: compare the ctDNA sequencing results with the genome of normal cells, and screen out the mutation sites;

[0041] 2. Epitope prediction

[0042] 1) Take the mutated amino acid site as the center, extend 10 amino acids to both sides, and use this 21-amino acid polypeptide as a "potential antigenic epitope";

[0043] 2) Use prediction software to analyze the IC50 of potential antigenic epitopes (recommended software: NetMHCpan 3.0, PickPocket, artificial neural networks (ANN)), if IC50<1000nM, consider this potential antigenic epitope as "antigenic epitope";

[0044] 3. Synthetic peptides

[0045] Entrust a technical service company to synthesize a mutant pol...

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Abstract

The invention belongs to the technical field of biology, and particularly relates to a construction method of AFF cells. The construction method of the AFF cells comprises the following main steps: 1)drawing peripheral blood of a patient, performing ctDNA exon sequencing, or performing full exon sequencing on tumor tissues, screening out mutation sites, performing epitope prediction, and synthesizing mutant polypeptides; and 2) preparing immortalized DC by using the peripheral blood, loading the immortalized DC with the mutant polypeptides, and co-incubating the immortalized DC with PBMC to obtain the AFF cells.

Description

[0001] Technical field: [0002] The invention belongs to the field of biotechnology, and in particular relates to a method for constructing AFF cells. [0003] Background technique: [0004] At present, in terms of tumor-specific immunotherapy, the existing LAK, DC, CIK, and DC-CIK cells and methods have basically been proven to be ineffective, while NK, CAR-NK, TIL, and other cell technologies have yet to mature. CAR - T cells have shortcomings in safety and solid tumor therapy. [0005] 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. [0006] The above treatment methods are immature, especially the in vitro induction of...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0783C12N5/0784
CPCC12N5/0636C12N2501/998C12N2502/1121
Inventor 焦顺昌张嵘周子珊陈小彬张天赋王海燕陈红利
Owner BEIJING DCTY BIOTECH CO LTD
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