A kind of construction method of afft2 cell

Abstract

The invention belongs to the field of biotechnology, and in particular relates to a method for constructing AFFT2 cells. This method transforms cells with TCR-T technology, and the transformed T cells are blocked in vitro with inhibitory signal molecule antibody drugs, thereby effectively improving the anti-tumor ability of T cells. Cells transformed by the AFFT2 program can recognize tumor antigens The proportion of specific T cells is above 70%.

Description

Technical field:

[0001] The invention belongs to the field of biotechnology, and in particular relates to an AFFT2 cell and a preparation method thereof. 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 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.

[0003] 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.

[0004] The above treatment methods are immature, especially the in vitro induction of DC ...

Examples

Embodiment 1

[0051] This example will take lung cancer patients as an example to provide targeted AFFT2 cells and their preparation methods:

[0052] 1. Whole Exome Sequencing

[0053] 1) Take peripheral blood from lung cancer patients for ctDNA sequencing and HLA typing;

[0054] 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;

[0055] 2. Epitope prediction

[0056] 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";

[0057] 2) Use prediction software to analyze the IC50 of the potential antigenic epitope (recommended software: NetMHCpan 3.0, PickPocket, artificial neural networks (ANN)), if IC50<1000nM, the potential antigenic epitope is considered as "antigenic epitope";

[0058] 3. Synthetic peptides

[0059] Entrust a technical service company to synthesize a mut...

Technical field:

[0001] The invention belongs to the field of biotechnology, and in particular relates to an AFFT2 cell and a preparation method thereof. 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 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.

[0003] 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.

[0004] The above treatment methods are immature, especially the in vitro induction of DC ...

Examples

Embodiment 1

[0051] This example will take lung cancer patients as an example to provide targeted AFFT2 cells and their preparation methods:

[0052] 1. Whole Exome Sequencing

[0053] 1) Take peripheral blood from lung cancer patients for ctDNA sequencing and HLA typing;

[0054] 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;

[0055] 2. Epitope prediction

[0056] 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";

[0057] 2) Use prediction software to analyze the IC50 of the potential antigenic epitope (recommended software: NetMHCpan 3.0, PickPocket, artificial neural networks (ANN)), if IC50<1000nM, the potential antigenic epitope is considered as "antigenic epitope";

[0058] 3. Synthetic peptides

[0059] Entrust a technical service company to synthesize a mut...

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