Preparation method of tumor neoantigen specific T cells

A specific and cell-based technology, applied in the field of cell biology, can solve the problems of low proportion of tumor neoantigen-specific T cells, limited killing effect of tumor cells, and low purity of mature DC cells, so as to achieve strong specific immune response and good stimulation Immune system, effect of improving phagocytosis efficiency

Pending Publication Date: 2022-01-18
HANGZHOU NEOANTIGEN THERAPEUTICS CO LTD
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Problems solved by technology

The immature DC cells obtained by this method have low purity due to containing T cells; directly lead to the low purity and small number of mature DC cells obtained by inducing immature DC cells; finally, tumor neogenesis obtained in the co-cultured cell suspension The proportion of antigen-specific T cells is extremely low, and the killing effect on tumor cells is very limited

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  • Preparation method of tumor neoantigen specific T cells
  • Preparation method of tumor neoantigen specific T cells
  • Preparation method of tumor neoantigen specific T cells

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preparation example Construction

[0053] A method for preparing tumor neoantigen-specific T cells and verifying their effectiveness in killing tumor cells, comprising:

[0054] Step 1, synthesizing tumor neoantigen polypeptide;

[0055] For specific methods, please refer to a screening method for individualized tumor neoantigen peptides in patent 202110251383.X to obtain polypeptide sequences, and then synthesize polypeptides, and use solid-phase synthesis technology to produce suitable purity polypeptides; to verify the immune function of the obtained polypeptides:

[0056] Blood samples from healthy volunteers were drawn to separate PBMCs, and the immune function of the peptide vaccine was verified in vitro, and the secretion of IFN-γ after peptide stimulation was detected. Using the ELISpot method, with the polypeptide vaccine as the stimulus, the long polypeptide of the present invention as the experimental group 1, the corresponding short polypeptide at the same site in the long polypeptide as the experim...

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Abstract

The invention discloses a preparation method of tumor neoantigen specific T cells, which comprises the following steps: step 1, obtaining biological information through gene sequencing, and designing and synthesizing tumor neoantigen polypeptide; step 2, loading the tumor neoantigen polypeptide to immature DC cells, and inducing the DC cells to mature by using cytokines; step 3, performing multi-round co-incubation stimulation on the mature DC cells and the initial T cells; step 4, separating and collecting tumor neoantigen specific T cells by using an immunomagnetic bead technology; step 5, amplifying to obtain tumor neoantigen specific T cells with high purity and killing activity; step 6, performing cryopreservation to obtain tumor neoantigen specific T cells; according to the method disclosed by the invention, a large number of tumor neoantigen specific T cells with high purity and high killing activity can be efficiently prepared, and the tumor killing efficiency is close to 100%; the cost is low, the speed is high, and market requirements are met.

Description

technical field [0001] The invention relates to the technical field of cell biology, in particular to a method for preparing tumor neoantigen-specific T cells. Background technique [0002] Recently, due to breakthroughs in the treatment of malignant melanoma and blood system tumors, adoptive immune cell therapy has once again become a hot spot in the field of tumor immunotherapy research. At present, the commonly used effector cells of adoptive immune cell therapy in the world mainly include tumor infiltrating lymphocytes (Tumor Infiltrating Lymphocyte, abbreviated as TIL), cytokine-induced killer cells (Dendritic Cell-Cytokine Induced Killer, abbreviated as DC-CIK), chimeric Antigen receptor modified T cells (Chimeric Antigen Receptor T-Cell, abbreviated as CAR-T) and T cell receptor genetically engineered T cells (T-Cell Receptor gene engineered T-cell, abbreviated as TCR-T). For adoptive immune cell therapy, the acquisition of highly specific tumor antigen targets is ex...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0783C12N5/0784
CPCC12N5/0636C12N2502/1121C12N2501/599C12N2501/25C12N2501/2301C12N2501/2306C12N2501/02C12N2501/999C12N2501/515C12N2501/51C12N2501/52C12N2501/2302C12N2501/2307C12N2501/2315
Inventor 莫凡刘亮丁冬
Owner HANGZHOU NEOANTIGEN THERAPEUTICS CO LTD
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