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A method for testing the immunogenicity of neoantigens

A technology of antigen immunization and testing methods, which is applied in the fields of cell biology and bioinformatics, can solve technical difficulties and other problems, and achieve the effects of simplifying the experimental process, improving work efficiency, and ensuring accuracy

Active Publication Date: 2022-04-05
深圳裕泰抗原科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

This method is technically difficult in actual operation and is rarely used.

Method used

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  • A method for testing the immunogenicity of neoantigens
  • A method for testing the immunogenicity of neoantigens
  • A method for testing the immunogenicity of neoantigens

Examples

Experimental program
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Embodiment 1

[0080] This embodiment provides a method for testing the immunogenicity of neoantigens. The immunogenicity determination process for the above five antigenic polypeptides #1-#5 is as follows:

[0081] S1, from 100ml of peripheral blood, separated by ficoll, 1.2*10 8 PBMCs.

[0082] S2. Sorting by CD8 magnetic beads to obtain 2.4*10 from PBMC 7 CD8 positive T cells, with 10% FBS+90% DMSO according to 5*10 6 / ml density to freeze CD8 positive T cells. When the DC cells are mature, they can be resuscitated and used again.

[0083] S3. After the CD8-positive T cells are sorted, the remaining CD8-negative T cells are obtained. Spread the CD8-negative T cells in a 6-well plate, and the monocytes will adhere to the wall, and the suspension cells are removed.

[0084] S4, culturing the DC cells, adding cytokines to the adherent and isolated monocytes to induce DC cells, and obtaining mature DC cells after culturing for 7 days, and collecting the mature DC cells.

[0085] S5. The ...

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Abstract

The invention discloses a method for testing the immunogenicity of neoantigens. By dividing multiple antigenic polypeptides into several groups and using the method of separate loading and co-stimulation, the operational complexity of the experiment is significantly reduced, and the experimental process is greatly simplified. At the same time, the accuracy of the experimental results is guaranteed. Compared with the conventional single-load and single-stimulation method, T cells can be stimulated by multiple polypeptides. When testing, it is only necessary to detect the specificity of different neoantigen polypeptides contained in one T cell. Sexual T cells, the testing process has been significantly simplified, improving work efficiency and reducing testing costs.

Description

technical field [0001] The invention belongs to the technical field of cell biology and bioinformatics, and relates to an immunogenicity detection method, in particular to a neoantigen immunogenicity detection method. Background technique [0002] Usually, the method to verify the immunogenicity of neoantigens is to isolate peripheral blood mononuclear cells (PBMC) from the peripheral blood of tumor patients or healthy volunteers, and to sort out T cells from PBMC, or to isolate tumor cells from tumor tissues. Infiltrating lymphocytes (TIL) and sorting out T cells; thus, the immunogenicity of neoantigens was verified by enzyme-linked immunospot method (Elispot) for peripheral blood-derived T cells or TIL-derived T cells. For T cells derived from peripheral blood, since the frequency of neoantigen-specific T cells contained therein is very low, even lower than the detection limit of the most sensitive experimental method, it is usually necessary to use pre-stimulation first, ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/0783C12N5/0784G01N33/68
CPCC12N5/0636C12N5/0639G01N33/6854G01N33/6866C12N2502/00C12N2501/2302C12N2501/2307C12N2501/2315G01N2333/57
Inventor 唐云霞王煜高志博李淼张义兴朱小娟
Owner 深圳裕泰抗原科技有限公司
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