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Method for screening tumor neoantigen based on HLA typing and structure

An antigen and tumor technology, applied in the field of tumor neoantigen screening, can solve the problems of high cost, time-consuming, labor-intensive, etc., and achieve the effect of saving labor and funds, reducing the number of experiments, and saving funds.

Active Publication Date: 2020-01-10
BETA PHARM SUZHOU LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] However, the development of tumor vaccines is currently screened one by one by means of experiments. This process is time-consuming, laborious, and expensive, and it is difficult to find suitable tumor vaccines (tumor antigens). The method of screening tumor neoantigens is to obtain a suitable tumor vaccine by screening tumor neoantigens, so as to facilitate subsequent further targeted experiments based on this, so as to greatly reduce the number of experiments, and achieve time-saving, labor-saving and cost-saving

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  • Method for screening tumor neoantigen based on HLA typing and structure
  • Method for screening tumor neoantigen based on HLA typing and structure
  • Method for screening tumor neoantigen based on HLA typing and structure

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Embodiment 1

[0050] Such as figure 1 As stated, this embodiment provides a method for screening tumor neoantigens based on HLA typing and structure, including the steps of:

[0051] S101. Obtain the polypeptide sequence encoded by the mutant gene of the patient's tumor tissue cells. Specifically, including:

[0052] A. Extract the DNA of tumor tissue cells by SDS method, and perform DNA sequencing on it;

[0053] B. Comparing the sequenced DNA sequence with the normal wild-type DNA sequence of the tissue cell to obtain a mutated DNA sequence different from the normal wild-type DNA sequence of the tissue cell. Wherein, the normal DNA sequence of the tissue cells may be acquired through an existing database. Wherein, the database may be: COSMIC, NCBI, UCSC, Ensembl, TCGA, etc.

[0054] C. Obtain the polypeptide sequence encoded by the mutated DNA sequence through biological software. Wherein, the biological software can be DNA-man, or other software that can translate DNA sequences into...

Embodiment 4

[0115] Example 4: Evaluation of Neoantigenic Peptide Activity:

[0116] 4.1 T2 cell culture: T2 cells were purchased from ATCC and cultured with 20% FBS IMDM (Gibco) complete medium;

[0117] 4.2 The predicted peptide sequence is synthesized by solid phase, and the purity of the peptide is ≥95%. It is dissolved in DMSO and stored at -80°C;

[0118] 4.3 Add the following raw materials into the 24-well plate: T2 cells, 1X10^6 cells / well; natural human β2 microglobulin (Prospec), the final concentration is 0.5 μM; the final concentration gradient of each polypeptide is set to: 2.5 μM, 5 μM, 10 μM, 20 μM, 40 μM and 80 μM were added to 24-well plates respectively, and co-incubated for 16 hours in a 5% CO2 incubator at 37°C. The experiment set up a blank group and a control group (without adding peptide);

[0119] 4.4 Transfer the cells to a 1.5ml centrifuge tube, wash twice with 1ml 1XPBS, discard the supernatant;

[0120] 4.5 Add FITC Mouse Anti-Human HLA-A2 (BD Biosciences, Ox...

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Abstract

The invention provides a method for screening a tumor neoantigen based on HLA typing and structure, including: A, obtaining an encoded polypeptide sequence corresponding to a mutant gene of a tumor tissue cell and using the same as a polypeptide collection of a potential antigen; B, obtaining an HLA typing collection whose HLA typing frequency in the yellow population exceeds a specified threshold; performing affinity prediction on the polypeptide collection and the HLA typing collection, and selecting out a polypeptide sequence with an affinity exceeding the specified threshold; C, 3D structurally modeling the HLA in the HLA typing collection and 3D structurally modeling the polypeptide sequence; D, performing molecular docking by using the HLA as a receptor and using the polypeptide sequence as a ligand; E, using the polypeptide sequence corresponding to a score exceeding the specified threshold as a candidate polypeptide sequence of a tumor neoantigen. The screening method of the present application facilitates subsequent targeted experiments based on this, can greatly reduce the number of experiments, and saves time, labor, and cost.

Description

technical field [0001] The invention relates to the field of antigen screening, in particular to a screening method for tumor neoantigens based on HLA typing and structure. Background technique [0002] Tumor vaccine is one of the research hotspots in recent years. Its principle is to introduce tumor antigens into patients in various forms, such as tumor cells, tumor-associated proteins or polypeptides, and genes expressing tumor antigens, so as to overcome the immune system caused by tumors. Suppressive state, enhance immunogenicity, activate the patient's own immune system, induce the body's cellular and humoral immune responses, so as to achieve the purpose of controlling or eliminating tumors. In April 2010, the US Food and Drug Administration (FDA) approved Provenge / sipuleucel-T for the treatment of advanced prostate cancer, making it the first autologous active immunotherapy drug and the first real therapeutic cancer vaccine. Paving the way for the development of othe...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G16B15/30
Inventor 张崇骞赵永浩马赛闫成海张晓霞J·彭D·张
Owner BETA PHARM SUZHOU LTD
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