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A screening method for tumor neoantigens based on hla typing and structure

An antigen and tumor technology, applied in the analysis of two-dimensional or three-dimensional molecular structure, bioinformatics, instruments, etc., can solve the problems of high cost, time-consuming, labor-intensive, etc., to save money, reduce the number of experiments, and save labor and expenses. Effect

Active Publication Date: 2022-04-12
BETA PHARM SUZHOU LTD
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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

Method used

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  • A screening method for tumor neoantigens based on hla typing and structure
  • A screening method for tumor neoantigens based on hla typing and structure
  • A screening method for tumor neoantigens based on hla typing and structure

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Embodiment 1

[0050] like 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 am...

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 present invention provides a method for screening tumor neoantigens based on HLA typing and structure, comprising: A, obtaining the polypeptide sequence encoded by the mutant gene of tumor tissue cells, and using it as a polypeptide set of potential antigens; B, Obtain the HLA typing set whose frequency of HLA typing in the yellow population exceeds a specified threshold; perform affinity prediction between the polypeptide set and the HLA typing set, and select polypeptide sequences whose affinity exceeds the specified threshold; C. HLA typing Three-dimensional structure modeling of HLA in the collection; and three-dimensional structure modeling of polypeptide sequences; D. Using HLA as a receptor and polypeptide sequences as ligands for molecular docking; E. The corresponding polypeptide sequence when the score exceeds the specified threshold Candidate polypeptide sequences as tumor neoantigens. From above, through the screening method of the present application, it is convenient to conduct further targeted experiments based on it, which can greatly reduce the number of experiments, and save time, effort and money.

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 Patents(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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