Tumor neoantigen epitope peptide and its application

A tumor antigen and epitope peptide technology, applied in the field of molecular immunology, can solve the problems of unsatisfactory survival and achieve the effect of preventing recurrence and metastasis

Active Publication Date: 2021-03-26
中生康元生物科技(北京)有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Surgical treatment adjuvant local or systemic chemotherapy, the survival of most cancer patients is not satisfactory

Method used

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  • Tumor neoantigen epitope peptide and its application
  • Tumor neoantigen epitope peptide and its application
  • Tumor neoantigen epitope peptide and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0145] Example 1 Prediction of Tumor Neoantigen Epitope Peptides

[0146] 1. Material preparation

[0147] The tumor tissues of tumor patients were obtained, and WES and RNA-seq sequencing of tumor tissues were completed through the illumina high-throughput sequencing platform.

[0148] 2. Data quality control

[0149] The original fastq data of DNA and RNA sequencing were quality controlled by FastQC software to obtain the data clean.fq.gz after quality control filtering.

[0150] 3. Data comparison

[0151] The DNA data after quality control was compared and analyzed using BWA software to obtain bam files of tumor and normal tissue DNA data, and the RNA after quality control was compared and analyzed using hisat2 software to obtain bam files of tumor RNA data.

[0152] 4. Bam file processing

[0153] The bam files after comparison need further processing. The bam files of DNA data are sorted by using samtools, Picard, GATK and other software to sort the bam files, remove...

Embodiment 2

[0171] Example 2 Detection of Neoplastic Antigen Epitope Peptide and Tetramer Binding Ability

[0172] Tumor neoantigens to stimulate T cell activation must first be able to combine with HLA molecules of antigen-presenting cells (APCs) to form a complex expressed on the cell surface for recognition by T cell receptors on the surface of T cells. Therefore, recombinant tetramers were used to test the binding ability of predicted epitope peptides to tetramers.

[0173] The detection method is based on the ability of MHC class I molecules to replace peptides. MHC class I tetramers are formed from monomer units folded by a weak affinity peptide. Under the action of a suitable peptide replacement factor, it can produce Tetrameric complex that binds a specific peptide of interest. The detection technology is based on flow cytometry, and the main reagents include antibody-coupled magnetic beads that can capture MHC class I tetramers, and FITC-labeled antibodies that can recognize the...

Embodiment 3

[0186] Example 3. Analysis of cellular immune responses induced by tumor neoantigen epitope peptides

[0187] DC cells are loaded with neoantigen epitope peptides that can bind to HLA molecules and then injected into tumor patients. The frequency of antigen-specific T cells that produce γ-interferon is detected by ELISPOT experiment, and the tumor patients are analyzed after inoculation of neoantigen-loaded DC vaccines. Antigen-specific T lymphocyte responses.

[0188] 3.1 Elispot detection of specific cytotoxic T cells in peripheral blood of patient A

[0189] Whole blood was collected from patient A, and after the plasma was separated, human peripheral blood mononuclear cells (PBMCs) were isolated according to GE standard (GE Healthcare Life Sciences, Ficoll) operating procedures, and counted and viability were performed. Take a certain amount of PBMC to prepare a cell density of 2×10 6 / ml solution, then add the epitope peptide obtained in Example 2 into PBMC (the volume ...

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Abstract

The invention discloses a tumor neoantigen epitope peptide and its application. The epitope peptide comprises one or more than two amino acid sequences selected from those shown in SEQ ID NO: 1-6. The epitope peptide can be combined with MHC molecules on human cells, and the combined dendritic cells can stimulate the activation and expansion of T lymphocytes with tumor cell killing effect. Therefore, the dendritic cells combined with the epitope peptide can be It is used for the treatment of tumors, and the prevention of recurrence and metastasis of tumor patients after surgery.

Description

technical field [0001] The invention relates to the technical field of molecular immunology, in particular to a tumor neoantigen epitope peptide and its application. Background technique [0002] The occurrence of tumors in the body is a multi-stage and multi-step complex process, accompanied by changes in cytology and genetics, various gene mutations and abnormal gene expression, including the activation and / or inhibition of oncogenes. Oncogene inactivation stimulates abnormal signal transduction pathways, leading to abnormal cell cycle and apoptosis, resulting in malignant transformation of cells, and with the accumulation of various molecular abnormalities, tumor progression and metastasis occur. Therefore, understanding these genes specifically expressed in tumor tissue during the process of tumorigenesis is of great significance for the elucidation of tumorigenesis and development mechanism, clinical diagnosis and treatment. [0003] The incidence and mortality of tumo...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/47C07K19/00C12N15/12C12N15/62C12N5/10A61K39/00A61K39/385A61P35/00A61P35/04
CPCA61K39/0011A61K39/385A61K2039/5154A61K2039/6031A61P35/00A61P35/04C07K14/4748C07K2319/10C07K2319/40C12N5/0639C12N2510/00
Inventor 程旭东尹艳慧李欢欢
Owner 中生康元生物科技(北京)有限公司
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