Antigen peptide composition and application thereof

An antigen epitope and peptide composition technology, applied in the fields of genetic engineering and biomedicine, can solve problems such as insufficient curative effect, and achieve the effects of simple and easy processing, convenient synthesis, and enhanced ability

Active Publication Date: 2017-04-26
广东龄值生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] At present, the preparation of DC vaccines is mostly based on single-antigen epitope peptide loading targeting conventional tumor markers, but single-antigen peptide-loaded DC cells targeting tumor cells are not effective in tumor treatment

Method used

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  • Antigen peptide composition and application thereof
  • Antigen peptide composition and application thereof
  • Antigen peptide composition and application thereof

Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0034] The preparation method step of described artificial synthesis is:

[0035] First, the antigenic peptide STEAP1 (SEQ ID NO: 1) is selected from polypeptides restricted by allele HLA-A2 sites 292-300 of the STEAP1 molecule. The antigenic peptide PSMA (SEQ ID NO: 2) is selected from polypeptides restricted by allele HLA-A24 positions 178-186 of the PSMA molecule. The antigenic peptide DKK1 (SEQ ID NO: 3) is selected from polypeptides restricted by allele HLA-A2 sites 20-29 of the DKK1 molecule. The antigenic peptide CD133 (SEQ ID NO: 4) is selected from polypeptides restricted by allele HLA-A2 sites 753-761 of the CD133 molecule.

[0036] The present invention entrusts Nanjing GenScript Biotechnology Co., Ltd. to synthesize the antigenic peptide STEAP1 (SEQ ID NO: 1), the antigenic peptide PSM (SEQ ID NO: 2), the antigenic peptide DKK1 (SEQ ID NO: 3) and the The above antigenic peptide CD133 (SEQ ID NO: 4), and its sequence and molecular weight were identified by high pe...

Embodiment 1

[0062] Example 1 DC cell acquisition

[0063] 1) Collect 50ml of peripheral blood, add ficoll (polysucrose) and perform density gradient centrifugation to obtain mononuclear cells.

[0064] 2) Collect the cells in a 6-well plate, count and dilute to 3-5×10 with AIM-V medium 6 / ml, placed in a carbon dioxide incubator for cultivation.

[0065] 3) At 37°C, 5% CO 2 After incubating in the incubator for 90 min, the 6-well plate was gently taken out, and the supernatant and suspended cells in the 6-well plate were sucked out into another culture dish for inducing and culturing T lymphocytes.

[0066] 4) AIM-V medium, rhGM-CSF 800IU / ml, IL-4 500IU / ml were added to adherent cells, and cytokines were added once after 48 hours.

[0067] 5) Harvest the immature DCS cells on the fifth day, add TNF-α250IU / ml on the fifth day of culture, and continue to culture until the eighth day to obtain mature DCs. The antigenic epitope peptide composition is cultured with the mature DCs. During the...

Embodiment 2

[0068] Example 2 Epitope Peptide Loaded DC and Phenotype Identification

[0069] 1) Collect 3×10 immature DCs cultured to the fifth day 6 One, using the antigenic epitope peptide composition provided by the present invention and the single epitope peptide in the composition (antigenic peptide STEAP1, antigenic peptide PSM, antigenic peptide DKK1 and antigenic peptide CD133) at 37 ℃, 5% CO 2 Load in the incubator for 2h.

[0070] 2) Continue culturing in the complete medium until the eighth day, and collect mature DCs loaded with the antigen peptide composition.

[0071] 3) Resuspend in normal saline to a concentration of 5×10 6 individual / ml, human serum albumin was added to a final concentration of 2%, and a DC vaccine loaded with an antigenic epitope peptide composition was prepared.

[0072] 4) DC phenotype changes after loading were detected by flow cytometry.

[0073] 5) DC cells loaded with antigen epitope peptide composition are stained by CD86-PE, CD80-PE, CD40-FIT...

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Abstract

The invention relates to the technical field of genetic engineering and biomedicine, in particular to an antigen peptide composition and application thereof. The provided antigen peptide composition comprises four antigen peptides, namely the antigen peptide STEAP1, the antigen peptide PSM, the antigen peptide DKK1 and the antigen peptide CD133. The experimental results show that when the antigen peptide composition is adopted, dendritic cell maturation can be effectively induced, the antigen presentation ability is improved, specific cytotoxicity killer cells can be generated, the tumor killing efficiency is improved, the growth of tumors can be effectively inhibited, an anti-tumor effect is achieved, and a new selection is provided for the clinic treatment of the prostatic cancer.

Description

technical field [0001] The invention relates to the technical fields of genetic engineering and biomedicine, in particular to an antigenic epitope peptide composition and its application. Background technique [0002] Prostate cancer is the sixth most common cancer in the world and the second most common cancer in men worldwide. The incidence of prostate cancer in European and American countries is higher than that in Asian countries, and it is one of the most common malignant tumors in men, with an incidence rate of more than 100 / 100,000. Although the incidence of prostate cancer in my country is much lower than that in European and American countries, with the increase of life expectancy and the westernization of lifestyle, the incidence and mortality of prostate cancer in my country show a trend of increasing year by year. According to the statistics of cancer in China in 2015, the incidence of prostate cancer in men in my country has reached 60.3 / 100,000, and it may bec...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/00A61P35/00C12N5/0784
CPCA61K39/0005A61K2039/5154A61K2039/5158C12N5/0639
Inventor 马颖李陶马铎阮雯莉陈祥黄婉娴
Owner 广东龄值生物科技有限公司
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