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Four-branch polypeptide and application thereof

A four-branch, cell-based technology, applied in the field of life sciences, can solve the problems that limit the widespread application of tumor vaccines, and achieve the effects of improving the body's immunogenicity, broad clinical application prospects, and prolonging the half-life

Inactive Publication Date: 2016-04-20
深圳市中美康士生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, most of the tumor-associated antigens discovered so far are tissue-specific, such as AFP, which only targets liver cancer, etc. Immunotherapy with these antigens can only work on the corresponding tumors, which limits the wide application of tumor vaccines targeting these antigens

Method used

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  • Four-branch polypeptide and application thereof
  • Four-branch polypeptide and application thereof
  • Four-branch polypeptide and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Synthesis and determination of purity and molecular weight of quarter polypeptide

[0029] 1. The synthesis of peptides was carried out on a peptide synthesizer (ABI43IA). According to the above target polypeptide sequence, the peptide chain is extended one by one from the carboxyl terminal to the amino terminal.

[0030] 2. Add the synthesized peptide resin into the cutting solution (ice bath), stir the reaction to crack the peptide chain from the branch, and remove various protective groups at the same time.

[0031] 3. After filtering and evaporating, vacuumize the resulting crude peptide and store it in a -20°C refrigerator.

[0032] 4. Dissolve the polypeptide, adjust the concentration to 10 mg / ml, and filter with a 0.45 μm fiber membrane. Purification using medium pressure liquid chromatography. Different filler gel columns are required for purification: SOURCE-30-RPC filler purification column is suitable for water-soluble peptides, while fat-soluble peptides ...

Embodiment 2

[0038] The killing experiment of CTL induced by the tetrabranched peptide of the present invention and the measurement of IFN-γ secretion

[0039] In the initial culture medium of PBMC, add DC cell activation factor GM-CSF (140ng / ml), IL-4 (50ng / ml), TNF-α (50ng / ml) and polyI:C (25ug / ml) and The four-branched polypeptide prepared in Example 1 and the corresponding single peptide (0.1ug / ml / each) were added, placed in a 37°C, 5% CO2 incubator for 12 days, and harvested.

[0040] In order to verify the universality of the four-branched polypeptide, the four-branched polypeptide and the corresponding single peptide were respectively loaded on DC, and the killing effect of the induced CTL on tumor cells from different tissues was studied. as attached image 3 As shown, the tetrabranched peptide of the present invention and its corresponding monopeptide can induce specific CTL responses, and it has been detected that it has obvious killing effects on K562 and A549. At the maximum ...

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Abstract

The invention provides a synthetic functional four-branch polypeptide (AA1-AA2)2-(Lys-NH2)2-Lys-SER-NH2, wherein AA1 and AA2 are 15-AA peptides of SOX2 and OCT4 respectively. The branched synthetic polypeptide product provided by the invention has the effects of comprehensively improving the immunogenicity of an organism and prolonging the half life of the organism, and has broad application prospect in preparation of an anti-tumor medicament or vaccine.

Description

technical field [0001] The invention relates to the technical field of life sciences, in particular to a four-branched polypeptide and its application. Background technique [0002] The incidence of tumors has been on the rise since cancer was recorded, especially in the past two or three decades, the incidence of tumors has been increasing at a rate of 3% to 5% per year, and 80% of new tumors occur in the developing countries. At present, there are about 13 million new tumors every year in the world, and 8 million deaths in that year. In my country, there are about 2.5 million new patients and nearly 1.8 million deaths each year. Recent epidemiological surveys show that cancer has become the number one cause of death in major large and medium-sized cities in my country. Surgery, radiotherapy, and chemotherapy are still the main means of cancer treatment, but surgery has poor curative effect on patients with advanced disease, and radiotherapy and chemotherapy show the limi...

Claims

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Application Information

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IPC IPC(8): C07K19/00C07K14/435C12N15/62C12N5/0783A61K38/17A61K39/00A61K48/00A61P37/04A61P35/00
CPCA61K38/00A61K39/00A61K39/0011C07K14/435C07K2319/00C12N5/0638
Inventor 佘超然刘根桃李晓祥
Owner 深圳市中美康士生物科技有限公司
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