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Application of extracellular segment of low pH-value insertion peptide as antigen

A technology of extracellular segments and antigens, applied in the field of biomedicine, can solve the problems of unreported low pH insertion peptide antigen properties

Active Publication Date: 2020-01-21
BEIJING ZEQIN BIOMEDICAL CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] However, the prior art has not reported that the low pH insertion peptide has antigenic properties, and the application of using antigenic properties to prepare antibodies

Method used

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  • Application of extracellular segment of low pH-value insertion peptide as antigen
  • Application of extracellular segment of low pH-value insertion peptide as antigen

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0064] Example 1 Preparation of Antibodies Using var-7 and var-3 Low pHLIP Insertion Peptide Extracellular Parts as Antigens

[0065] 1. Experimental materials

[0066] MC38 cells were purchased from ATCC;

[0067] Var-7 low pHLIP insertion peptide and var-3 low pHLIP insertion peptide were synthesized by Beijing Huada Protein Research and Development Center Co., Ltd., dissolved in PBS at a concentration of 40 μM;

[0068] 6-8 week-old female C57 / BL6 mice were purchased from Victoria Lihua.

[0069] 2. Antibody preparation

[0070] (1) The extracellular segment connects to KLH

[0071] The extracellular segment of Var-7 low pHLIP insertion peptide (Ala-Cys-Glu-Glu-Gln-Asn-Pro, SEQ ID NO.18) is connected to KLH (synthesized by Hangzhou Zhongpei Biochemical Co., Ltd.), var-3 low pHLIP The extracellular segment of the inserted peptide (Ala-Cys-Asp-Asp-Gln-Asn-Pro, SEQ ID NO.19) was connected to KLH (synthesized by Hangzhou Zhongpei Biochemical Co., Ltd.).

[0072] (2) Animal...

Embodiment 2

[0074] Example 2 Anti-tumor Effect Evaluation of Antibodies

[0075] 1. Experimental steps

[0076] MC38 was inoculated into C57 / BL6 mice. When the tumor diameter reached 1cm, the tumor was aseptically peeled off, chopped, homogenized, filtered, and made into a single cell suspension, cultured and expanded in 1640 complete medium, and the cells were injected Subcutaneously on the flank of C57 / BL6 mice, 2x10 6 cells / mouse, when the diameter of the tumor grows to 0.8-1cm, remove the tumors that are too large and too small, and group the mice with tumors that are basically the same size. Divided into 4 groups: var-7 / 3 single injection group, 10 rats; var-7 / var-3 combined with 3D1 / 4D3 injection group, 10 rats; 3C1 / 4D3 single injection group, 10 rats; normal saline N.S injection group, 10 only. Tumor size was measured every 3 days.

[0077] Administration method: var-7 / 3 administration: intravenous injection of 40 μM / 100 μl per animal, starting from the day after the grouping i...

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Abstract

The invention discloses application of an extracellular segment of a low pH-value insertion peptide as an antigen. Research of the invention finds that the extracellular segment of the low pH-value insertion peptide has antigenicity, an antibody can be prepared by using the extracellular segment of the low pH-value insertion peptide as the antigen, and the antibody can be used in tumor treatment.The research results of the invention provide novel candidate drugs for clinical tumor treatment.

Description

technical field [0001] The invention belongs to the field of biomedicine and relates to a low-pH insertion peptide, and also relates to the application of the extracellular part of the low-pH insertion peptide as an antigen. Background technique [0002] The low pH insertion peptide (pHLIP, pH low insertion peptide) derived from bacteriorhodopsin transmembrane helix protein C is a water-soluble polypeptide that can insert into the cell bilayer lipid membrane to form a stable transmembrane α-helix. Peptide folding and membrane insertion are driven by a pH drop from neutral or basic (pH > 7.4) to mildly acidic (pH = 7.0-6.5 or lower). pHLIP has three main forms: form I, which is unstructured and soluble in water at neutral pH, state II, which is unstructured and bound to the cell membrane surface at neutral pH, and intercalates at acidic pH and crosses the cell membrane as an alpha helix State III. Because the poor solubility caused by easy aggregation is a property of me...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/195C07K19/00C07K16/12C12N15/31G01N33/569A61K39/40A61P35/00
CPCA61K2039/505A61P35/00C07K14/195C07K16/12C07K2319/00G01N33/56911
Inventor 魏化伟杨承刚
Owner BEIJING ZEQIN BIOMEDICAL CO LTD