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A liver cancer microsphere cell surface specific targeting polypeptide, and a preparing method and applications thereof

A cell surface and peptide-targeting technology, which is applied to peptides, instruments, and analytical materials, can solve the problems of complex structure, large molecular weight, and high cost of tumor cells

Inactive Publication Date: 2017-07-14
GUIZHOU MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Antibodies, as targeting ligands for the diagnosis and treatment of cancer, are difficult to penetrate tumor cells wrapped in a dense matrix and have high cost, large molecular weight, complex structure, and immunogenicity.

Method used

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  • A liver cancer microsphere cell surface specific targeting polypeptide, and a preparing method and applications thereof
  • A liver cancer microsphere cell surface specific targeting polypeptide, and a preparing method and applications thereof
  • A liver cancer microsphere cell surface specific targeting polypeptide, and a preparing method and applications thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0185] Example 1 A specific targeting polypeptide on the cell surface of liver cancer microspheres, the polypeptide is a heptapeptide, and its amino acid sequence is Asn-Thr-Gly-Ser-Pro-Tyr-Glu.

Embodiment 2

[0186] Example 2 The specific targeting polypeptide on the cell surface of liver cancer microspheres is a cyclic heptapeptide with a structural formula: cyclo-(Asn-Thr-Gly-Ser-Pro-Tyr-Glu).

Embodiment 3

[0187] Example 3 A method for preparing a specific targeting polypeptide on the cell surface of liver cancer microspheres, comprising the following steps:

[0188] (1) Culture of adherent cells and liver cancer microspheres: Liver cancer microspheres, HCCLM3 cells and LO2 cells were cultured according to conventional methods;

[0189] (2) Liver cancer microspheres cultured in HCCLM3 cells in suspension were used as target cells, and HCCLM3 cells and LO2 cells cultured on the adherent wall were used as adsorbed cells. Phage display technology was used for differential screening. The bottom LO2 cells were washed 3 times with PBS, added with BSA, blocked for 30 min in a constant temperature culture shaker at 37°C and 50 rpm, and then washed 3 times with PBST buffer; the original phage peptide library solution was drawn and mixed with PBS Mix well to obtain a mixed solution; add the mixed solution to adherent LO2 cells, and incubate in a constant temperature culture shaker at 37°C...

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Abstract

The invention discloses a liver cancer microsphere cell surface specific targeting polypeptide. The polypeptide is a heptapeptide, and the amino acid sequence of the polypeptide is Asn-Thr-Gly-Ser-Pro-Tyr-Glu. A preparing method and applications of the polypeptide are also disclosed. The polypeptide can be specifically bound with liver cancer microsphere cells, has high targeting binding capacity for liver cancer microspheres, high affinity and high specificity, and has no influence on cell growth. According to the method, liver cancer microspheres subjected to HCCLM3 cell suspension culture are adopted as target cells, HCCLM3 cells and LO2 cells subjected to adherent culture are adopted as adsorption cells, and subtractive screening is performed by utilizing a phage display technology. In the method, storage capacity is high, the screening success rate is high, operation is simple and the cost is low. The screened polypeptide can be used for preparing liver cancer embolus diagnostic reagents or liver cancer targeting treatment medicines.

Description

technical field [0001] The invention relates to a specific targeting polypeptide on the cell surface of liver cancer microspheres and its preparation method and application, specifically belonging to the field of biotechnology. Background technique [0002] The clinical incidence of HCC with cancer embolus is very high, and the prognosis is often poor, making diagnosis and treatment very difficult. Studies suggest that liver cancer tumor thrombi are induced by hepatic cancer stem cells (HCSC) in the portal vein or bile duct microenvironment, while retaining some of the original differentiation characteristics of stem cells. Hepatoma Microspheres is an ideal model for the study of liver cancer stem cells, and also exhibits the characteristics of tumor thrombi, making it an ideal model for the study of tumor thrombi. Targeting tumor thrombi for diagnosis and treatment is the current direction of clinical efforts for liver cancer with tumor thrombi, and finding molecules that ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K7/06C07K7/64G01N33/574
CPCC07K7/06C07K7/64G01N33/57438
Inventor 刘扬陈腾祥洪阳张金娟王晋星一陈妮刘伟兰金芝朱晨蕾孙密欣唐佳艳王寒琪郭蔓岚曾志锐雷珊严莉莎钟钰徐澍毛大华杨琼
Owner GUIZHOU MEDICAL UNIV
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