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Polypeptide specifically combined with HepG2 cell surface

A cell surface and specific technology, applied in the direction of peptides, etc., can solve the problems of low early detection rate of liver cancer and high patient mortality

Inactive Publication Date: 2012-07-04
SHAANXI NORMAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the lack of specific, cheap and convenient early diagnosis methods for liver cancer at present, the early detection rate of liver cancer is still very low, and the mortality rate of patients is still high.

Method used

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  • Polypeptide specifically combined with HepG2 cell surface
  • Polypeptide specifically combined with HepG2 cell surface
  • Polypeptide specifically combined with HepG2 cell surface

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Experimental program
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Embodiment Construction

[0020] 1. Technical route

[0021] The present invention adopts the phage polypeptide display technology, takes the human liver cancer HepG2 cell line as the target cell, and uses the human embryonic kidney HEK293 cell as the negative adsorption cell to carry out four rounds of subtractive screening, and screens the polypeptide that can specifically bind to the liver cancer HepG2 cell from the phage 12 peptide library Motif, retrieve the cell surface receptors it may recognize, and provide a good experimental basis for further research on early diagnosis and targeted therapy of liver cancer. The specific technical route is as follows: figure 1 shown.

[0022] 2. Materials and methods

[0023] 2.1 Main experimental materials

[0024] 2.1.1 Cells, phage peptide library, host bacteria

[0025] (1) Cell lines: human liver cancer cell line HepG2, human embryonic kidney cell line HEK293, purchased from ATCC (Rockville, USA).

[0026] (2) Phage peptide library: random dodecapepti...

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Abstract

The invention discloses a polypeptide specifically combined with a HepG2 cell surface. According to the invention, four polypeptide segments are selected by utilizing a phage display random dodecapeptide library, the amino acid sequences of the four polypeptide segments are respectively LLADTTHHRPWT, LLADTPHHRPWT, FGWVTPHHELRS and SLSDLTHMGPWP. According to the invention, a polypeptide sequence combined with a liver cancer HepG2 cell is selected by utilizing a phage polypeptide display technology, and ELISA (enzyme-linked immunosorbent assay) identifies the affinity of phage clone and the liver caner cell, thus eight phage clones are obtained; four polypeptide sequences are obtained by sequencing, wherein the common amino acid sequence (basic sequence) is ***D(V)TT(P)HH*P(L)W(R)*; homology analysis indicates that the basic sequence of the polypeptide is possibly amino acid determinant on a ligand protein combined with a tumor cell surface receptor; cell immunofluorescence further identities that the target result of the positive clone of the phage prompts that the positive clone of the phage can be specifically combined with the HepG2 cell; and the selected specific polypeptide of the liver cancer HepG2 cell provides an experiment basis for early diagnosis of liver cancer, targeting delivery of an antitumor medicine and research and development of a targeting short peptide medicine.

Description

technical field [0001] The invention relates to the field of biotechnology, and relates to screening tumor tissue-specific binding peptides, in particular to a polypeptide specifically binding to the surface of HepG2 cells. Background technique [0002] Liver cancer (Hepatocarcinoma) is one of the common malignant tumors of the digestive tract. In China, liver cancer is a disease with high morbidity, high mortality and high treatment costs. According to the data of the World Health Organization, there are at least 550,000 deaths from liver cancer in my country every year. More than 80% of liver cancer patients are transformed from hepatitis virus carriers or chronic hepatitis patients. my country is one of the high-incidence areas of liver cancer in the world, and its incidence rate of liver cancer is about 10 times that of Europe and the United States, mainly distributed in the southeast coast. The number of new cases of such tumors in developing countries accounts for a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K7/08
Inventor 侯颖春尹田乐
Owner SHAANXI NORMAL UNIV
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