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Method for screening polypeptide specifically bound with human colon cancer cells and application of polypeptide

A colon cancer cell and peptide-binding technology, applied in the medical field, can solve the problems of high price, low penetrating power, strong immunogenicity, etc., achieve the effect of small molecular weight and overcome immune allergic reaction

Active Publication Date: 2017-07-21
孟祥军
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The curative effect of metastatic colon cancer surgery is poor, the recurrence rate is high after surgery, and the 5-year survival rate of patients with unresectable tumors is almost zero
Although drug-targeted therapy has achieved great success, there are still many deficiencies such as the large molecular weight of the antibody used for targeting, low penetration in tumor tissue, and toxic effects on the liver and bone marrow; strong immunogenicity; preparation The process and cycle are long, expensive, etc.

Method used

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  • Method for screening polypeptide specifically bound with human colon cancer cells and application of polypeptide
  • Method for screening polypeptide specifically bound with human colon cancer cells and application of polypeptide
  • Method for screening polypeptide specifically bound with human colon cancer cells and application of polypeptide

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Embodiment 1

[0020] 1. Screening method:

[0021] The screening method for specific binding polypeptides on the surface of colon cancer cells in this embodiment includes the following steps:

[0022] Step (1): Use the whole cell screening method to incubate the random dodecapeptide library phage clones with colon cancer cells, discard the clones that have not combined with the colon cancer cells, and combine them with the colon cancer cells. Amplification of combined clones;

[0023] Step (2): Repeat step (1) 4 times to maximize the enrichment of phage clones that bind to the colon cancer cells to obtain clones that have a higher affinity for colon cancer cells, and then expand;

[0024] Step (3): Incubate the amplified clones with normal colonic epithelial cells, perform negative screening, and remove non-specific clones that bind to normal cells.

[0025] Step (4): sequence the cloned strain after the final amplification, and analyze the polypeptide sequence.

[0026] The polypeptide sequence has ...

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Abstract

The invention relates to a polypeptide specifically bound with the surfaces of colon cancer cells and application of the polypeptide. A method for screening the polypeptide includes steps of (1), incubating clone strains with amplified bacteriophage dodecapeptide libraries and the colon cancer cells together, discarding the clone strains which are not bound with the colon cancer cells, screening the clone strains and then amplifying the screened clone strains which are bound with the colon cancer cells; (2), repeatedly carrying out the step (1) by four times, continuously enriching bacteriophage clone strains to obtain clone strains with high colon cancer cell affinity and repeatedly verifying the binding capacity of the bacteriophage clone strains in liquid phases; (3), incubating the bacteriophage clone strain obtained at the step (2) and normal colonic epithelial cells and removing non-specifically bound clone strains; (4), carrying out sequencing on positive clone strains; (5), verifying the specificity of the polypeptide on the clone cancer cells and tissues by the aid of immunofluorescence. The polypeptide and the application have the advantages that short peptides which are specifically bound with the colon cancer cells are screened by the aid of technologies for displaying peptide libraries by the aid of bacteriophage, and the polypeptide can be used as a molecular marker or targeted carrier or the like for targeted therapeutic medicines.

Description

Technical field [0001] The present invention relates to the field of medicine, in particular to a method for screening colon cancer cells with specific binding polypeptides and applications thereof. Background technique [0002] Colon cancer is one of the common malignant tumors of the digestive system. Its incidence rate is second only to gastric cancer and esophageal cancer. It seriously affects human life and health. Currently, 440,000 new cases of colorectal cancer are diagnosed in my country each year, and as many as 230,000 people die each year. . Early detection and early removal of precancerous lesions and focal cancer can effectively reduce the mortality of colorectal cancer. However, due to the slow development of colon cancer, the early symptoms are often not obvious. 6% of asymptomatic people over 40 have early colon cancer, 70%-80% of colon cancer patients have entered the middle and advanced stages when they are diagnosed, and more than 25% of patients have cancer c...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K7/08A61K38/10A61P35/00G01N33/68
CPCA61K38/00C07K7/08G01N33/6818
Inventor 孟祥军田晓慧侯利丹祝丹希梁钰王建华
Owner 孟祥军
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