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Screening method and application of human colon cancer cell specific binding polypeptide

A technology of combining human colon cancer cells and peptides, which is applied in the medical field, can solve problems such as low penetration, high price, and toxic effects, and achieve the effect of overcoming immune allergic reactions and small molecular weight

Active Publication Date: 2020-06-16
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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  • Screening method and application of human colon cancer cell specific binding polypeptide
  • Screening method and application of human colon cancer cell specific binding polypeptide
  • Screening method and application of human colon cancer cell specific binding polypeptide

Examples

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Embodiment 1

[0019] 1. Screening method:

[0020] The screening method for specific binding polypeptides on the surface of colon cancer cells in this embodiment includes the following steps:

[0021] Step (1): Using the whole-cell screening method, incubate the random twelve-peptide library phage clones with colon cancer cells, discard the clones that are not combined with the colon cancer cells, and combine with the colon cancer cells Expansion of bound clones;

[0022] Step (2): Cycle step (1) 4 times to maximize the enrichment of phage clones that bind to the colon cancer cells to obtain clones that bind to colon cancer cells with a higher affinity and perform amplification;

[0023] Step (3): Incubating the above-mentioned amplified clones with normal colonic epithelial cells, and performing negative selection to remove non-specific clones combined with normal cells.

[0024] Step (4): Perform sequence determination on the final amplified clone, and analyze the polypeptide sequence. ...

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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 invention relates to the field of medicine, in particular to a screening method for specific binding polypeptides on the surface of colon cancer cells and its application. 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. At present, there are 440,000 newly diagnosed cases of colorectal cancer in my country every year, and as many as 230,000 patients die every year. . Early detection and early resection of precancerous lesions and focal cancer can effectively reduce the mortality rate of colorectal cancer. However, due to the slow development of colon cancer, the early symptoms are not obvious. 6% of asymptomatic people over the age of 40 suffer from early colon cancer, 70%-80% of colon cancer patients have entered the middle and late stages when they are diagnosed, and more t...

Claims

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Application Information

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Patent Type & Authority Patents(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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