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A kind of human muc1 specific binding polypeptide and its extraction method

A technology that combines peptides and extraction methods, applied in the field of peptide extraction and separation, and protein, can solve the problem of weak drug specificity, and achieve the effects of easy chemical synthesis, good application value, and low immunogenicity.

Active Publication Date: 2022-07-05
HUBEI UNIV OF ARTS & SCI
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  • Abstract
  • Description
  • Claims
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Problems solved by technology

[0009] The present invention mainly aims at the problem that the specificity of existing tumor diagnosis and treatment drugs is not strong. Taking human MUC1 protein, a classic tumor marker, as the target molecule, a polypeptide sequence is obtained by biological panning of 12 random peptide libraries presented by phage

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  • A kind of human muc1 specific binding polypeptide and its extraction method

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Embodiment 1

[0042] The human MUC1 specific binding polypeptide provided in this application is extracted and obtained by the following method:

[0043] S1. Bio-panning: add MUC1 solution to the well plate, inoculate E. coli strain and medium containing tetracycline, and cultivate; pour off the residual liquid, add TBS blocking solution to wash; add phage solution to amplify, purify, and elute to obtain The eluted phage; the eluted phage solution is added to the E. coli culture for amplification, and the amplified eluate is separated; the second and third rounds of screening are repeated to obtain the screened positive plaques;

[0044] The specific steps of the above step S1 are:

[0045] S11. Add 200ul NaHCO with a concentration of 100ug / mL and 0.1mol / L NaHCO to each well of the 96-well plate 3 The diluted human MUC1 protein solution was placed in a wet box at 4°C with gentle shaking and incubated overnight; 10mL of Escherichia coli ER2738 was inoculated into 20mL of LB medium containin...

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Abstract

The invention discloses a human MUCl specific binding polypeptide, the amino acid sequence of which is HKFIPHRDTSIA. The polypeptide is verified by methods such as biopanning, amplification of positive phage, DNA sequence determination of the polypeptide encoded by the phage, and ELISA assay on the screened positive phage. The polypeptide has small molecular weight, easy chemical synthesis, low immunogenicity, can specifically bind to human MUC1 protein, and has good application value in the field of tumor diagnosis and treatment targeting human MUC1 protein.

Description

technical field [0001] The invention belongs to the technical field of protein and polypeptide extraction and separation, in particular to the extraction technology of human MUC1 specific binding polypeptide. Background technique [0002] Tumor is one of the diseases that seriously threaten human health. With the deepening of people's understanding of tumor and the continuous development of science and technology, clinical drugs for tumor diagnosis and treatment continue to appear, but how to effectively improve tumor diagnosis and treatment drugs so far? The specificity of tumor diagnosis and the reduction of the combination of tumor diagnosis and treatment drugs with non-tumor tissues is still a problem that restricts tumor diagnosis from entering the stage of precision. Targeted drug strategy is an effective method to solve this problem. This method uses molecules (such as receptors and other molecules) that are highly expressed in tumor tissues compared with normal tissu...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K7/08C12N15/10C40B30/04
CPCC07K7/08C12N15/1037C40B30/04Y02A50/30
Inventor 秦鑫陶炳灼贺子涵许世琦张文静刘倩蓉
Owner HUBEI UNIV OF ARTS & SCI
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