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Fusion protein of human thymosin alpha1 and human composite interferon and preparation thereof

A technology of fusion protein and function, which is applied in the direction of peptide/protein components, medical preparations containing active ingredients, hybrid peptides, etc., and can solve problems such as reduced affinity, decreased antiviral specific activity, and difficulties in clinical application

Inactive Publication Date: 2004-08-25
重庆康尔威药业股份有限公司
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  • Abstract
  • Description
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  • Application Information

AI Technical Summary

Problems solved by technology

However, it has been found that the fusion of IFN-α with TM-alpha1 at the N-terminal or C-terminal will not cause significant changes in the conformation of the two, but for interferon, it may be due to the existence of steric hindrance after fusion. And reduce its affinity with related receptors, cause antiviral specific activity to drop greatly, when its antiviral specific activity reduces to a certain extent, it will cause difficulties in clinical application.

Method used

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  • Fusion protein of human thymosin alpha1 and human composite interferon and preparation thereof
  • Fusion protein of human thymosin alpha1 and human composite interferon and preparation thereof
  • Fusion protein of human thymosin alpha1 and human composite interferon and preparation thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0068] Synthesis of fusion genes

[0069] Design and Synthesis of Synthetic Fragments

[0070] According to the customary code of Escherichia coli, the IFN-con-Linker-TM-alpha1 fusion protein gene sequence (see figure) was artificially designed, and the whole gene sequence was entrusted to Shanghai Sangon Bioengineering Co., Ltd. to synthesize.

[0071] Construction of Engineering Bacteria for Fusion Gene Expression

[0072] Construction of Fusion Gene Expression Plasmid

[0073] Take the TM-alpha-Linker-IFN-con1 fusion gene plasmid and recover the small fragment after double digestion with endonucleases Nde I and BamH I. The prokaryotic expression plasmid pET-22b(+) with the T7 promoter was passed through Nde I and BamH I. Large fragments were recovered after BamHI double digestion. Ligated by T4 ligase, transformed into Escherichia coli DH5α, and screened recombinants. The clone confirmed by enzyme digestion and sequence analysis was named pET-TIN.

[0074] Construction...

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Abstract

The present invention relates to a fusion protein of human thymosin alpha 1and human compound interferon, and said protein expression and purification method and application of said protein which can be used as medicine for resisting virus and resisting tumor.

Description

Technical field: [0001] The invention relates to a fusion protein of human compound interferon (IFN-con) and thymosin alpha 1 (Thymosin alpha 1) and a method for expressing and purifying the protein and using the protein as an antiviral and antitumor drug. Background technique: [0002] Interferon is a kind of protein polypeptide with relatively small molecular weight composed of single-chain molecules, which has anti-virus, anti-proliferation and immune regulation functions, and is clinically used to treat viral diseases, tumors and multiple sclerosis. Human interferon is divided into three categories, α, β and γ, which are mainly produced by leukocytes, fibroblasts and lymphocytes, respectively. Human alpha interferon consists of 166 or 165 amino acids, with a molecular weight of about 19,000 Daltons. At present, more than 20 human alpha interferon submembers have been cloned and determined for their amino acid sequences. Although interferon preparations have great clinic...

Claims

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Application Information

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IPC IPC(8): A61K38/21A61P31/12A61P35/00C07K19/00C12N15/62C12N15/63
Inventor 黎波刘先俊周辉云邓显峰
Owner 重庆康尔威药业股份有限公司
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