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Fusion protein of tumour new-born blood vessel-specific bonding polypeptide and recombinant human Tum-5, and its preparation method

A technology of tumor neovascularization and binding polypeptide, which is applied in the field of tumor neovascularization specific binding polypeptide and recombinant human Tum-5 fusion protein and its preparation, which can solve the problem of no anti-tumor effect

Inactive Publication Date: 2008-05-28
FOURTH MILITARY MEDICAL UNIVERSITY
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The antigenic epitope of Tumstatin is located at the N-terminal 1-40 amino acids, and this sequence has no anti-tumor effect

Method used

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  • Fusion protein of tumour new-born blood vessel-specific bonding polypeptide and recombinant human Tum-5, and its preparation method
  • Fusion protein of tumour new-born blood vessel-specific bonding polypeptide and recombinant human Tum-5, and its preparation method
  • Fusion protein of tumour new-born blood vessel-specific bonding polypeptide and recombinant human Tum-5, and its preparation method

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Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0063] 1. Construction of prokaryotic expression vector of Tum-5 and Tum-5-NGR His fusion gene

[0064] 1.1 Construction of prokaryotic expression vector of Tum-5His fusion gene

[0065] According to the human Tum-5 gene sequence published in GenBank, the codons encoding Tum-5 amino acids were replaced with codons commonly used in E. coli (which is conducive to the efficient expression of the target protein in E. coli) and handed over to Shanghai Boya Biotechnology Co., Ltd. The company synthesizes the entire gene, introduces the SphI restriction site upstream, and introduces the HindIII restriction site downstream, and clones the target gene into the vector pMD-18T, named pMD-18T-Tum-5.

[0066] After pMD-18T-Tum-5 was digested with SphI and HindIII, small fragments were recovered; plasmid pQE30 (product of Qiagen) was digested with SphI and BamH1, and large fragments were recovered. The above two fragments were ligated with T4 ligase, and the ligation product was transformed int...

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Abstract

The invention discloses a method for preparing for tumour formingblood vessel peculiarity binding polypeptide and the admixture protein of human-5. The tumour formingblood vessel peculiarity binding polypeptid-CNGRCVSGCAGRC can separately combine with the tumour formingblood vessel endothelium cell surface high effect expressed aminopeptidase N (CD13). It establishes tumour formingblood vessel peculiarity binding polypeptide and the admixture protein of human-5 and obtains the high effect express in bacillus coli. It also discloses a method for preparing for human Tum-5. The Tum-5 is formed by the 54-132 bits amino acid near the N end of the Tumstatin, which is the function structure area of the Tumstatin anti-vascularization.

Description

Technical field [0001] The invention belongs to gene cloning, gene recombination, expression of exogenous genes in prokaryotic cells, purification of target protein, in vitro activity determination and physicochemical determination of tumor neovascularization specific binding polypeptide and recombinant human Tum-5 fusion protein in the field of medical biotechnology Properties identification and other technologies relate to an anti-tumor drug and its preparation method, in particular to a tumor angiogenesis specific binding polypeptide and recombinant human Tum-5 fusion protein (Tum-5) that can specifically bind to tumor tissue angiogenesis endothelial cells and tumor cells. -5-NGR) and its preparation method. Background technique [0002] The application of the targeting peptide NGR has been introduced in more detail in the patent "Tumor angiogenesis specific binding polypeptide and human interferon alpha fusion protein and its preparation" (Patent No.: ZL 02139537.3), so I wil...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K19/00C12N15/09C12P21/02
Inventor 孟洁如马楠颜真张英起
Owner FOURTH MILITARY MEDICAL UNIVERSITY
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