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Miniaturized polypeptide of anti EB Virus tumour, application and preparation method

A technology for Epstein-Barr virus and tumors, applied in the field of miniaturized anti-EB virus tumor polypeptides and its application and preparation, can solve the problems of large immunogenicity, large molecular weight, adverse allergies, etc., and achieve specific targeting effects

Inactive Publication Date: 2005-08-31
PROTEIN DESIGN LAB LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, because it is a whole colistin recombinant, its molecular weight is relatively large, and its immunogenicity may also be relatively large. Therefore, if patients use it for a long time, there is a hidden danger of causing adverse allergic reactions.

Method used

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  • Miniaturized polypeptide of anti EB Virus tumour, application and preparation method
  • Miniaturized polypeptide of anti EB Virus tumour, application and preparation method
  • Miniaturized polypeptide of anti EB Virus tumour, application and preparation method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Example 1 Construction of plasmids expressing anti-tumor polypeptides and preparation of recombinant miniaturized anti-EB virus tumor polypeptides

[0029] The original plasmid is the pET-15b commercial plasmid loaded with colistin Ia ion channel domain and Immunity protein gene (plasmid size 6.3kb, purchased from Novagen, the above-mentioned genes were loaded by our laboratory), and double-stranded oligonucleotide Point mutation technology (QuickChange TM Kit, Strategene Company) inserts the gene (the nucleotide sequence described in SEQID NO.3, 5, 7, 9, 11 and 13 in the sequence listing) of the coding guide polypeptide into the I626 site of the colistin Ia channel domain gene Finally, six recombinant plasmids pCHCEBCH1 to pCHCEBCH6 (such as Figure 1~6 shown). The recombinant plasmid was transfected into E.coli BL21(DE3)ply S engineering bacteria to prepare polypeptides, and six miniaturized anti-EB virus tumor polypeptides were obtained, the amino acid sequences o...

Embodiment 2

[0067] Example 2 Construction of plasmids expressing anti-tumor polypeptides and preparation of recombinant miniaturized anti-EB virus tumor polypeptides

[0068] The original plasmid was the pET-15b commercial plasmid (6.3 kb in size, purchased from Novagen, the above-mentioned gene was loaded by our laboratory) loaded with the colistin A ion channel domain and the Immunity protein gene. Point mutation technology (QuickChange TM Kit, Strategene Company) inserted the gene encoding the guide polypeptide (the nucleotide sequence described in SEQID NO.3 in the sequence listing) into the H592 site of the colicin A ion channel domain gene, and prepared the anti-tumor polypeptide A recombinant plasmid pCHCEBCH8 (such as Figure 8 shown). The recombinant plasmid was transfected into the E.coli BL21 (DE3) ply S engineering bacterium to prepare the polypeptide, and obtained the miniaturized anti-EB virus tumor polypeptide described in SEQ ID NO 30 in the sequence listing (referred t...

Embodiment 3

[0086] Example 3 Construction of plasmids expressing anti-tumor polypeptides and preparation of recombinant miniaturized anti-EB virus tumor polypeptides

[0087] The original plasmid is the pET-15b commercial plasmid loaded with the colistin Ia ion channel domain and the Immunity protein gene (the plasmid size is 6.3kb, purchased from Novagen, and the above-mentioned genes are loaded by our laboratory). Acid point mutation technology (QuickChange TM Kit, Strategene Company) inserted the gene encoding the guide polypeptide (the nucleotide sequence described in SEQ ID NO.3 in the sequence listing) before the D451 site of the colicin Ia ion channel domain gene, and prepared the anti-tumor polypeptide A recombinant plasmid pCHCEBCH7 (such as Figure 7 shown). The recombinant plasmid was transfected into E.coliBL21 (DE3) ply S engineering bacteria to prepare polypeptides, and obtained the miniaturized anti-EB virus tumor polypeptide described in SEQ ID NO 28 in the sequence lis...

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Abstract

A miniature polypeptide resisting to EB virus tumor, its gene and its recombinant plasmid are disclosed. Its preparing process includes linking the gene for coding and leading peptide with the plasmid carrying the colicin ion channel structure domain gene, transferring engineering bacteria for amplifying, and separating and purifying by His-tag column. It can be used to kill tumor cells.

Description

technical field [0001] The invention relates to an anti-EB virus tumor polypeptide gene, recombinant plasmid, polypeptide and its application and preparation method. Background technique [0002] Malignant tumors are a huge threat to human health. There are about 7 million patients who die from malignant tumors every year in the world, among which China accounts for one-sixth. Malignant tumors are the second cause of death in my country. Because its etiology, pathogenesis, and clinical manifestations have not yet been clarified, so the control effect is not ideal. Existing anti-tumor drugs play an important role in the treatment of tumors. Although they have achieved certain curative effects on some tumors, they still have defects such as poor selectivity to tumor cells, many and serious adverse reactions, and drug resistance. [0003] In recent years, great progress has been made in the basic and clinical research of tumors, and one of the most...

Claims

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Application Information

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IPC IPC(8): A61K47/48A61P35/00C07K14/435C07K16/08C12N15/12C12N15/63C12P21/02
CPCC07K16/085A61K47/48415C07K2319/00C07K2317/565A61K2039/505A61K47/48523A61K47/6811A61K47/6839A61P35/00
Inventor 丘小庆
Owner PROTEIN DESIGN LAB LTD
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