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Engineered polypeptide resisting infection of coated virus (hepatitis B virus) and its preparing process

A virus envelope and anti-virus technology, applied in the direction of viral peptides, anti-viral agents, anti-viral immunoglobulin, etc., can solve the problems of treatment failure and other problems, and achieve the effect of reducing toxic side effects and high targeting

Inactive Publication Date: 2003-02-12
CHENGDU PHOTON BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] However, in practical applications, the virus can quickly make the above treatments useless by taking advantage of its mutation characteristics
For example, the anti-hepatitis B virus drug Laminvudine, which was put into use several years ago, had an effective rate of almost 90% when it was first used. However, after only a few years of clinical application, due to the rapid mutation of the hepatitis B virus gene, the effective rate has dropped from the original 90%. down to only about 10%

Method used

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Embodiment 2

[0017] In this embodiment, the anti-HBV PreS1 scFv gene registered in Gene Bank AF427148 is linked to the carboxyl terminus of the colistin Ia channel domain to prepare an artificially combined anti-HBV engineering polypeptide plasmid, and the mutated plasmid is transfected into different In engineering bacteria containing plasmids, the bacteria were mass-proliferated (4L FB medium, 225rpm, 37°C; 6h), and the bacterial cells were centrifuged (4°C, 6000g, 20min). Take 4°C, 50mM boric acid buffer + 2mM EDTA+ 2mM DTT50-80ml suspension cells, ultrasonic disruption (4°C, 40W, 1-2min), high-speed centrifugation (4°C, 70000g, 1.5h), the supernatant was added to streptomycin sulfate to precipitate DAN, 4°C, 50mM boric acid buffer + 2mM EDTA + 2mM DTT 2L dialyzed overnight, loaded on CM ion exchange column, 0.1-0.3M NaCl + 50mM boric acid buffer gradient elution, the anti-HBV engineering with a purification rate of more than 90% was obtained Peptide, molecular weight about 40,000, prot...

Embodiment 3

[0019] In this embodiment, the anti-HBV PreS1 scFv gene registered in Gene Bank AF427148 is connected to the amino terminus of colistin Ia to prepare an artificially combined anti-HBV engineering polypeptide plasmid, and the mutated plasmid is transfected into the plasmid-free In the engineering bacteria, the bacteria were mass-proliferated (4L FB medium, 225rpm, 37°C; 6h), and the bacteria were centrifuged (4°C, 6000g, 20min). Take 4°C, 50mM boric acid buffer + 2mM EDTA + 2mM DTT 50 -80ml of suspended cells, sonicate (4°C, 40W, 1-2min), high-speed centrifuge to crush the cells (4°C, 70000g, 1.5h), take the supernatant and add streptomycin sulfate to precipitate DAN, 4°C, 50mM boric acid Buffer + 2mM EDTA + 2mM DTT 2L dialyzed overnight, loaded on the CM ion exchange column, 0.1-0.3M NaCl + 50mM boric acid buffer gradient elution, the anti-HBV engineering polypeptide with a purification rate of more than 90% was obtained, The molecular weight is about 90,000, and the protein c...

Embodiment 4

[0021]In this embodiment, the anti-HBV PreS1 scFv gene registered in Gene Bank AF427148 is connected to the carboxyl terminus of colistin Ia to prepare an artificially combined anti-HBV engineering polypeptide plasmid, and the mutated plasmid is transfected into the plasmid-free In the engineering bacteria, multiply the bacteria in large quantities (4L FB medium, 225rpm, 37°C; 6h), centrifuge and precipitate the bacteria (4°C, 6000g, 20min), take 4°C, 50mM boric acid buffer + 2mM EDTA + 2mM DTT 50 -80ml of suspended cells, sonicated (4°C, 40W, 1-2min), high-speed centrifugation (4°C, 70000g, 1.5h), supernatant was added to streptomycin sulfate to precipitate DAN, 4°C, 50mM boric acid Buffer + 2mM EDTA + 2mM DTT 2L dialyzed overnight, loaded on CM ion exchange column, 0.1-0.3M NaCl + 50mM boric acid buffer gradient elution, the anti-HBV engineering polypeptide with a purification rate of more than 90% was obtained, The molecular weight is about 90,000, and the protein content o...

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Abstract

An antivirus engineered polypeptide is composed of the polypeptide able to specifically bind coated virus antigen or receptor and the colicin able to form ion channel or its channel structure domain. It has two molecular structures divided according to different links between amino terminal and carboxyl terminal. Different engineered polypeptides for different viruses can be prepared by binding the colicin or its channel structure domain with the above-mentioned different polypeptide. Its advantage is sure effect on killing viruses.

Description

1. Technical field [0001] The present invention relates to an artificially combined antiviral engineering polypeptide and a preparation method thereof, by preparing an engineered polypeptide (Engineered Polypeptide) composed of different bacterial toxins and polypeptides that can specifically bind to viral envelope antigens or receptors, such as scFv domains peptide) to achieve antiviral purposes. 2. Background technology [0002] Viral infection has become a major threat to human life and health. As the earth's biosphere is increasingly destroyed by human industrialization, viral diseases, such as viral hepatitis, influenza, pneumonia, encephalitis, tumors caused by viruses, and AIDS, are becoming more and more rampant. Because the genes of viruses are prone to mutations, the damage caused by viruses is often involved in the malfunction of the host immune system. The mechanism is still being explored, so people have been working hard to try to prepare truly effective antivi...

Claims

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Application Information

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IPC IPC(8): A61K38/00A61P31/12C07K16/08C12N15/31
CPCC07K16/082C07K2317/622A61K38/00A61P31/12
Inventor 丘小庆
Owner CHENGDU PHOTON BIOTECH
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