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Humanized single chain antibody of anti-hepatitis B virus surface antigen a determinant and application thereof

A surface antigen, single-chain antibody technology, applied in antiviral agents, antibodies, digestive systems, etc., can solve problems such as application value limitations

Inactive Publication Date: 2010-03-03
HANGZHOU IMMUSOURCE BIOTECHN
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the hepatitis B virus immunoglobulin currently used clinically is isolated from the serum of normal people who have received hepatitis B vaccination and belongs to blood products, so it has the potential risk of spreading some blood-borne infectious diseases, such as AIDS, Hepatitis C, so its application value is greatly limited

Method used

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  • Humanized single chain antibody of anti-hepatitis B virus surface antigen a determinant and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0017] (1) Isolation of lymphocytes

[0018] A total of 100ml of peripheral blood was collected from 3 normal persons with high titers of protective antibodies who received hepatitis B vaccination, added an equal volume of lymphocyte separation medium, and centrifuged for stratification at 2500g for 10min. Carefully absorb the middle lymphocytes and centrifuge at 2500g for 10min. Discard the supernatant, and use 1ml Trizol / 5ml peripheral blood to lyse the lymphocytes until the solution is completely transparent; obtain the lymphocyte lysate.

[0019] (2) Extraction of RNA

[0020] Incubate the lymphocyte lysate at room temperature for 5min, add 0.2ml chloroform / ml Trizol, shake for 15s with Votex, let stand at room temperature for 3min, 12000g, 5min. Take the upper clear aqueous phase, add 0.5ml isopropanol / ml Trizol, mix by inversion, precipitate at -20°C for 30-60min, and centrifuge at 12000g for 10min. Keep the precipitate, wash it once with 70% ethanol, air dry, and dis...

Embodiment 2

[0088] The single-chain antibody of the present invention is identified according to the above-mentioned method, and the results are as follows: figure 1 . according to figure 1 , we draw the following conclusions: the affinity between the single chain antibody of the present invention and the determinant of hepatitis B surface antigen a is as high as Kd=2nM, and has the potential function of neutralizing hepatitis B virus.

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Abstract

The invention discloses a humanized single chain antibody of anti-hepatitis B virus surface antigen a determinant, the amino acid sequence thereof is SEQ ID NO: 1. The VH amino acid sequence of the antibody is SEQ ID NO: 2, and the VL amino acid sequence of the antibody is SEQ ID NO: 3. The humanized single chain antibody of the anti-hepatitis B virus surface antigen a determinant is used for preparing preventive passive immunity drug or preparing medicines for treating chronic viral hepatitis B.

Description

technical field [0001] The invention belongs to genetic engineering antibodies, and relates to a humanized single-chain antibody against the surface antigen a determinant of hepatitis B virus and its application in the development of preventive passive immunization and potential therapeutic drugs for chronic hepatitis B virus. Background technique [0002] Viral hepatitis B is a worldwide disease caused by the hepatitis B virus (HBV). The incidence rate in developing countries is very high. According to statistics, there are more than 300 million asymptomatic hepatitis B virus carriers (HBsAg carriers) in the world, and my country accounts for about 93 million. One-third of hepatitis B virus carriers will turn into chronic hepatitis, 10% of patients with chronic hepatitis will develop liver cirrhosis, and 10% of patients with liver cirrhosis will further develop into liver cancer. Thereby hepatitis B virus is a kind of infectious disease which is very popular, extremely har...

Claims

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Application Information

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IPC IPC(8): A61K39/395A61P31/20A61P1/16
Inventor 韩森韩月恒刘向昕
Owner HANGZHOU IMMUSOURCE BIOTECHN
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