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Use of human antibody capable of neutralizing hepatitis b virus for the prevention or treatment of hepatitis b virus infection

A hepatitis B virus and human antibody technology, applied in the direction of antibodies, antiviral agents, medical preparations containing active ingredients, etc., can solve problems such as low specificity, non-therapeutic antibody source, infectious pathogen contamination, etc.

Inactive Publication Date: 2010-11-03
GREEN CROSS CORP THE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, currently available HBIG is not an ideal source of therapeutic antibodies due to its limited availability, low specificity and possible contamination with infectious pathogens

Method used

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  • Use of human antibody capable of neutralizing hepatitis b virus for the prevention or treatment of hepatitis b virus infection
  • Use of human antibody capable of neutralizing hepatitis b virus for the prevention or treatment of hepatitis b virus infection
  • Use of human antibody capable of neutralizing hepatitis b virus for the prevention or treatment of hepatitis b virus infection

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Example 1: Evaluation of the ability to neutralize HBV in chimpanzees

[0031] In order to confirm that the human antibodies of the present invention exhibit the ability to neutralize HBV in vivo, the following experiments were performed.

[0032] HBV 100 CID50 (50% chimpanzee infectious dose) obtained from Hepatitis Research Foundation (New York, USA) was put into three tubes. 0.1 mg and 10 mg of the antibody of the present invention (Hepabig-Gene, Green Cross, Korea) were added to two tubes, respectively, and no antibody was added to the other tube. PBS (Phosphate Buffered Saline) was added to the tube to a final volume of 3 ml, the mixture was incubated at 37°C for 1 hour, then at 4°C overnight, and frozen with liquid nitrogen to prepare test samples.

[0033] The test samples were administered intravenously to three chimpanzees (Hepatitis Research Foundation, New York, USA) who had not been previously infected with HBV. Antibody doses for each chimpanzee are lis...

Embodiment 2

[0048] Example 2: Evaluation of HBV Neutralization Ability in Mouse Model

[0049] 2-1) Construction of plasmid containing HBV DNA

[0050] The 1.3 sequence (Gene Bank accession No.DQ683578) of the HBV (adr subtype) gene (the HBV gene from the upstream of the enhancer I of the HBV genome to the downstream of the polyadenylation region; see figure 2 ) was inserted into the PmeI restriction site of pcDNA3.1 (Invitrogen, USA), and the resulting plasmid pHBV1.3-MBRI was prepared by using EndoFree PlasmidKit (Qiagen, Germany).

[0051] 2-2) Plasmid injection

[0052] 20 μg of the pHBV1.3-MBRI plasmid prepared in 2-1) was dissolved in physiological saline solution to a volume corresponding to 9% of the mouse weight, and injected into immunodeficiency at a rate of 0.3 ml / sec (hydrodynamic injection). In the tail vein of C57BL / 6J-Prkdcscid / SzJ female mice (8 weeks old, Jacksonlaboratory, USA).

[0053] 2-3) Administration of the antibody of the present invention and HBV replicat...

Embodiment 3

[0061] Example 3: Immunoprecipitation analysis of antibodies for HBV binding ability

[0062] Immunoprecipitation was performed to examine whether the antibody of the present invention binds to HBV in the blood of hepatitis B patients (provided by Ajou University, School of Medicine, Korea).

[0063] 3-1) Preparation of blood samples from patients with hepatitis B

[0064] 1000 μl of blood samples from patients with hepatitis B (diluted 10 times with 0.2% BSA / PBS buffer solution) were incubated with goat anti-human IgG (Fc-specific) agarose conjugate (Research Diagnostics Inc., Flanders, NJ) to remove immunoglobulins .

[0065] 3-2) Binding reaction between the antibody of the present invention and goat anti-human IgG agarose conjugate

[0066] 10 μl of antibodies of the invention (0.1, 0.5, 1 and 5 μg) and PBS were mixed with 50 μl of goat anti-human IgG agarose conjugate (Research Diagnostics) and incubated for 1 hour at room temperature with stirring. 10 mg of human im...

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Abstract

The present invention provides a use of a human antibody against a hepatitis B virus (HBV) surface antigen (HBsAg). The antibody exhibits an excellent HBV neutralizing ability, and thus, is useful for the prevention or treatment of the HBV infection or a disease caused thereby.

Description

technical field [0001] The present invention relates to the use of neutralizing HBV human antibody in preventing or treating HBV infection or diseases caused by it. Background technique [0002] Hepatitis B virus (HBV) causes hepatitis and liver cancer. WHO has revealed that about one-third of chronic HBV patients develop liver cirrhosis or liver cancer, and about one million people die from HBV-related diseases every year. [0003] The development of a vaccine against HBV has made it possible to prevent hepatitis B, but many people still suffer from chronic hepatitis through HBV infection. In addition, there is an increasing need for liver transplantation, which requires the development of effective antibodies that can inhibit HBV infection in liver transplantation. [0004] Viral replication inhibitors such as lamivudine are widely used as therapeutic drugs for chronic hepatitis B, but it is impossible to treat chronic hepatitis B using only viral replication inhibitors....

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K38/16A61K39/395
CPCA61K39/42A61K38/215A61K2039/505C07K16/082C07K2317/56A61P31/12A61P31/20A61K2300/00A61K38/16A61K39/395
Inventor 金世镐洪广元辛龙男辛庸源张基奂柳憬桓崔真设金判劲奉纪兑徐东赫吴宣姃
Owner GREEN CROSS CORP THE
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