Bispecific antibody for hepatitis B surface protein, and use thereof

A bispecific antibody and surface protein technology, which is applied in the direction of antibodies, antiviral immunoglobulins, applications, etc., can solve problems such as limiting the development process, high development costs, and inability to meet clinical needs, achieving strong synergistic effects and low cost. Immunogenic effects

Active Publication Date: 2015-11-18
SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY
View PDF2 Cites 6 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] Due to the multifactorial nature of the disease, single-target therapy often cannot meet clinical needs [4]
There have been reports of preclinical experiments on the combined use of the two monoclonals in clinical trials [5] , but this combination of monoclonal antibodies h...

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Bispecific antibody for hepatitis B surface protein, and use thereof
  • Bispecific antibody for hepatitis B surface protein, and use thereof
  • Bispecific antibody for hepatitis B surface protein, and use thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Example 1: Bispecific Antibody A 3 B 5 - Preparation of BsAb

[0033] 1. Cloning of antibody light chain constant region and heavy chain constant region genes and construction of their vectors

[0034] Isolate healthy human lymphocytes with lymphocyte separation medium, extract total RNA with Trizol reagent, design primers according to the sequences reported in literature [8] and literature [9], and use QIAGEN OneStep RT-PCRKit to amplify the heavy chain and light chain constant regions of human antibodies Gene. The heavy chain and light chain constant region genes of the human antibody were respectively connected to the expression vector AbVec plasmid, and the antibody heavy chain constant region nucleotide vector IgG-AbVec plasmid and the antibody light chain constant region nucleotide vector Igκ-AbVec were respectively constructed Plasmid, after sequencing verification, it was confirmed that the correct clone was obtained. Wherein, the nucleotide sequence and ami...

Embodiment 2

[0044] Example 2, bispecific antibody A 3 B 5 -Specificity of BsAb

[0045] Detection of bispecific antibody A by Elisa method 3 B 5 - the ability of the BsAb to specifically bind the HBsAg protein.

[0046] The recombinant HBsAg protein and the control protein cIg were coated on the ELISA plate, and after being blocked by the blocking solution, the blank group, different concentrations of the control protein cIg, and different concentrations of expressed and purified A 3 D. 5 Antibody incubation, adding Goatanti-humankappa-HRP after washing the plate, TMB chromogenic solution for color development, microplate reader reads at 450nm wavelength.

[0047] Draw according to antibody concentration and corresponding OD value figure 1 ,Depend on figure 1The analysis shows that with the increase of the antibody concentration, the corresponding reading value of the HBsAg protein group is also correspondingly increased. However, the reading value of the coating control protein g...

Embodiment 3

[0048] Example 3, bispecific antibody A 3 B 5 - BsAb binding epitope identification

[0049] Combining the antigenic epitope region of hepatitis B surface antigen (HBsAg) with neutralizing activity reported in previous literature, a biotin-labeled short peptide of hepatitis B surface antigen was synthesized to measure the binding of fully human hepatitis B surface protein monoclonal antibody on HBsAg area. Such as figure 2 As shown in the HBsAg pattern diagram, the synthetic four biotin-labeled HBsAg short peptide P 1 (aa:104-120), P 2 (aa:121-137), P 3 (aa:139-148), P 4 (aa:149-163), P 1 -P 4 The amino acid sequence is shown in SEQ ID NO.9-SEQ ID NO.12. Among them, the antigenic short peptide P 1 and P 4 is a linear structure, P 2 and P 3 ring structure [10] .

[0050] Preliminary experiments showed that binding to the conformational epitope segment (P 2 and P 3 The monoclonal antibody neutralizing HBV activity is better than the linear epitope segment (P 1...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

PropertyMeasurementUnit
Molecular weightaaaaaaaaaa
Login to view more

Abstract

The invention provides a bispecific antibody A3B5-BsAb for a hepatitis B surface protein. The bispecific antibody is formed by combining an antibody A3D5 with an antibody B5H6. The bispecific antibody has better HBV virus neutralization ability and HBsAg release inhibition ability than single use of the antibody A3D5 and the antibody B5H6, and has strong synergistic effects, so the bispecific antibody is likely to prevent HBV infection related hepatitis, hepatic cirrhosis and liver cancer. The bispecific antibody is a completely humanized antibody obtained through cloning HBsAg specific memory B cells in the peripheral blood of hepatitis B vaccine inoculated volunteer, has lower immunogenicity than murine, chimeric and humanized antibodies, and can be used to prepare hepatitis B virus related hepatopathy prevention or treatment drugs or diagnose reagents.

Description

technical field [0001] The invention belongs to the field of biotechnology. More specifically, the invention discloses the preparation technology of anti-hepatitis B surface protein bispecific antibody and its application in preventing HBV infection and treating HBV-related hepatocellular carcinoma. Background technique [0002] Hepatitis B virus (HBV) is a double-stranded DNA virus, which can easily cause chronic hepatitis B after infection in the human body, and then lead to liver cirrhosis and (or) liver cancer. According to statistics, in recent years, there are about 350 million HBV-infected people in the world, and about 1 million to 1.5 million people die each year from acute or chronic HBV infection-induced liver failure, liver cirrhosis and / or liver cancer. Therefore, preventing HBV infection has become a worldwide public health problems [1] . Hepatocellular carcinoma (HCC, referred to as liver cancer) is closely related to viral hepatitis, is one of the most impo...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
IPC IPC(8): C07K16/08C12N15/13C12N15/63A61K39/395A61P31/20G01N33/569
Inventor 李博华孟艳春王华菁杨扬于晓杰
Owner SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap