Three-dimensional epitope of hepatitis b surface antigen and antibody binding specifically thereto

A surface antigen, hepatitis B technology, applied in the direction of antibody medical components, virus antigen components, specific peptides, etc., to achieve the effect of eliminating hepatitis B virus

Pending Publication Date: 2021-10-29
GREEN CROSS CORP THE
View PDF4 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In addition, it has been reported that although the "a" determinant serves as the main neutralizing epitope for HBV, certain mutants with mutations within the "a" determinant that arise in some patients escape from hepatitis B vaccination antibody

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
  • Three-dimensional epitope of hepatitis b surface antigen and antibody binding specifically thereto
  • Three-dimensional epitope of hepatitis b surface antigen and antibody binding specifically thereto
  • Three-dimensional epitope of hepatitis b surface antigen and antibody binding specifically thereto

Examples

Experimental program
Comparison scheme
Effect test

preparation example 1

[0070] Preparation Example 1. Preparation of an antibody specifically binding to HBsAg

[0071] In the present invention, as the antibody specifically binding to HBsAg, a monoclonal antibody produced by a strain with accession number KCTC13760BP was used. In addition, for convenience, the monoclonal antibody produced by this strain was named GC-100A.

[0072] Experimental method 1. Native agarose gel electrophoresis (NAGE)

[0073] Huh-7 cells were transfected with empty (mock) vector or Flag-small HBsAg expression plasmid using Lipofectamine 3000. After 2 days, cells were lysed using RIPA buffer (Thermo Fisher, 89901). Centrifugation was performed at 4°C and 12,000 rpm for 15 minutes, and then the pellet-free supernatant was transferred to a new Eppendorf tube. To this was added 6X dye-loaded agarose (50% glycerol, 0.1% BPB) to a final IX concentration and mixed well. A 1.2% TBE agarose gel was prepared and the samples mixed with the dye-loaded agarose were loaded on it. ...

experiment example 1

[0087] Experimental example 1. Identification of the HBsAg epitope of GC-100A antibody

experiment example 11

[0088] Experimental Example 1.1. Identification of Possibility of Detection of HBsAg VLP Using GC-100A

[0089] Until 2016, ELISA was the only test that made it possible to detect HBsAg with GC-100A. In addition, it was not possible to detect HBsAg with GC-100A, which is considered to have a conformational epitope, in Western blot analysis using SDS-PAGE. Therefore, a technique for detecting HBV capsid in a native state was devised and an attempt was made to detect HBsAg VLP (consisting of 100 single HBsAg molecules) using NAGE disclosed in Experimental Method 1. Since NAGE is a method for identifying macromolecules without causing protein denaturation, this method makes it possible to detect HBsAg VLPs with an anti-Flag antibody with a linear epitope as well as with GC-100A with a conformational epitope ( figure 2 ).

[0090] There was a difference between the HBsAg VLP pattern detected by GC-100A and anti-Flag antibody. In each of the upper strips, the VLP can be said to...

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

No PUM Login to view more

Abstract

The present invention relates to a specific three-dimensional epitope of a hepatitis B surface antigen and a hepatitis B neutralizing antibody binding thereto. The epitope provided by the present invention has a specific three-dimensional structure. In addition, the three-dimensional epitope of the present application does not contain the 'a' determinant that may generate an escape mutation upon administration of conventional vaccines or HBIg. Thus, an antibody capable of binding to the three-dimensional epitope of the present application is highly unlikely to allow the emergence of a vaccine escape mutation, which is caused by conventional vaccines, and as such, can retain a sustained effect. Therefore, such an antibody or a vaccine composition can find effective applications in the prevention and treatment of HBV, having great economic value.

Description

technical field [0001] The present invention relates to the conformational epitope of hepatitis B surface antigen (abbreviated as HBsAg hereinafter) and the antibody specifically binding to it. Background technique [0002] Patients with chronic hepatitis B had no or very poor T cell responses compared with spontaneously healed patients. This has been reported to be because T cells specific for HBV disappear or fail to function properly due to continued exposure to excess viral antigens, such as hepatitis B surface antigen (HBsAg). [0003] For patients with chronic hepatitis B, viral particles in the blood reach 10 10 particles / ml, and HBsAg can even reach the level of 100ug / ml. Such excess viral particles or HBsAg suppresses the immune function of the human body, and thus is considered to be an important cause of chronic hepatitis. In addition, it has been reported that HBV particles or HBsAg can enter dendritic cells to reduce the activation of T cells, B cells and NK ...

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
Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/005C07K16/10A61K39/29A61P31/14
CPCA61P31/14C07K14/005C12N2730/10122C12N2730/10134C07K16/082C07K2317/34C07K2317/76A61K39/12A61K2039/575C07K16/109A61K39/29C12N2770/24222C12N2770/24234A61K39/292C12N2730/10022C12N2730/10034
Inventor 金政焕金佑眩
Owner GREEN CROSS CORP THE
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