Recombinant Mouse-Human Antibody against Hepatitis-B


 

Overview

The technology suggests a process to generate a recombinant mouse-human chimeric antibody fragment, against Hepatitis B surface antigen. The process consists of the below given steps:

1. Amplification of the variable region genes (VH and VL) of the anti-HBs mouse monoclonal 5S by reverse transcription (RT) followed by polymerase chain reaction (PCR). RNA isolated from 5S hybridoma was used as the source of these two genes.

2. Amplification of constant region of human kappa chain (Ck) and first constant region of human IgGl heavy chain (CH1) by reverse transcription (RT) followed by polymerase chain reaction (PCR). RNA isolated from human peripheral blood lymphocytes (PBLs) was used as the source of these two genes.

3. Mouse VL and human CK were linked by overlap PCR to generate chimeric light chain. Similarly mouse VH and CH1 were linked by overlap PCR to generate chimeric Fab.

4. Both of these chimeric antibody genes were cloned into a bicistronic bacterial expression vector (pCOMB3H, Scripps Research Institute, La Zola, USA) to express the chimeric Fab in the periplasm of E.coli (XLl-blue).

5. Recombinant clone was cultured in suitable medium and cell extract was prepared by the conventional method.

6. The recombinant chimeric antibody was purified by affinity chromatography using Protein G.

This recombinant molecule was generated by fusing variable region genes (Variable Heavy and Variable Low) of an anti-HBsAg mouse antibody 5S and constant region genes of human (CHl region of human IgGl and the CL region of human kappa chain).

The Problem Solved by the Technology

– Blood derived product is not manufactured in India. Anti-HBs HB1G is costly and can cause cross contamination
– Mouse antibodies are not suitable for therapeutic uses as they may generate immunogenic reaction in humans

How the Technology Solves the Problem

This recombinant molecule is generated by fusing variable region genes (Variable Heavy and Variable Low) of an anti-HBsAg mouse antibody 5S and constant region genes of human (CHl region of human IgGl and the CL region of human kappa chain).

Primary Application of the Technology

Antibody against Hepatitis B

Competitive Advantage

The technology suggests a process to generate a recombinant mouse-human chimeric antibody fragment, against Hepatitis B surface antigen

Notes on Development Status

Early stage

Patent application filed; available for licensing

Copyright © 2012 SkyQuest Technology Group: Building Invention Economy



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