A New Recombinant Mouse-Human Antibody against Hepatitis-B
The technology suggests a process to generate a recombinant mouse-human chimeric antibody fragment, against Hepatitis B surface antigen. The process is consist 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).
Sector: Pharmaceuticals
Country: India
Area of Application: Pharma industry.
Keywords: hepatitis, antibody, Mouse-human chimeric antibody for hepatitis-B.
Advantages: Recombinant mouse-human chimeric Fab antibody. shows high affinity and specificity towards Hepatitis B surface antigen. Mouse-human chimeric antibody which can be safer and cheaper alternative. Have less antigenicity as compared to the mouse monoclonal antibodies. Would be more suitable for invivo use in humans. These are conducive to bio-neutralisation.
Environmental aspects: Not Applicable
Development Status: Laboratory Model
Legal Protection: Patent in Progress
Technical specifications:
Transfer Terms: Technology Licensing
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Country: India
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