High affinity mouse-human chimeric Fab against Hepatitis B surface antigen

doi:10.3748/wjg.v11.i48.7569

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Dec 28, 2005 (publication date) through Apr 26, 2024

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World Journal of Gastroenterology

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1007-9327

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Baishideng Publishing Group Inc, 7041 Koll Center Parkway, Suite 160, Pleasanton, CA 94566, USA

Viral Hepatitis

World J Gastroenterol. Dec 28, 2005; 11(48): 7569-7578
Published online Dec 28, 2005. doi: 10.3748/wjg.v11.i48.7569

High affinity mouse-human chimeric Fab against Hepatitis B surface antigen

Biplab Bose, Navin Khanna, Subrat K Acharya, Subrata Sinha

Biplab Bose, Subrata Sinha, Department of Biochemistry, All India Institute of Medical Sciences, New Delhi, India

Navin Khanna, International Center for Genetic Engineering and Biotechnology, New Delhi, India

Subrat K Acharya, Department of Gastroenterology, All India Institute of Medical Sciences, New Delhi, India

Author contributions: All authors contributed equally to the work.

Supported by the Department of Biotechnology (DBT), Govt. of India, NO. BT/PR2540/PID/25/101/2001

Correspondence to: Professor Subrata Sinha, Department of Biochemistry, All India Institute of Medical Sciences, Ansari Nagar, New Delhi, PIN-110029, India. sub_sinha2004@yahoo.co.in

Telephone: +91-11-26594483 Fax: +91-11-26588663 / +91-11-26588641

Received: April 15, 2005
Revised: April 23, 2005
Accepted: April 26, 2005
Published online: December 28, 2005

Abstract

AIM: Passive immunotherapy using antibody against hepatitis B surface antigen (HBsAg) has been advocated in certain cases of Hepatitis B infection. We had earlier reported on the cloning and expression of a high affinity scFv derived from a mouse monoclonal (5S) against HBsAg. However this mouse antibody cannot be used for therapeutic purposes as it may elicit anti-mouse immune responses. Chimerization by replacing mouse constant domains with human ones can reduce the immunogenicity of this antibody.

METHODS: We cloned the V_H and V_L genes of this mouse antibody, and fused them with CH1 domain of human IgG1 and C_L domain of human kappa chain respectively. These chimeric genes were cloned into a phagemid vector. After initial screening using the phage display system, the chimeric Fab was expressed in soluble form in E. coli.

RESULTS: The chimeric Fab was purified from the bacterial periplasmic extract. We characterized the chimeric Fab using several in vitro techniques and it was observed that the chimeric molecule retained the high affinity and specificity of the original mouse monoclonal. This chimeric antibody fragment was further expressed in different strains of E. coli to increase the yield.

CONCLUSION: We have generated a mouse-human chimeric Fab against HBsAg without any significant loss in binding and epitope specificity. This chimeric Fab fragment can be further modified to generate a full-length chimeric antibody for therapeutic uses.

Keywords: Chimeric Fab, Hepatitis B surface antigen, Phage display