HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

This Article Full Text Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via CrossRef Citing Articles via Google Scholar Google Scholar Articles by Artsaenko, O. Articles by Heintges, T. Search for Related Content PubMed PubMed Citation Articles by Artsaenko, O. Articles by Heintges, T. Agricola Articles by Artsaenko, O. Articles by Heintges, T.

Abrogation of hepatitis C virus NS3 helicase enzymatic activity by recombinant human antibodies

¹ Department of Gastroenterology, Hepatology and Infectious Diseases, Heinrich-Heine-University, Moorenstr. 5, 40225 Düsseldorf, Germany
² Department of Molecular Biotechnology, University of Aachen, Germany

Correspondence
Tobias Heintges
heintges{at}uni-duesseldorf.de

The hepatitis C virus (HCV) NS3 protein possesses both protease and helicase activities and is essential for virus replication and maturation. Specific inhibition of NS3 enzymatic activity can be achieved by antibody binding. Transduction of hepatocytes with encoding cDNA leading to intracellular expression of antibody fragments is expected to terminate HCV replication in infected cells. The objective of the present study was the generation of human antibody fragments that neutralize the viral NS3 helicase activity for gene therapeutic applications and drug design. A human immunoglobulin phage-display library cloned from bone marrow aspirate of patients infected with HCV was used for affinity selection against HCV NS3 helicase. Antibody fragments with high affinity to HCV helicase were isolated. To evaluate the inhibitory potential of isolated single-chain antibody fragments, a helicase-mediated, DNA-unwinding enzymatic assay was developed in ELISA format. Recombinant protein comprising the full-length HCV NS3 helicase domain was expressed in the baculovirus expression system. Recombinant antibodies that inhibit the HCV helicase at nanomolar concentrations, with efficacies ranging from 20 % to complete abrogation of enzymatic unwinding activity, were identified. These antibody fragments may be useful for novel gene therapeutic strategies that employ intracellular immunization and may provide new insights into the design of small molecule inhibitors of essential HCV proteins.

This article has been cited by other articles:

				B. HWANG, J. S. CHO, H. J. YEO, J.-H. KIM, K. M. CHUNG, K. HAN, S. K. JANG, and S.-W. LEE Isolation of specific and high-affinity RNA aptamers against NS3 helicase domain of hepatitis C virus RNA, August 1, 2004; 10(8): 1277 - 1290. [Abstract] [Full Text] [PDF]