Production and characterization of monoclonal antibodies against different epitopes of Ebola virus antigens - Monoclonal Antibodies

Production and characterization of monoclonal antibodies against different epitopes of Ebola virus antigens
Received 17 October 2006; revised 31 January 2007; accepted 5 February 2007. Available online 19 March 2007.
Soraya Shahhosseinia, 1, Dipankar Dasa, 1, Xiangguo Qiub, Heinz Feldmannb, Steven M. Jonesb and Mavanur R Suresha
Journal of Virological Methods
Volume 143, Issue 1, July 2007
ScienceDirect
Copyright ? 2007 Elsevier B.V. All rights reserved.
aFaculty of Pharmacy and Pharmaceutical Sciences, University of Alberta, Edmonton, AB, Canada T6G 2N8
bSpecial Pathogens Program, National Microbiology Laboratory, 1015 Arlington Street, Winnipeg, Manitoba R3E 3R2, Canada
Received 17 October 2006; revised 31 January 2007; accepted 5 February 2007. Available online 19 March 2007.
Abstract
Ebola virus (EBOV) causes hemorrhagic fever in humans and nonhuman primates with up to 90% mortality rate. In this study, Ebola virus like particles (EVLPs) and the aglycosyl subfragment of glycoprotein (GP1 subfragment D) were used to generate monoclonal antibodies (MAbs) against different epitopes of the viral antigens. Such MAbs could be useful in diagnostics and potential therapeutics of viral infection and its hemorrhagic symptoms. Hybridoma cell fusion technology was used for production of MAbs. The MAbs were characterized using ELISA and Western blot analysis. Furthermore, five recombinant sub-domains of GP1 subfragment D were produced, which were used as antigen in Western blot analysis for epitope mapping. Seventeen MAbs of different epitope specificities against EBOV antigens [virion protein (VP40), secreted glycoprotein (sGP), and GP1 subfragment D] were developed. Based on epitope mapping studies, the anti-GP MAbs were categorized into six groups. The binding of the three anti-sGP MAbs with different epitope specificities were mostly between aa 157 and 221. The two anti-VP40 MAbs with the same or overlapping epitopes are potentially good candidates for developing antigen detection assays for early diagnosis of EBOV infection. The anti-GP MAbs with different epitope specificities as an oligoclonal cocktail could be tested for therapy.
Keywords: Ebola virus; GP1,2; Monoclonal antibodies; Epitope mapping
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1 Equal contribution to this work
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