CVI Accepts, published online ahead of print on 18 March 2009

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Clin. Vaccine Immunol. doi:10.1128/CVI.00354-08
Copyright (c) 2009, American Society for Microbiology and/or the Listed Authors/Institutions. All Rights Reserved.

Development of a human-murine chimeric immunoglobulin M for use in the serological detection of human flavivirus antibodies

Brett A. Thibodeaux and John T. Roehrig^*

Arboviral Diseases Branch, Division of Vector-Borne Infectious Diseases, National Center for Zoonotic, Vector-Borne and Enteric Diseases, Coordinating Center for Infectious Diseases, Centers for Disease Control and Prevention, U.S. Department of Health and Human Services, 3150 Rampart Road, Fort Collins, CO 80521

^* To whom correspondence should be addressed. Email: jtr1{at}cdc.gov.

Current diagnosis of human flaviviral infections relies heavily on serological techniques such as the immunoglobulin (Ig) M antibody capture enzyme-linked immunosorbent assay (MAC-ELISA). Broad application of this assay is hindered by the lack of standardized positive human control sera that react with the wide variety of flaviviruses that can cause human disease, e.g., dengue (DEN), West Nile (WN), yellow fever (YF), Japanese encephalitis (JE), and Saint Louis encephalitis virus (SLEV). We have created a human-murine chimeric antibody combining the variable regions of a broadly flavivirus cross-reactive murine monoclonal antibody (MAb) 6B6C-1, and the constant region of human IgM to produce a standardized reagent capable of replacing human positive control sera in MAC-ELISA for diagnosis of all human flaviviral infections. Human-murine chimeric IgM antibody secreted from plasmid transformed Sp2/0-Ag14 cells had serological activity identical to 6B6C-1 as measured in ELISA, immunoblotting, and MAC-ELISA for multiple members of the flavivirus genus, including WNV, SLEV, YFV, DENV and JEV.