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Clinical and Vaccine Immunology, May 2009, p. 679-685, Vol. 16, No. 5
1071-412X/09/$08.00+0     doi:10.1128/CVI.00354-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Development of a Human-Murine Chimeric Immunoglobulin M Antibody for Use in the Serological Detection of Human Flavivirus Antibodies{triangledown}

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, Colorado 80521

Received 26 September 2008/ Returned for modification 17 November 2008/ Accepted 5 March 2009

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

* Corresponding author. Mailing address: 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 Rd., Fort Collins, CO 80521. Phone: (970) 266-3541. Fax: (970) 266-3599. E-mail: epx1{at}cdc.gov

{triangledown} Published ahead of print on 18 March 2009.

Clinical and Vaccine Immunology, May 2009, p. 679-685, Vol. 16, No. 5
1071-412X/09/$08.00+0     doi:10.1128/CVI.00354-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





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