Development of a human-murine chimeric immunoglobulin M antibody for use in the serological detection of human flavivirus antibodies.
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.
Thibodeaux
Brett A
BA
US Department of Health and Human Services, Division of Vector-Borne Infectious Diseases, Arboviral Diseases Branch, National Center for Zoonotic, Fort Collins, Colorado 80521, USA. epx1@cdc.gov
Roehrig
John T
JT
D000595:Amino Acid Sequence; D000818:Animals; D000914:Antibodies, Viral / Q000097:blood*; D001483:Base Sequence; D002460:Cell Line; D004797:Enzyme-Linked Immunosorbent Assay / Q000592:standards; D005416:Flavivirus / Q000276:immunology*; D018177:Flavivirus Infections / Q000175:diagnosis*; D006801:Humans; D007075:Immunoglobulin M / Q000235:genetics; D051379:Mice; D008969:Molecular Sequence Data; D011994:Recombinant Proteins / Q000235:genetics; D012015:Reference Standards; D017422:Sequence Analysis, DNA
D000914:Antibodies, Viral; D007075:Immunoglobulin M; D011994:Recombinant Proteins