Development of human-murine chimeric immunoglobulin G for use in the serological detection of human flavivirus and alphavirus antibodies.
Diagnosis of human arboviral infections relies heavily on serological techniques such as the immunoglobulin M (IgM) antibody capture enzyme-linked immunosorbent assay (MAC-ELISA) and the indirect IgG ELISA. Broad application of these assays is hindered by the lack of standardized positive human control sera that react with a wide variety of flaviviruses (e.g., dengue, West Nile, yellow fever, Japanese encephalitis, Saint Louis encephalitis, and Powassan viruses), or alphaviruses (e.g., Eastern equine encephalitis, Western equine encephalitis, Venezuelan equine encephalitis, and chikungunya viruses) that can cause human disease. We have created human-murine chimeric monoclonal antibodies (cMAbs) by combining the variable regions of flavivirus (6B6C-1) or alphavirus (1A4B-6) broadly cross-reactive murine MAbs (mMAbs) with the constant region of human IgG1. These cMAbs may be used as standardized reagents capable of replacing human infection-immune-positive control sera in indirect IgG ELISA for diagnosis of all human flaviviral or alphaviral infections. The IgG cMAbs secreted from plasmid-transformed Sp2/0-Ag14 cells had serological activity identical to that of the parent mMAbs, as measured by ELISA using multiple flaviviruses or alphaviruses.
Thibodeaux
Brett A
BA
Arboviral Diseases Branch, Division of Vector-Borne Diseases, National Center for Zoonotic Vector-Borne and Enteric Diseases, Centers for Disease Control and Prevention/U.S. DHHS, 3150 Rampart Road, Fort Collins, CO 80521, USA. epx1@cdc.gov
Panella
Amanda N
AN
Roehrig
John T
JT
D000524:Alphavirus / Q000276:immunology; D018354:Alphavirus Infections / Q000175:diagnosis*; D000818:Animals; D000911:Antibodies, Monoclonal / Q000235:genetics; D000914:Antibodies, Viral / Q000097:blood*; D005416:Flavivirus / Q000276:immunology; D018177:Flavivirus Infections / Q000175:diagnosis*; D006801:Humans; D007118:Immunoassay / Q000379:methods / Q000592:standards; D007074:Immunoglobulin G / Q000235:genetics; D051379:Mice; D008969:Molecular Sequence Data; D011994:Recombinant Proteins / Q000235:genetics; D012015:Reference Standards; D017422:Sequence Analysis, DNA; D014773:Virology / Q000379:methods* / Q000592:standards*
D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D007074:Immunoglobulin G; D011994:Recombinant Proteins