Sensitive two-site sandwich enzyme immunoassay with antipeptide antibodies for detection of viral antigens.
Two-site sandwich ELISAs with antipeptide antibodies were developed for measuring the amounts of polyoma virus medium T and SV40 virus large T antigen in extracts from virus infected and transformed cells. In order to exclude crossreactive proteins, two antipeptide antibodies directed against different regions of the antigen were used in series. Both antigenic determinants have to be recognized for the assay to score positive. A positive signal was obtained with as little as 10 microliter of polyoma virus infected cell lysate and 100 microliter of extract prepared from medium T antigen-transformed FR18-1 cells which were shown to contain 37 and 15 pg of medium T antigen, respectively. This two-site sandwich ELISA is a sensitive tool to detect novel or closely related proteins.
Wiestler
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OD
Department of Pathology, University of California, San Diego, La Jolla 92093.
Walter
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G
D000818:Animals; D000914:Antibodies, Viral; D000952:Antigens, Polyomavirus Transforming / Q000032:analysis* / Q000276:immunology; D000956:Antigens, Viral / Q000032:analysis* / Q000276:immunology; D002461:Cell Line, Transformed; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D051379:Mice; D010455:Peptides / Q000276:immunology*; D013539:Simian virus 40 / Q000276:immunology
D000914:Antibodies, Viral; D000952:Antigens, Polyomavirus Transforming; D000956:Antigens, Viral; D010455:Peptides