Rapid and sensitive quantitative immunoassay for the large simian virus 40 T antigen.
A quantitative, enzyme-linked immunoadsorbent assay has been developed for the simian virus 40 large T antigen. When hamster anti-simian virus 40 tumor serum was used, this method permitted specific identification of large T antigen and its analog, the D2 hybrid protein, a molecule with the same C-terminal approximately 600 amino acids as large T antigen. The sensitivity limit of this test was 0.63 ng of protein. The slopes of the regression lines of the enzyme-linked immunosorbent assay titrations performed with highly purified D2 or simian virus 40 large T antigen and with crude extracts of simian virus 40-infected monkey and transformed human cells were identical. Thus, the curve generated with a purified protein, such as D2, can serve as a quantitative standard for the measurement of large T antigen in a wide variety of extracts. Furthermore, solutions containing high salt concentrations and buffers containing up to 0.1% Nonidet P-40 did not interfere with the assay, making it applicable to the measurement of large T antigen in a variety of chromatographic fractions. The enzyme-linked immunosorbent assay was three times more sensitive, was significantly faster to perform, and was quantitatively valid over a much broader large-T-antigen concentration range than the complement fixation test. As such, it should be useful in future studies of the structure and function of this protein.
D000645:Ammonium Sulfate / Q000494:pharmacology; D000951:Antigens, Neoplasm / Q000032:analysis*; D000956:Antigens, Viral / Q000032:analysis*; D000957:Antigens, Viral, Tumor; D002460:Cell Line; D002471:Cell Transformation, Neoplastic; D002472:Cell Transformation, Viral; D003168:Complement Fixation Tests; D003902:Detergents / Q000494:pharmacology; D004797:Enzyme-Linked Immunosorbent Assay; D005990:Glycerol / Q000494:pharmacology; D017830:Octoxynol; D011092:Polyethylene Glycols; D013539:Simian virus 40 / Q000276:immunology*; D012965:Sodium Chloride / Q000494:pharmacology
D000951:Antigens, Neoplasm; D000956:Antigens, Viral; D000957:Antigens, Viral, Tumor; D003902:Detergents; D011092:Polyethylene Glycols; D012965:Sodium Chloride; D017830:Octoxynol; C010615:Nonidet P-40; D005990:Glycerol; D000645:Ammonium Sulfate