The preparation of bacterial luciferase conjugates for immunoassay and application to rubella antibody detection.
Bacterial luciferase has been modified with the thiolating reagent S-acetylmercaptosuccinic anhydride and covalently crosslinked to either Staphylococcus aureus protein A or anti-human immunoglobulin G (IgG) with the heterobifunctional reagent m-maleimidobenzoic acid N-hydroxysuccinimide ester. The conjugates retain enzymatic light-emitting activity and have the ability to bind IgG antibody. The ability of these conjugates to detect human IgG has been demonstrated by application to rubella immunity screening. Rubella antibodies are isolated from serum on the surface of rubella antigen-coated tubes and subsequently determined by light emitted from bound conjugate. In a preliminary study, the bioluminescent immunoassay has been compared to a commercial rubella antibody radioimmunoassay and found to be comparable in the ability to determine rubella immunity.
D000914:Antibodies, Viral / Q000032:analysis*; D003432:Cross-Linking Reagents; D006801:Humans; D007124:Immunoenzyme Techniques; D007074:Immunoglobulin G; D008156:Luciferases; D012409:Rubella / Q000276:immunology*; D013205:Staphylococcal Protein A; D013438:Sulfhydryl Compounds; D014733:Vibrio / Q000201:enzymology
D000914:Antibodies, Viral; D003432:Cross-Linking Reagents; D007074:Immunoglobulin G; D013205:Staphylococcal Protein A; D013438:Sulfhydryl Compounds; D008156:Luciferases