Time-resolved immunofluorometric detection of antigens separated by high-performance liquid chromatography and coated to polystyrene.
We use high-performance liquid chromatography with fraction collection to separate an antigen of interest. The antigen is then immobilized on polystyrene microtiter wells and detected with a specific antibody, followed by a second biotinylated antibody and streptavidin labeled with a fluorescent europium chelate. Fluorescence can be quantified with the use of time-resolved fluorescence. Antigen detectability down to 2-3 x 10(-17) mol was achieved. This method could be used as an alternative to Western blot in certain applications.
Diamandis
E P
EP
Toronto Western Hospital.
Christopoulos
T K
TK
D000941:Antigens / Q000302:isolation & purification*; D002851:Chromatography, High Pressure Liquid; D005455:Fluorescent Antibody Technique; D007074:Immunoglobulin G / Q000302:isolation & purification; D000509:alpha-Fetoproteins / Q000302:isolation & purification
D000941:Antigens; D007074:Immunoglobulin G; D000509:alpha-Fetoproteins