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Time-resolved immunofluorometric detection of antigens separated by high-performance liquid chromatography and coated
E P Diamandis1, T K Christopoulos
1Toronto Western Hospital.
Biotechniques
|May 1, 1991
Abstract:
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.