RAPID FLUORESCENCE FLOW-INJECTION IMMUNOASSAY USING A NOVEL PERFUSION CHROMATOGRAPHIC MATERIAL

A rapid fluorescence how injection immunoassay has been developed for a model analyte (testosterone) using a novel perfusion chromatographic matrix (POROS) coated with protein A. The fluorophore used was Texas Red. The perfusion matrix contains two classes of pores, which radically enhances mass transport and provides a very high capacity unaffected by how rate. For each assay the antibody-protein A reaction takes place at nearly neutral pH, and the complexes are eluted at acidic pH. Both preincubation and on-line assay formats were investigated. The total time for the pre-incubation and on-line assays was less than 4 and 6 min, respectively. Serum testosterone was determined by this method and good relative standard deviations and recoveries were achieved. The use of an avidin-biotin system in the preparation of the testosterone-Texas Red conjugate is also described.