Simultaneous assay of alpha-fetoprotein and free beta subunit of human chorionic gonadotropin by dual-label time-resolved immunofluorometric assay

Abstract

Abstract We developed a simple, rapid two-step dual-label assay for the noncompetitive determination of alpha-fetoprotein (AFP) and beta subunit of human chorionic gonadotropin (hCG beta) in serum. Monoclonal antibodies to detect AFP and hCG beta were labeled with europium (Eu) and samarium (Sm), respectively. Highly fluorescent chelates were developed by using the Delfia enhancement principle. The detection limits for AFP and hCG beta were approximately 0.02 kIU/L and approximately 0.2 IU/L, respectively. The within-run precision was < 5% over the whole range of AFP (1-500 kIU/L) and hCG beta (1-200 IU/L) concentrations tested. Cross-reaction of intact hCG was < 0.03%. The AFP concentrations determined with the dual-label assay correlated well with those obtained by Delfia AFP single-label kit. The concentrations of hCG beta were in good agreement with recently published data. Storing the serum samples for 24 h or 1 week at room temperature increased the hCG beta concentration by 4% and 26%, respectively. At 35 degrees C this dissociation of hCG increased 30-40-fold. Repeated freezing and thawing had no effect on the hCG beta concentration.