Clinical Chemistry
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Clinical Chemistry 33: 2000-2003, 1987;
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Clinical Chemistry, Vol 33, 2000-2003, Copyright © 1987 by American Association for Clinical Chemistry

Time-resolved immunofluorometric assay of alpha-fetoprotein in serum and amniotic fluid, with a novel detection system

MA Chan, AC Bellem and EP Diamandis
CyberFluor Inc., Toronto, Ontario, Canada.

We describe a new "sandwich"-type nonisotopic immunoassay of alpha- fetoprotein (AFP) in serum and amniotic fluid. In the assay, AFP binds to a monoclonal antibody immobilized in a microtiter well and to a polyclonal soluble biotinylated antibody. A fluorescent product is created on the solid-phase after adding streptavidin labeled with a new Eu3+ chelate, 4,7-bis(chlorosulfophenyl)-1,10-phenanthroline-2,9- dicarboxylic acid (BCPDA), and excess Eu3+. The fluorescent complex, monoclonal antibody-AFP-polyclonal antibody-biotin-streptavidin-BCPDA- Eu3+, is quantified by nitrogen laser excitation at 337.1 nm, the emission at 615 nm being monitored in an especially designed gated fluorometer working in a time-resolved mode. This assay of AFP has a broad dynamic range (up to 1 mg/L), and is precise and accurate. The detection limit is approximately 0.1 microgram/L. Results agree well with those obtained by established techniques.




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Copyright © 1987 by the American Association for Clinical Chemistry.