Development of a Microtiter Plate Enzyme-linked Immunosorbent Assay for 17 α, 20 β-21-trihydroxy-4-pregnen-3-one, a Teleost Gondadal Steroid

Abstract

A simple solid-phase microtiter plate enzyme-linked immunosorbent assay (ELISA) for 17 α, 20 β, 21-trihydroxy-4-pregnen-3-one (20 β-S), a teleost maturation inducing steroid, was developed using 20 β-S-3-CMO-seradish peroxidase as the label, and a homologous antiserum raised in rabbits. Second antibody coated to the plate was used to capture the antiserum with labeled or unlabeled 20 β-S. Within three hours, a steep standard curve covering 1.5-192 pg/well was obtained. The sensitivity and precision of this ELISA was almost the same as those of ordinary steroid adioimmunoassays. This non-isotopic assay system also allows prompt measurement of 20 β-S in catfish plasma with good specificity.