Bioluminescence enzyme immunoassay of 17-α-hydroxyprogesterone using glucose dehydrogenase as label

Abstract

A sensitive bioluminescence enzyme immunoassay for the determination of 17-α-hydroxyprogesterone (17-OHP) in dried blood samples spotted onto filter paper has been developed. 17-OHP was conjugated to glucose dehydrogenase by activated ester method. Separation of free and bound fractions after immune reaction was done by using polyacetal beads coated with purified second antibody. The enzyme activity was determined by using bacterial bioluminescence system. The sensitivity of the present method was 2.5 pg/assay tube for 17-OHP. The relative standard deviations of the intraand interassay were 3.9 % and 5.17.1 %, respectively.