Enzyme Immunoassay of 17 α-Hydroxyprogesterone with Microcentrifugal Analyzer Multistat III F/LS.

Abstract

we have applied an enzyme immunoassay for the determination of 17α-hydroxyprogesterone (17-OHP) in serum and dried blood sample on filfer paper to the“Multistat MCA III F/LS”Centrifugal Analyzer.
Glucose oxidase (GOD) was used as the label enzyme and conjugated with 17-OHP-3-(O-carboxymethyl) oxime.The immune reaction and separation of free from bound fractions were carried out in the sample cap coated with second antibody.The enzyme activity was measured by fluoresence reaction using Horseradish peroxidase and 3-(P-hydroxyphenyl) propionic acid after incubaticn with glucose.
The detection limit of H₂O₂, activity for GOD and 17-OHP are 5×10^-6M, 2×10^-6unit and 1 pg/tube, respectively.
The results were compared with those obtained by radioimmunoassay. Multistat MCA III F/LS is useful for the routine assay of enzyme immunoassay of 17-OHP.