T. Tonai, K. Yokota, T. Yano
Oct 2, 1985
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0
Influential Citations
14
Citations
Quality indicators
Journal
Biochimica et Biophysica Acta
Abstract
Abstract A solid-phase enzyme immunoassay of 6-ketoprostaglandin F1α (a stable degradation product of prostaglandin I2) was developed in which the hapten molecule was labeled with β-galactosidase. The antiserum was bound to a polystyrene tube, and the enzyme-labeled and unlabeled 6-ketoprostaglandin F1/ga were allowed to react in a competitive manner with the immobilized antibody. The activity of the immunologically bound β-galactosidase was assayed by fluorometry, and correlated with the amount of unlabeled 6-ketoprostaglandin F1α According to a calibration curve 6-ketoprostaglandin F1α was detectable in a range of 30 fmol-10 pmol. When 6-ketoprostaglandin F1α was extracted from human serum by using an octadecylsilyl silica column (Sep-Pak C18) and the crude extract was subjected to the enzyme immunoassay, the content of 6-ketoprostagland F1α was 50–60 pmol / ml of human serum. An endogenous substance(s) which disturbed the immunoreaction and gave such an apparently high concentration of 6-ketoprostaglandin f1α was separated from the endogenous 6-ketoprostaglandin F1α by HPLC. With the purified 6-ketoprostaglandin F1α fraction there was a good correlation (r = 0.994) between enzyme immunoassay and radioimmunoassay. The validity of the enzyme immunoassay was confirmed by gas chromatography-selected ion monitoring.