Quantitative determination of parathyroid hypertensive factor by enzyme-linked immunosorbent assay

Abstract

A new competitive enzyme immunoassay for the detection parathyroid hypertensive factor (PHF) in human plasma using a PHF–horseradish peroxidase conjugate and IgM antibody adsorbed on the microtiter plate was established. The antibodies raised against rat PHF could recognize human PHF. Cross-reactivity of anti-PHF antibodies with other serum haptens and proteins was negligible. Conjugation of PHF with horseradish peroxidase did not neutralize the antigen activity. The limit of detection of PHF was 0.02 U/mL in reference units and PHF levels between 0.02 and 1 U/mL could be detected. Within-run coefficient of variation (CV) was less than 10%, and between-run CV was less than 15% for over the dynamic range of the assay. Preliminary clinical studies were performed with plasma samples from hypertensive patients with confirmed diagnosis. Parathyroid hypertensive factor levels, as detected with this immunoassay, were positively correlated with PHF levels detected with the semiquantitative blood pressure (BP) bioassay previously used. Parathyroid hypertensive factor levels detected with the enzyme-linked immunosorbent assay (ELISA) were also correlated with BP in patients. The PHF ELISA provides a selective, simple, and rapid method that can be used for routine determination of PHF in human plasma, and provides useful clinical information.