Human Plasma β-Endorphin-Like Peptides: A Rapid, High Recovery Extraction Technique and Validation of Radioimmunoassay*

Abstract

This is a report of the development, calibration, and validation of a series of techniques required to measure beta-endorphin (beta-END)-like immunoreactivity in human plasma, including sieve and affinity chromatography. The RIA, which uses the antibody Brenda, is very sensitive (IC50 = 5-15 fmol/tube at a final concentration of 1:40,000). The extraction process, which uses the Sep-Pak C18 cartridge (Waters Associates, Inc.), is simple and rapid and has a recovery rate of more than 90%. It extracts proopiomelanocortin, beta-lipotropin, and beta-END. Physiological validation was provided by the measurement of beta-END-like immunoreactivity in a pool of plasma of normal humans (2.25 fmol/ml plasma), two pregnant women at term (9.5 and 10.75 fmol/ml), and one patient with Nelson's disease (2 pmol/ml plasma).