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

Mental Health Research Institute, University of Michigan, Ann Arbor, Michigan 48109

Address requests for reprints to: Dr. Cheryl A. Cahill, Mental Health Research Institute, University of Michigan, 205 Washtenaw Place, Ann Arbor, Michigan 48109.

This is a report of the development, calibration, and validation of a series of techniques required to measure β-endorphin (β-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, β-lipotropin, and β-END. Physiological validation was provided by the measurement of β-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).(J Clin Endocrinol Metab 56: 992, 1983)

^* This work was supported by the following grants: NIDA Grant DA-02265 (to H.A.), NIDA Center Grant DA-00154 (to H.A.), NIMH Grant ROl-MH-36168 (to H.A.), National Research Service Award Fellowship 1F31-NU-05337 (to C.A.C.), and American Nurses' Foundation, Ethicon Scholar (to C.A.C.).

Received May 3, 1982.

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