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Exp Clin Endocrinol Diabetes 1986; 87(3): 277-287
DOI: 10.1055/s-0029-1210556
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© J. A. Barth Verlag in Georg Thieme Verlag KG Stuttgart · New York

Some Methodic Aspects in Optimizing the Radioimmunoassay of β-Endorphin

R. I. Rückert, Yvonne Hoffmann, W. Rohde
  • Institute of Experimental Endocrinology (Head: Prof. Dr. sc. med. G. Dörner), Humboldt University Medical School (Charité), Berlin/GDR
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Publication History

1985

Publication Date:
16 July 2009 (online)

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Summary

A specific double antibody radioimmunoassay (RIA) for human β-endorphin (β-EP) using an antibody to synthetic βh-endorphin was established. This antibody cross-reacted with β Lipotropin (β-LPH) at 42 per cent on molar basis and allowed a usable range of 20 pg to 4 ng of β-endorphin per ml in the assay.

Comparing the efficiency of the conventional extraction procedures under various conditions using Corning glass, Vycor glass, QUSO G-32, silicic acid and controlled pore glass 75, the optimal result was obtained by Corning glass, with a recovery rate of more than 80 per cent. The most simple and rapid method with an extraction efficiency of more than 90 per cent was found to be the extraction by use of Sep-Pak C18 cartridges.

The separation of β-endorphin from β-LPH was studied using Sephadex G-50, G-75, G-100 and Bio-Gel P-60 columns and different elution media. The use of a Sephadex G-50 column (0.9 X 55 cm) and elution with 0.1 N acetic acid-0.05 per cent HSA gave the best result.

The reliability of the radioimmunoassay was shown under physiological and pathophysiological conditions.

Key words

β-Endorphin - β-LPH - Radioimmunoassay - Insulin-hypoglycemia - Diseases of the hypothalamo-hypophysial-adrenal axis

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