Chromatography of adrenotropic hormone on ion-exchange columns.

Abstract

DURING the past year there have been several observations which have indicated that adrenotropic hormone (ACTH), as measured by the ascorbic acid depletion test1, might be a basic substance. Payne, Raben and Astwood2 observed that its activity could be adsorbed on powdered cellulose from dilute acetic acid solution and eluted again by 0.1 N hydrochloric acid, and these experiments agreed with results from preliminary trials in this laboratory on the behaviour of adrenotropic hormone when subjected to solvent partition (some of these initial experiments were carried out in collaboration with Dr. H. G. Khorana, of the Department of Chemistry, University of Cambridge) and preparative electrophoresis. Astwood, Raben, Payne and Grady3 have since utilized a single-step adsorption of adrenotropic hormone activity on oxycellulose (10.4 per cent COOH). We have concurrently been exploring its behaviour on columns of ‘Amberlite IEC-50’, which have previously been found to permit effective chromatographic purification of low-molecular weight basic proteins, such as ribonuclease4 and lysozyme5.