DARSTELLUNG VON GONADOTROPIN AUS MENSCHLICHEN HYPOPHYSEN

Abstract

ABSTRACT Approximately 3000 human hypophyses were processed by a modification of the method of König & King (1950) in order to obtain potent gonadotrophic extracts. The final product, fraction E3, was found to have a total gonadotrophic activity of 200 HMG U./mg in the mouse uterus test and was 10 times as active as the FSH standard NIH-FSH-S 1 in the HCG augmentation test. MSH, ACTH and LTH contamination was insignificant. The specific activities obtained in other laboratories (Gemzell et al. 1958, 1960; Li 1960; Steelman et al. 1953, 1958, 1959) are not substantially higher. It may therefore be concluded that our relatively simple procedure is just as efficient as the more complicated, time consuming methods referred to above. Furthermore the overall recovery of our method seems to be higher. Attempts were made to obtain a further purification of fraction E3 by means of CMC, Sephadex and DEAE-Sephadex chromatography and also by means of flow-stream electrophoresis. DEAE Sephadex chromatography results in a slight increase in specific activity, which was, however, combined with a substantial overall recovery loss. The other fractionation methods employed gave no satisfactory results. A complete separation of FSH and ICSH activity even by means of CMC chromatography was not obtained. The total gonadotrophic activity per hypophysis was found to be approximately 1000 HMG units (average in unselected batches of hypophyses from various pathology institutes). Estimation in single hypophyses of men and women above the age of 45 resulted in higher values. Single estimations of total gonadotrophin activity in hypophyses of foetuses and of pregnant women have very low values, while in the hypophyses of children no gonadotrophic activity could be detected.