Purification and some properties of human urinary follicle stimulating hormone.

Abstract

A new approach to the preparation of biologically pure human urinary follicle stimulating hormone from human menopausal urine was performed. A crude human menopausal gonadotropin (HMG) with an activity of about 80-90 IU per mg was applied to a column filled with DEAE-cellulose, which had been previously equilibrated with 0.005M ammonium acetate buffer, pH 8.5. By a sodium chloride gradient' method on a variable gradient apparatus, the fraction of 0.08-0.12M sodium chloride concentration had 3 to 4 times higher in specific activity than the starting material. The active fraction was pooled, and was further purified by the gel filtration on Sephadex G-75 and the chromatography on CM-Sephadex C-25. This procedure yielded urinary FSH with an activity of more than 830 IU per mg.A homogeneous ultracentrifuge pattern was observed with this purified product, however, only faint-stained minor components still remained in the disc gel electrophoresis for further investigation. An isoelectric point of this. product estimated by the electrofocusing method was about pH 4.0. The LH/FSH ratio of our purified urinary FSH was less than 0.1.