Studies of a chromatographic method for assay of human follicle stimulating hormone

Abstract

In clinical laboratories the determination of Human Follicle Stimulating Hormone (hFSH) can aid the clinician in the diagnosis, treatment, and prognosis during endocrine malfunctions. Only a few years ago such determinations could not have been performed. The development of Radioimmunoassay (RIA) made it possible for the assay of hFSH with high specificity and sensitivity. Presently the clinically acceptable method for the assay of hFSH is RIA.

The object of this study was to design an alternative assay technique that can be compared to RIA. The test method did not determine the immunological activity but rather the hFSH total concentration. The hFSH was deparated from the other serum constituents by Sephadex gel filtration. The hFSH was shown to be in the ammonium bicarbonate eluent from the gel filtration by an RIA technique. A concentration step was performed prior to electrophoresis. Cellulose acetate was used for the support medium in the electrophoresis technique. The protein bands obtained through electrophoresis were stained by Ponceau S. Upon clearing the cellulose acetate plates scanning of the hFSH peaks were done by using a densitometer with an integration unit. This allowed quantitation of the hFSH. The integrator counts were related to the initial hFSH concentration. The test method values were then statistically compared to the comparative method which was RIA. The test method proved to have limited success in the determination of hFSH. The clinical acceptance of the alternative method will be dependent upon further testing using a larger number of samples.