Polymorphism of human pituitary FSH: analysis of immunoreactivity and in vitro bioactivity of different molecular species.
Follicle-stimulating hormone is known to be highly heterogeneous in serum and in the pituitary. In the present study, we have partially separated different molecular species of human pituitary FSH and characterized their immunoreactivity and in vitro bioactivity. Pooled extracts of male (n = 15) and female (n = 9) human pituitary glands were chromatographed on a column of Sephacryl S-200 and FSH-containing fractions were fractionated by chromatofocusing in the pH range 4-6. FSH was measured in the individual fractions by an in vitro bioassay, based on the FSH-dependent aromatase activity of immature rat Sertoli cells, and by the following methods based on commercial kits: radioimmunoassay (RIA), immunofluorimetric assay (IFMA), immunoradiometric assay (IRMA), immunoenzymometric assay (IEMA). In each assay, the kit standard, calibrated against the 2nd International Reference Preparation (IRP) 78/549, and the International Standard (IS) 83/575 were run in parallel. The relative potencies of the kit standards in terms of IS 83/575 were: IFMA 3.08, IRMA 1.62, RIA 2.42, IEMA 1.45 and bioassay 1.14. After chromatofocusing, pituitary FSH eluted mostly in fractions with pH approximately 4.5, without sex-related differences. In both sexes approximately 25% of bioactive material showed a pI < 4 and eluted with 1 M NaCl. Although the same IS 83/575 was used in the various assays, the profiles of immunoreactive FSH were significantly different. The highest intermethod variability was observed in the case of male pituitary FSH. The relative biopotency of the different molecular species of FSH did not appear to change according to their pI but, rather, varied with the assay method and the standard.(ABSTRACT TRUNCATED AT 250 WORDS)
Simoni
M
M
Institute of Reproductive Medicine of the University, Münster, Germany.
Jockenhövel
F
F
Nieschlag
E
E
D000368:Aged; D000369:Aged, 80 and over; D001681:Biological Assay; D002850:Chromatography, Gel; D005260:Female; D015200:Fluoroimmunoassay; D005640:Follicle Stimulating Hormone / Q000737:chemistry*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007124:Immunoenzyme Techniques; D015592:Immunoradiometric Assay; D008297:Male; D008875:Middle Aged; D010902:Pituitary Gland / Q000737:chemistry*; D011110:Polymorphism, Genetic; D011863:Radioimmunoassay; D012015:Reference Standards; D012737:Sex Factors
D005640:Follicle Stimulating Hormone