Purification of lutropin and follitropin in high yield from horse pituitary glands.
A method has been developed for the purification of equine lutropin (eLH) and equine follitropin (eFSH) from horse pituitary glands which attains high yields of both hormones in contrast to previous methods that were devoted to one or the other with inferior recovery of the hormones. Two-pass chromatography over CM-Sephadex was used to separate eLH from eFSH. Subsequent steps employing QAE (quaternary amino-ethyl)-Sephadex chromatography and gel filtration on Sephacryl S-200 produced highly purified hormone preparations. Yields of purified eLH and eFSH were 110 and 60 mg/kg of frozen pituitaries, respectively. Subunits were prepared by dissociation in 8 M guanidine HCl followed by either gel filtration (eLH) or gel filtration followed by QAE-Sephadex chromatography (eFSH). The hormones and their subunits were characterized by sodium dodecyl sulfate-gel electrophoresis, amino acid analysis, NH2-terminal analysis, and by LH and FSH radioligand receptor assays.
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D001681:Biological Assay; D002850:Chromatography, Gel / Q000379:methods; D002852:Chromatography, Ion Exchange / Q000379:methods; D000431:Ethanol; D005640:Follicle Stimulating Hormone / Q000302:isolation & purification*; D006736:Horses; D007986:Luteinizing Hormone / Q000302:isolation & purification*; D046911:Macromolecular Substances; D010902:Pituitary Gland / Q000032:analysis*
D000596:Amino Acids; D046911:Macromolecular Substances; D000431:Ethanol; D007986:Luteinizing Hormone; D005640:Follicle Stimulating Hormone