Effective purification of human chorionic gonadotropin and its subunits from pregnant women's urinary peptides absorbed on reverse-phase resin.
Human chorionic gonadotropin (hCG) was extracted and purified by reverse-phase resin (Sepralyte C8 resin) adsorption and Sephadex G-100 column chromatography from urine of pregnant women. Approximately 15000 IU of hCG was recovered from 1000 ml of urine by C8 resin adsorption. hCG from the gel-filtration step stimulated testosterone production in rat Leydig cells and had a specific activity of approximately 8000 IU/mg, a value which was higher than those of commercial hCGs. Furthermore, hCG purified to near-homogeneity was separated effectively into its subunits by reverse-phase high-performance liquid chromatography using an acetonitrile gradient in the presence of 0.1% trifluoroacetic acid with no special pretreatment for dissociation of subunits. The separated subunits were able to re-associate. The techniques used are simple and may be suitable for not only laboratory but also industrial production of hCG and its subunits.
Department of Biochemistry, School of Clinical Pharmaceutical Sciences, Nagasaki University, Japan.
R
R
Sakakibara
S
S
Miyazaki
T
T
Shigemura
N
N
Tominaga
A
A
Sakai
M
M
Ishiguro
D000818:Animals; D006063:Chorionic Gonadotropin / Q000302:isolation & purification* / Q000652:urine; D002852:Chromatography, Ion Exchange; D005260:Female; D006801:Humans; D008297:Male; D010455:Peptides / Q000652:urine*; D011247:Pregnancy; D051381:Rats; D012116:Resins, Plant; D013739:Testosterone / Q000032:analysis / Q000494:pharmacology
D006063:Chorionic Gonadotropin; D010455:Peptides; D012116:Resins, Plant; D013739:Testosterone