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.
Sakakibara
R
R
Department of Biochemistry, School of Clinical Pharmaceutical Sciences, Nagasaki University, Japan.
Miyazaki
S
S
Shigemura
T
T
Tominaga
N
N
Sakai
A
A
Ishiguro
M
M
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