PURIFICATION AND PROPERTIES OF CHORIONIC GONADOTROPHIN FROM TROPHOBLASTIC TISSUE, URINE AND PLASMA OF A PATIENT WITH A HYDATIDIFORM MOLE

Abstract

SUMMARY Purified human chorionic gonadotrophin (HCG) fractions were obtained from urine, peripheral plasma and chorionic tissue of a woman with hydatidiform mole by a combination of DEAE-cellulose and DEAE-Sephadex ion exchange chromatography and gel filtration on Sephadex G-100. Homogeneity of the purified fractions obtained was demonstrated by analytical polyacrylamide gel electrophoresis and immunoelectrophoresis. The preparation from plasma had a higher ratio of biological to immunological activity than the urine or tissue preparation. The biological potency of the plasma HCG was 23000 i.u./mg, whereas the urinary and the trophoblastic tissue preparations showed a potency of 20000 and 6500 i.u./mg, respectively. Further characterization of the three fractions was achieved by analytical isoelectric focusing and determination of the molecular weight. The peak of radioimmunological activity after isoelectric focusing was found to be, in all three cases, at pH 4·90 and the molecular weight obtained after incubation with sodium dodecyl sulphate and urea was around 63000 When the molecular weight was determined after incubation in a medium containing 4 m-urea, sodium dodecyl sulphate and 2-mercaptoethanol two separate fractions were obtained from each of the three preparations. The heavier fraction showed a molecular weight of approximately 35000 and the lighter fraction had a molecular weight of approximately 27000.