INHIBIN ISOLATED FROM OVARIAN FOLLICULAR FLUID

摘要

A protein which satisfies all the biological criteriawhich are characteristic of inhibin has been isolated from agonadal source. The purification and characterization ofinhibin and the use of the purified material to raise antibodies,the use of inhibin and said antisera in a quantativeradioimmuno-assay, and applications in vitro and in vivo of inhibin andantibody to inhibin, are described.There is provided a purified protein, inhibin,characterised in that:a. the apparent molecular weight as determined by SDS-PAGEis 56,000 ~ 1,000b. the isoelectric point is in the range 6.9 - 7.3c. the protein can bind specifically to ConcanavalinA--Sepharosed. the protein consists of two sub-units, characterisedin that:i. their apparent molecular weights as determinedby SDS-PAGE are 44,000 ~ 3,000 and 14,000 ~2,000 respectively.ii. the isoelectric point of the 44,000 molecularweight sub-unit is in the range 6.0 - 7.0iii. the N-terminal amino acid sequences of thetwo sub-units are as described hereine. the protein can suppress follicle stimulating hormone(FSH) but not luteinising hormone (LH), thyroidstimulating hormone or prolactin in an in vitro bioassaysystemf. the protein can be labeled with radioactive iodine.There is also provided a method for isolating andpurifying inhibin from mammalian ovarian follicular fluid,characterised bya. one or more gel permeation chromatography steps;b. one or more reversed-phase high performance liquidchromatography steps;c. one or more preparative polyacrylamide gelelectrophoresis steps;d. Electrophoretic elution of the purified inhibin.