| Research Abstract |
Immunoaffinity chromatography is by far the most useful and successfully applied method for purification of the inhibin in follicular fluid. Therefore, we tried to develop the immunoaffinity chromatography with polyclonal antibody to 32-KDa bovine inhibin purified by the improved methods on large scale. Human inhibins were purified from 10 liter human follicular fluid by various kinds of chromatography including Matrex gel red A, immunoaffinity chromatography, gel chromatography and revesed-phase HPLC.
On SDS-PAGE under nonreducing conditions, bioactive inhibin protein had two band migrating at MW 34,000 and 29,000 and similarly, immunoreactive inhibin protein had two bands migrating at MW 30,000 and 26,000, respectively.
cDNA clone encoding human inhibin alpha-and betaB subunits were isolated from a human testicular cDNA library. The cDNAs were linked to an expression vector and transfected into Chinease hamsteer cells (CHO cells). The cells transfected with the betaB subunit cDNA alone and with both the alpha and betaB subunit cDNAs secreted betaB dimer and inhibin B and betaB homodimer could evoke their biological actions on the FSH secretion from rat pituitary cultured cells.
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