METHOD OF OBTAINING AND PURIFYING HUMAN INHIBIN-A

摘要

FIELD: chemistry. ^ SUBSTANCE: disclosed is a method of obtaining and purifying human inhibin-A. Pieces of placenta are crushed and homogenised. A physiological solution is added in ratio of 1:2. Butyl alcohol is added to the obtained mass in ratio of 1:10 and then left for a day in a refrigerator at t +4C. The mixture is then centrifuged with diethyl ether in ratio of 1:3 for 10-12 minutes at 5000 rpm in a cold centrifuge twice. Supernatant fluid containing inhibin is put into a flask which is then put into a refrigerator at t +4C for a day. The supernatant fluid is centrifuged once more and over protein content therein is determined. The obtained solution is mixed with an equal volume of saturated ammonium sulphate solution and centrifuged in a cold centrifuge for 45-50 min at 10000 rpm. The obtained residue is dissolved in a tris-HCl buffer with pH 7.8, and then deposited on a lectin-sepharose column balanced with tris-HCl buffer. The entire volume of the dissolved residue obtained at the previous step is passed through. The column is washed with 1M sodium chloride solution buffered with tris-HCl buffer. The inhibin bound on the column is then eluted with 0.1M lactose solution in a borate buffer at pH 9.0. The eluate undergoes dialysis and concentration. ^ EFFECT: method enables to obtain inhibin-A with high output, high specific activity and high degree purification. ^ 1 tbl, 3 ex