METHOD OF PREPARING VESICAL LIQUID α-GLOBULIN ASSOCIATED WITH TRUE PEMPHIGUS Russian patent published in 1995 - IPC

FIELD: medicine. SUBSTANCE: method of vesical liquid α₂-globulin preparing is developed. Method involves the precipitation of vesical liquid proteins with ammonium sulfate followed by its dissolving in 0.05M phosphate buffer, pH 7.4. Then preparation is purified by ion-exchange chromatography on DEAE-cellulose, and fraction eluted at 0.2-0.6M NaCl is separated. The latter is purified by hydrophobic chromatography on phenyl-Sepharose 4B, and fraction 0.6-0 M (NH₄)₂SO₄ is separated, followed by dialysis and gel-filtration on Sephadex G-200, and fraction with molecular mass 50-70 kD is collected, and affine chromatography on Con-A-agarose. The end product is a fraction eluting at the range of concentration of α-D-glycopyranoside from 2.5 to 3.0% its molecular mass is 60 ± 10 kD, relative electrophoretic mobility is 0.83 + 0.02, and isoelectric point pI 5.25 + 0.23. Based on prepared preparation method of high-quality antibody preparation can be developed. Antibodies to α₂-globulin of vesical liquid associated with true pemphigus can be used as diagnosticum of the true pemphigus and also for monitoring and therapy quality estimation. EFFECT: improved method of globulin preparing. 2 tbl