PROCESS FOR MANUFACTURE OF COMPLEX INCLUDING SUPEROXIDE DIS-MUTASE AND CATALASE FROM HUMAN BLOOD ERYTHROCYTES Russian patent published in 1995 - IPC

FIELD: medical biochemistry, in particular, enzymology. SUBSTANCE: essence of this method resides in washing out human blood erythrocytes using physiologic salt solution, centrifuging these at 2,500 g during 15 min, lysing erythrocytes with equal amount of distilled water at temperature of -4 C during four hours, adding 0.3 to 0.5 percent by volume of urea to lysate, treating lysate of erythrocytes with admixture of chloroform and ethanol, ratio between lysate, chloroform and ethanol being 1:0.15:0.25, during 40 to 60 min, centrifuging stock, removing precipitate, adding 0.1 normal solution of hydrochloric acid (pH value: 3.7 to 4.2) to supernatant liquid, standing precipitate during 10 to 12 h, repeatedly centrifuging resultant product, adding acetone to thus prepared supernatant liquid (ratio: 1: 1) at temperature of -4 C, repeatedly centrifuging stock, dissolving precipitate in phosphate buffer solution having pH value of 7.4 to 7.8, carrying out dialysis against phosphate buffer, and finally carrying out sterilizing filtration of desired product. End product contains one fraction of catalase and two fractions of superoxide dis-mutase. Electrophoretic purity of end product equals to 99 % and contents of superoxide dis-mutase, catalase and impurities amount to 33.3 %, 66.6 % and 0.1 %, respectively. EFFECT: disclosed method allows reducing time needed for manufacturing end product as compared with prior art (50 to 56 h against 26 to 36 h) and intensifying processing.