Separation of tissue and serum creatine kinase isoenzymes by ion exchange column chromatography

Abstract

The author describes a simple, rapid anion exchange column chromatographic technique for separating the creatine kinase (CK) isoenzymes in human serum and tissue. Extracts of CK rich tissues (skeletal muscle, cardiac muscle, and brain) were used to determine optimum conditions for separating CK isoenzymes MM, MB, and BB. Samples layered on mini columns (0.5 x 6.0 cm) of DEAE Sephadex A 50, were eluted stepwise with Tris buffered sodium chloride (100, 200, and 300 mmol/liter). Column effluents were assayed by the Rosalki CK method. Distribution of total activity among the eluted fractions was tissue specific and reproducible. Evaluation of sera from 71 patients with myocardial infarction and other diseases associated with elevated CK activity revealed isoenzyme patterns that resembled those of either cardiac muscle or skeletal muscle. Cardiac pattern (presence of MB isoenzyme) and clinical documentation of myocardial infarction were 100% correlated in the 35 patients so studied.