IMPROVED SEPARATION OF CREATINE-KINASE ISOENZYMES BY USE OF DEAE-SEPHAROSE CL-6B

  • 1 January 1980
    • journal article
    • research article
    • Vol. 26  (5) , 618-624
Abstract
A simple, rapid chromatographic procedure is described for quantitatively separating serum creatine kinase (CK) isoenzymes (EC 2.7.3.2), with DEAE-Sepharose CL-6B as the anion-exchanger. The column bed height and the elution parameters were established by a simplex procedure. DEAE-Sepharose CL-6B equilibrated in tris(hydroxymethyl)aminomethane (50 mmol/l, pH 7.5 and containing 30 mmol NaCl/l) is packed into a miniature polystyrene column with bed dimensions of 0.7 .times. 1.5 cm. The DEAE-Sepharose column system was evaluated and compared with a DEAE-Sephadex A-50 column system. The Sepharose column yields MM, MB and BB isoenzymes uniquely without cross contamination. Coefficients of variation (CV) for 10 replicate column assays were 2.8, 5.9 and 15.2% for 569 U MM/l, 82.3 U MB/l and 9.0 U BB/l, respectively. The serum sample was enriched with baboon heart extract. Day-to-day reproducibility for a serum control assayed on 10 days yielded CV of 4.6, 9.9 and 40.3% for 391, 45.3 and 1.9 U isoenzymes MM, MB and BB/l, respectively. A reference interval for each isoenzyme was derived from data on 81 men and 63 women. [CK-MB isoenzyme in serum is used in the diagnosis of acute myocardial infarction.].