Production and certification of an enzyme reference material for creatine kinase isoenzyme 2 (CRM 608)
| Autor: | Donald W. Moss, Amparo Galán, Françoise Schiele, Elena Frey, Klaus Lorentz, Francesca Canalias, F. Javier Gella, Mogens Hørder, Ferruccio Ceriotti, Anthony G. Hadjivassiliou |
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| Rok vydání: | 1998 |
| Předmět: |
Sodium
Clinical Biochemistry chemistry.chemical_element Biochemistry Catalysis Reference Values Enzyme Stability medicine Humans Enzyme activity Polyacrylamide gel electrophoresis Creatine Kinase Ethanol precipitation chemistry.chemical_classification Chromatography biology Myocardium Biochemistry (medical) General Medicine Human serum albumin Chromatography Ion Exchange Enzyme assay Isoenzymes Kinetics Isoelectric point Enzyme Freeze Drying chemistry biology.protein Creatine kinase Electrophoresis Polyacrylamide Gel Reference material Standardisation medicine.drug |
| Zdroj: | Gella, F-J, Frey, E, Ceriotti, F, Galán, A, Hadjivassiliou, A G, Hørder, M, Lorentz, K, Moss, D W, Schiele, F & Canalias, F 1998, ' Production and certification of an enzyme reference material for creatine kinase isoenzyme 2 (CRM 608) ', Clinica Chimica Acta, vol. 276, no. 1, pp. 35-52 . https://doi.org/10.1016/S0009-8981(98)00097-7 |
| ISSN: | 0009-8981 |
| Popis: | We describe the preparation of a lyophilized material containing purified human creatine kinase 2 (CK-MB), and the certification of its catalytic concentration. The material can be used to verify the comparability of results from different laboratories, for intra-laboratory quality control, or for calibration of the creatine kinase 2 catalytic concentration measurements. The enzyme was purified from human heart by ethanol precipitation and chromatography successively on DEAE-Sephacel and Blue-Sepharose. The purified enzyme had a specific activity of 998.4 U/mg and was >99% pure on polyacrylamide gel electrophoresis. The material was examined for several possible contaminating enzymes, which were found to be absent. The purified creatine kinase 2 had two subunits (B and M) with molecular masses of 43 650 and 41 700 g/mol, respectively, and an isoelectric point at pH 5.8. The material was prepared by diluting the purified creatine kinase 2 in a matrix containing 25 mmol/L PIPES buffer, pH 7.2, 2 mmol/L ADP, 5 mmol/L 2-mercaptoethanol, 154 mmol/L sodium chloride and 50 g/L human serum albumin, dispensing it into vials and freeze-drying. The batch was shown to be homogeneous. The loss of enzyme activity on storage at -20°C is predicted to be less than 0.18% per annum on the basis of accelerated degradation studies. The catalytic concentration of creatine kinase in samples of the reconstituted material is certified to be 67.2±1.8 U/L (1.12±0.03 μkat/L) when measured, at 30°C, by the Recommended Method of the International Federation of Clinical Chemistry. Copyright (C) 1998 Elsevier Science B.V. |
| Databáze: | OpenAIRE |
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