Production and certification of an enzyme reference material for creatine kinase isoenzyme 2 (CRM 608)

F. Javier Gella, Elena Frey, Ferruccio Ceriotti, Amparo Galán, Anthony G. Hadjivassiliou, Mogens Hørder, Klaus Lorentz, Donald W. Moss, Françoise Schiele, Francesca Canalias*

*Kontaktforfatter for dette arbejde

Publikation: Bidrag til tidsskriftTidsskriftartikelForskningpeer review

Resumé

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.

OriginalsprogEngelsk
TidsskriftClinica Chimica Acta
Vol/bind276
Udgave nummer1
Sider (fra-til)35-52
Antal sider18
ISSN0009-8981
DOI
StatusUdgivet - 10. aug. 1998

Fingeraftryk

Certification
Creatine Kinase
Isoenzymes
Enzymes
Clinical Chemistry
Mercaptoethanol
Isoelectric Point
Enzyme activity
Molecular mass
Chromatography
Electrophoresis
Sodium Chloride
Serum Albumin
Adenosine Diphosphate
Quality control
Drying
Buffers
Ethanol
Calibration
Degradation

Citer dette

Gella, F. Javier ; Frey, Elena ; Ceriotti, Ferruccio ; Galán, Amparo ; Hadjivassiliou, Anthony G. ; Hørder, Mogens ; Lorentz, Klaus ; Moss, Donald W. ; Schiele, Françoise ; Canalias, Francesca. / Production and certification of an enzyme reference material for creatine kinase isoenzyme 2 (CRM 608). I: Clinica Chimica Acta. 1998 ; Bind 276, Nr. 1. s. 35-52.
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abstract = "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.",
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Gella, FJ, Frey, E, Ceriotti, F, Galán, A, Hadjivassiliou, AG, Hørder, M, Lorentz, K, Moss, DW, Schiele, F & Canalias, F 1998, 'Production and certification of an enzyme reference material for creatine kinase isoenzyme 2 (CRM 608)', Clinica Chimica Acta, bind 276, nr. 1, s. 35-52. https://doi.org/10.1016/S0009-8981(98)00097-7

Production and certification of an enzyme reference material for creatine kinase isoenzyme 2 (CRM 608). / Gella, F. Javier; Frey, Elena; Ceriotti, Ferruccio; Galán, Amparo; Hadjivassiliou, Anthony G.; Hørder, Mogens; Lorentz, Klaus; Moss, Donald W.; Schiele, Françoise; Canalias, Francesca.

I: Clinica Chimica Acta, Bind 276, Nr. 1, 10.08.1998, s. 35-52.

Publikation: Bidrag til tidsskriftTidsskriftartikelForskningpeer review

TY - JOUR

T1 - Production and certification of an enzyme reference material for creatine kinase isoenzyme 2 (CRM 608)

AU - Gella, F. Javier

AU - Frey, Elena

AU - Ceriotti, Ferruccio

AU - Galán, Amparo

AU - Hadjivassiliou, Anthony G.

AU - Hørder, Mogens

AU - Lorentz, Klaus

AU - Moss, Donald W.

AU - Schiele, Françoise

AU - Canalias, Francesca

PY - 1998/8/10

Y1 - 1998/8/10

N2 - 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.

AB - 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.

KW - Enzyme activity

KW - Reference material

KW - Standardisation

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U2 - 10.1016/S0009-8981(98)00097-7

DO - 10.1016/S0009-8981(98)00097-7

M3 - Journal article

C2 - 9760018

AN - SCOPUS:0031811908

VL - 276

SP - 35

EP - 52

JO - Clinica Chimica Acta

JF - Clinica Chimica Acta

SN - 0009-8981

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