Cel
Measurement of serum creatine kinase isoenzyme 2 (CK-MB) is
frequently requested from the clinical laboratory for the diagnosis
of acute myocardial infarction. A reference preparation of human
creatine kinase 2 could help to harmonise the results of
determinations carried out by different methods and to develop new
assay methods.
The objective of this work is the preparation of a batch of
approximately 2000 ampoules of lyophilized creatine kinase isoenzyme
2 originating from human heart tissue. The value of creatine kinase
catalytic concentration is assigned to the reference material by
means of a certification campaign.
The studies have been successfully completed and catalytic
concentration of creatine kinase 2 in samples of the reconstituted
material is certified to be 1.10 ñ 0.04 mkat/L or 66.0 ñ 2.4 U/L
(IFCC recommended method at 30 °C). The material can be used to rify
the comparability of results from different laboratories, for
intra-laboratory quality control, or for calibration of the creatine
kinase 2 catalytic concentration measurements. The preparations are
expected to be available at the end of 1997.
About 1.4 mg of creatine kinase 2 were purified from human heart by
ethanol precipitation and chromatography successively on
DEAE-Sephacel and Blue-Sepharose. The purified enzyme has a specific
activity of 17 mkat/mg protein 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
isoenzyme 2 had two subunits with a molar mass of 43650 (B) and
41700 (M) g/mol and an isoelectric pH of 5.8. The material was
prepared by diluting the purified creatine kinase 2 in a matrix
containing PIPES buffer 25 mmol/L, pH 7.2 ADP 2 mmol/L,
2-mercaptoethanol 5 mmol/L, sodium chloride 154 mmol/L and human
albumin 50 g/L. Samples of 1 mL were dispensed into ampoules and
freeze-drying.
Dziedzina nauki
Temat(-y)
Data not availableZaproszenie do składania wniosków
Data not availableSystem finansowania
CSC - Cost-sharing contractsKoordynator
8193 Barcelona
Hiszpania