Optimized preservation of isoforms of creatine kinase MM isoenzyme in plasma specimens and their rapid quantification by semi-automated chromatofocusing

D. R. Abendschein, H. L. Fontanet, R. Nohara

Research output: Contribution to journalArticlepeer-review

7 Scopus citations

Abstract

We report a convenient chromatofocusing procedure for rapid and sensitive quantification of isoforms of the MM isoenzyme of creatine kinase (EC 2.7.3.2) in plasma and efficient methods for preserving isoform profiles during handling of specimens. The assay involves use of prepacked, re-usable Mono P chromatofocusing columns and a 'Fast Protein Liquid Chromatograph' (FPLC) system with on-line detection of isoform enzymatic activity in column effluent. Profiles of isoforms are analyzed within 25 min with the use of a 1-mL column; the lower limit of sensitivity for CK activity is 5 mU, and recovery of each isoform is within 1% of the amount added to plasma. Collection of blood specimens in Vacutainer Tubes containing 28.5 μmol of EDTA (final concentration in plasma, 7 to 10 mmol/L) inhibited carboxypeptidase activity in plasma by 76%, sufficient to essentially abolish isoform conversion in vitro at room temperature. These methods should facilitate applications of isoform analysis for diagnosis of myocardial infarction and coronary artery recanalization.

Original languageEnglish
Pages (from-to)723-727
Number of pages5
JournalClinical chemistry
Volume36
Issue number5
DOIs
StatePublished - 1990

Keywords

  • coronary artery recanalization
  • heart disease
  • myocardial infarction
  • sample handling

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