Quantitative analysis for isoforms of creatine kinase MM in plasma by chromatofocusing, with on-line monitoring of enzyme activity

R. Nohara, B. E. Sobel, A. S. Jaffe, D. R. Abendschein

Research output: Contribution to journalArticle

12 Scopus citations

Abstract

Changes in the proportions of individual isoforms of the MM isoenzyme of creatinine kinase (CK; EC 2.7.3.2) in plasma promptly reflect both myocardial infarction and coronary recanalization. However, quantitative methods developed thus far are too slow or cumbersome for routine use in making clinical decisions. We report a convenient, quantitative chromatofocusing assay with on-line fluorometric detection of isoform activity in the column eluent that provides results within 40 min from the time of sample application. Sample eluted from a microbore chromatofocusing column (1.8-mL bed volume) is split between a reaction stream, into which CK reagents are added, and a reference stream. After incubation at 37°C, NADPH formed by reaction of isoforms with CK reagent is detected at 340 nm. The system can detect activity of individual isoforms in plasma samples having total CK activity ≥ 21 U/L (30°C). Results correlated closely with those obtained by previously validated, but slow, chromatofocusing (r = 0.98, n = 30) and protein immunoblotting (r = 0.90, n = 20) procedures.

Original languageEnglish (US)
Pages (from-to)235-239
Number of pages5
JournalClinical chemistry
Volume34
Issue number2
DOIs
StatePublished - Jan 1 1988

ASJC Scopus subject areas

  • Clinical Biochemistry
  • Biochemistry, medical

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