Liquid-chromatographic separation and on-line bioluminescence detection of creatine kinase isoenzymes
Isoenzymes of creatine kinase were separated by anion-exchange chromatography, with use of an elution gradient containing lithium acetate (0.1 to 0.6 mol/L). A stream splitter was used to divert a 5% side stream of column effluent, which was subsequently mixed with the reagents necessary for bioluminescence assay of the separated isoenzymes. The use of the stream splitter greatly decreased the rate of consumption of reagent and, when combined with a peristaltic pumping system, permitted independent control of the side-stream flow rate. Thus both the residence interval in a delay coil in which the ATP reaction product is formed and the bioluminescence emission was monitored in a flow-through fluorometer without use of an external light source or filters. Separation and detection of the isoenzymes of creatine kinase were rapid, sensitive, and highly selective. The incremental decrease of bioluminescence response owing to inhibition by the ions in the eluent was less than 31% across the entire gradient.
- Research Organization:
- Oak Ridge National Lab., TN
- OSTI ID:
- 6639604
- Journal Information:
- Clin. Chem.; (United States), Vol. 26:6
- Country of Publication:
- United States
- Language:
- English
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Chromatographic separation and continuously referenced, on-line monitoring of creatine kinase isoenzymes by use of an immobilized-enzyme microreactor
Chromatographic separation and continuously referenced, on-line monitoring of creatine kinase isoenzymes by use of an immobilized-enzyme microreactor
Related Subjects
CREATINE
PHOSPHOTRANSFERASES
ISOENZYMES
BIOLUMINESCENCE
LIQUID COLUMN CHROMATOGRAPHY
ACETATES
ATP
DIAGRAMS
DOSE-RESPONSE RELATIONSHIPS
EXPERIMENTAL DATA
FLUORIMETERS
ION EXCHANGE CHROMATOGRAPHY
LITHIUM COMPOUNDS
LUCIFERASE
LUMINESCENCE
QUALITATIVE CHEMICAL ANALYSIS
QUANTITATIVE CHEMICAL ANALYSIS
SODIUM CHLORIDES
SODIUM COMPOUNDS
ALKALI METAL COMPOUNDS
AMINO ACIDS
CARBOXYLIC ACID SALTS
CARBOXYLIC ACIDS
CHEMICAL ANALYSIS
CHLORIDES
CHLORINE COMPOUNDS
CHROMATOGRAPHY
DATA
ENZYMES
HALIDES
HALOGEN COMPOUNDS
INFORMATION
MEASURING INSTRUMENTS
NUCLEOTIDES
NUMERICAL DATA
ORGANIC ACIDS
ORGANIC COMPOUNDS
OXIDOREDUCTASES
PHOSPHORUS-GROUP TRANSFERASES
SEPARATION PROCESSES
TRANSFERASES
550200* - Biochemistry