Bioluminescent assay of total and brain-specific creatine kinase activity in cerebrospinal fluid.
A bioluminescent assay based on the firefly luciferase reaction has been used for determination of creatine kinase activity in CSF. Activities as low as 0.1 U/L can be measured. The coefficient of variation at an activity level of 0.3-0.4 U/L was between 5 and 6%. The assay conditions optimized for serum specimens can be used for CSF. The adenylate kinase activity is almost completely inhibited, which simplifies the procedure. The creatine kinase/(CK) isoenzyme distribution was obtained using the bioluminescent assay in combination with immunoinhibition or ion exchange chromatography. All specimens contained both MM and BB activity, but no MB was found. The study indicates that the bioluminescent assay is useful in the determination of CK isoenzymes in CSF. The clinical importance of the observed CK levels will be reported in a separate communication.
D000244:Adenosine Diphosphate / Q000494:pharmacology; D000263:Adenylate Kinase / Q000134:cerebrospinal fluid; D001921:Brain / Q000201:enzymology; D003402:Creatine Kinase / Q000134:cerebrospinal fluid*; D004355:Drug Stability; D006801:Humans; D007527:Isoenzymes / Q000134:cerebrospinal fluid*; D008156:Luciferases; D008163:Luminescent Measurements; D008722:Methods; D013053:Spectrophotometry
D007527:Isoenzymes; D000244:Adenosine Diphosphate; D008156:Luciferases; D003402:Creatine Kinase; D000263:Adenylate Kinase