Sensitive assay of creatine kinase isoenzymes in human serum using M subunit inhibiting antibody and firefly luciferase |
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Authors: | A Lundin I Styrélius |
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Institution: | 1. National Defence Research Institute, Department 4, S-172 04 Sundbyberg, Sweden;2. Karolinska Institutet at Huddinge Hospital, Department of Medicine, HuddingeSweden |
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Abstract: | A sensitive method for the assay of B subunit and total creatine kinase activity is described. The ATP formation in the creatine kinase reaction is continuously monitored by measuring the bioluminescence obtained with a purified firefly luciferase reagent. The B subunit activity is determined using an M subunit inhibiting antibody resulting in greater than 99.5% and approximately 50% inhibition of MM and MB isoenzymes, respectively. The bioluminescent method correlated well with a similar spectrophotometric method in the assay of B subunit as well as total creatine kinase activity (r greater than or equal to 0.98). However, the bioluminescent assay is considerably more sensitive, allowing the assay of B subunit activity in serum from healthy individuals. This is due to the inherent sensitivity of the bioluminescent assay of ATP, a reduced analytical interference from adenylate kinase and a reduced reagent blank. The within-series precision at 1 U/liter and greater than 52 U/liter corresponded to a C.V. of 14% and 3%, respectively. The method is as rapid and suitable for routine work as spectrophotometric methods. From a clinical point of view the new method is of particular interest in the early diagnosis of small acute myocardial infarctions. |
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Keywords: | CK creatine kinase (ATP:creatine phosphotransferase EC 2 7 3 2) AK adenylate kinase (ATP:AMP phosphotransferase EC 2 7 4 3) ALAT alanine aminotransferase (L-alanine:2-oxoglutarataminotransferase EC 2 6 1 2) DAPP inorganic pyrophosphate |
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