| 초록 |
We developed a label-free and washing-free method for the detection of creatine kinase (CK) activity based on a cascade enzymatic reaction (CER) and personal glucose meter (PGM). In this research, we newly designed a CER induced by the activities of CK and hexokinase (HK). In principle, when target CK present in the reaction solution, CK induces the phosphorylation of adenosine 5’-diphosphate (ADP) into adenosine 5’-triphosphate (ATP) by using creatine phosphate as a phosphate donor. Then, produced ATP can work as a phosphate source for the HK-mediated phosphorylation of glucose into glucose-6-phosphate, yielding ADP as a reaction product. Furthermore, produced ADP can participate in another cycle of enzymatic reaction, constituting a CER which eventually contributes to a significant reduction of glucose level in the reaction solution which can be easily measured by PGM. By this strategy, we successfully determined target CK activity down to 0.00147 U/mL with high specificity against other types of enzymes. Also, we reliably measured CK activity included in human blood, demonstrating the practical applicability of the strategy. |
