Impact of coenzyme regeneration on the performance of an enzyme-based optical biosensor: a computational study.
A mathematical model of a reagent-less optical sensing scheme composed of an enzymatic reaction coupled to light-controlled photochemical coenzyme regeneration is described. The model is based on previous experimental work describing the regeneration of NADPH from NADP(+) by excited state thionine coupled to the oxidation of isocitrate by isocitrate dehydrogenase. The system is capable of repeated isocitrate measurements without the addition of exogenous coenzyme. The model is simulated using numerical integration to determine the effect of regeneration on the sensor sensitivity, response time and maximum sample throughput rate. Prediction of these effects without a model is difficult due to activation and inhibition of the dehydrogenase by both forms of the coenzyme. The regeneration parameters, including thionine concentration and thionine excitation pattern, are varied to determine optimal sensor conditions to maximize performance. A periodic regeneration approach is found to be superior to a continuous regeneration approach as the former maximizes sensitivity and minimizes response time in most cases. In addition periodic regeneration results in a maximum sample throughput frequency that is achieved at a single optimal thionine level and is independent of the analyte concentration. In contrast the optimal thionine concentration during continuous regeneration varies with the sample analyte concentration. These findings highlight the importance of designing controllable regeneration for dehydrogenase-based optical biosensors.
Rickus
Jenna
J
Department of Agricultural and Biological Engineering, Purdue University, 225 S. University Street, West Lafayette, IN 47907, USA. rickus@purdue.edu
D015374:Biosensing Techniques / Q000295:instrumentation* / Q000379:methods; D003067:Coenzymes / Q000737:chemistry; D003198:Computer Simulation; D017076:Computer-Aided Design; D004789:Enzyme Activation; D004800:Enzymes, Immobilized / Q000737:chemistry; D004867:Equipment Design; D019544:Equipment Failure Analysis / Q000379:methods; D007521:Isocitrate Dehydrogenase / Q000737:chemistry*; D007523:Isocitrates / Q000032:analysis* / Q000737:chemistry*; D008956:Models, Chemical; D055095:Optics and Photonics / Q000295:instrumentation
D003067:Coenzymes; D004800:Enzymes, Immobilized; D007523:Isocitrates; C034219:isocitric acid; D007521:Isocitrate Dehydrogenase; C039355:isocitrate dehydrogenase (NADP+)