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Traditional methods of monitoring biochemical reactions measure certain detectable reagents or products while assuming that the undetectable species follow the stoichiometry of the reactions. Here, based upon the metal-oxide thin-film transistor (TFT) biosensor, we develop a real-time molecular diffusion model to benchmark the concentration of the reagents and products. Using the nicotinamide adenine dinucleotide (NADH)-oxaloacetic acid with the enzyme of malate dehydrogenase as an example, mixtures of different reagent concentrations were characterized to extract the ratio of remaining concentrations between NAD + and NADH. We can thus obtain the apparent equilibrium constant of the reaction, (8.06 ± 0.61) × 10 4 . Because the whole analysis was conducted using a TFT sensor fabricated using a semiconductor process, our approach has the advantages of exploring biochemical reaction kinetics in a massively parallel manner.

acs omega

Analysis of the Biochemical Reaction Status by Real-Time Monitoring Molecular Diffusion Behaviors Using a Transistor Biosensor Integrated with a Microfluidic Channel