Characterizations of protein-ligand reaction kinetics by transistor-microfluidic integrated sensors
Journal
Analytica Chimica Acta
Journal Volume
1110
Pages
1-10
Date Issued
2020
Author(s)
Abstract
Understanding the binding affinities and kinetics of protein-ligand interactions using a label-free method is crucial for identifying therapeutic candidates in clinical diagnostics and drug development. In this work, the IGZO-TFT (thin-film transistor) biosensor integrated with a tailored microfluidic chip was developed to explore binding kinetics of protein-ligand biochemical interactions in the real-time manner. The IGZO-TFT sensor extracts the binding characteristics through sensing biomolecules by their electrical charges. Using lysozyme and tri-N-acetyl-D-glucosamine (NAG3) as an example, we established a procedure to obtain the parameters, such as the dissociation constant, Kd, and association rate constant, ka, that are critical to biochemical reactions. The correlation between the lysozyme concentration and TFT drain current signal was first constructed. Next, solutions of lysozyme and NAG3 of different mixing ratios were prepared. They were pre-mixed for various periods of reaction time before applying to the TFT sensor to extract signals of lysozyme molecules and the concentration remaining. With the knowledge of drain current changes at different reaction times, ka and Kd can be obtained. The values from our experiment are comparable to other methods, which suggests the proposed approach can be employed to explore protein-ligand interaction kinetics in the massively parallel manner if the TFT array is considered. ? 2020 Elsevier B.V.
Subjects
Association reactions; Binding energy; Biosensors; Dissociation; Drain current; Enzymes; Kinetics; Ligands; Microfluidics; Rate constants; Semiconducting indium compounds; Signal processing; Thin film circuits; Thin films; Association rate constants; Binding characteristics; Biochemical interactions; Dissociation constant; N-acetyl-D-glucosamine; Protein detection; Protein-ligand interactions; TFT (thin film transistor); Thin film transistors; ligand; lysozyme; n acetylglucosamine; protein; ligand; N,N',N''-triacetylchitotriose; trisaccharide; Article; association rate constant; binding affinity; chemical reaction kinetics; concentration (parameter); diffusion; dissociation constant; microfluidics; priority journal; protein interaction; chemistry; devices; genetic procedures; kinetics; metabolism; microfluidic analysis; transistor; Biosensing Techniques; Kinetics; Ligands; Microfluidic Analytical Techniques; Muramidase; Transistors, Electronic; Trisaccharides
Type
journal article