Dynamic studies of antibody-antigen interactions with an electrolyte-gated organic transistor

Affinity-based biosensors employing surface-bound biomolecules for analyte detection are important tools in clinical diagnostics and drug development. In this context, electrolyte-gated organic transistors (EGOTs) are emerging as ultrasensitive label-free biosensors. In this study, we present an EGOT sensor integrated within a microfluidic system. The sensor utilizes the cytomegalovirus (CMV) phosphoprotein 65 as a biorecognition element to detect the pathological biomarker human anti-cytomegalovirus antibody in solution. The biorecognition element is grafted onto the gate electrode by exploiting the polyhistidine-tag technology. Real-time monitoring of the EGOT response, coupled with a two-compartment kinetic model analysis, enables the determination of analyte concentration, binding kinetics, and thermodynamics of the interaction. The analysis of the relevant kinetic parameters of the binding process yields a reliable value for the thermodynamic equilibrium constant and suggests that the measured deviations from the Langmuir binding model arise from the co-existence of binding sites with different affinities toward the antibodies.