Kinetic analysis of the weak affinity interaction between tris and lysozyme

Abstract

a b s t r a c tThe biosensor based on total internal reflection imaging ellipsometry (TIRIE), regarded as an automotive real-time research approach for biomolecular interaction, is introduced to analyze the kinetic process of the weak interaction between tris and lysozyme. The experiment is performed by delivering lysozyme solution diluted to different concentrations to the biosensor substrate interface immobilized with tris. By applying pseudo-first-order interaction kinetics model, we are able to obtain the kinetic parameters from fitting experimental data. The calculated association rate constant and dissociation rate constant of tris and lysozyme interaction are in 10 À2 mol À1 s À1 and 10 3 s À1 magnitude, respectively. To further improve TIRIE!s ability for kinetically characterizing biomolecular interaction, a theoretical method to deduce associate rate constant before experiment is proposed.

Potential Modulation on Total Internal Reflection Ellipsometry

“…Since TIRE can obtain the precise refractive index and thickness of the formed layer, it is employed as a powerful tool to study in situ protein adsorption on the sensing surface, ,− low molecular weight environmental toxin registration, − genomic DNA adsorption and subsequent interactions, , disease diagnosis, , and weak affinity among biomolecules. , On the other hand, for many biochemical applications, aqueous electrolyte is used as a buffer. Because of the inner potential difference between the two phases, an electric double layer (EDL) would be formed at the interface.…”