Nonlinear analyte concentration gradients for one-step kinetic analysis employing optical microring resonators.
Conventional methods to probe the binding kinetics of macromolecules at biosensor surfaces employ a stepwise titration of analyte concentrations and measure the association and dissociation to the immobilized ligand at each concentration level. It has previously been shown that kinetic rates can be measured in a single step by monitoring binding as the analyte concentration increases over time in a linear gradient. We report here the application of nonlinear analyte concentration gradients for determining kinetic rates and equilibrium binding affinities in a single experiment. A versatile nonlinear gradient maker is presented, which is easily applied to microfluidic systems. Simulations validate that accurate kinetic rates can be extracted for a wide range of association and dissociation rates, gradient slopes, and curvatures, and with models for mass transport. The nonlinear analyte gradient method is demonstrated with a silicon photonic microring resonator platform to measure prostate specific antigen-antibody binding kinetics.
Department of Chemistry, University of Illinois Urbana-Champaign, Urbana, Illinois 61801, United States.
Michael T
MT
Marty
Courtney D Kuhnline
CD
Sloan
Ryan C
RC
Bailey
Stephen G
SG
Sligar
D055910:Antibodies, Immobilized / Q000276:immunology*; D000937:Antigen-Antibody Reactions; D015374:Biosensing Techniques / Q000295:instrumentation* / Q000379:methods; D003198:Computer Simulation; D004867:Equipment Design; D006801:Humans; D007700:Kinetics; D008954:Models, Biological; D017711:Nonlinear Dynamics; D017430:Prostate-Specific Antigen / Q000276:immunology*; D017186:Titrimetry / Q000295:instrumentation / Q000379:methods
D055910:Antibodies, Immobilized; D017430:Prostate-Specific Antigen