Nonlinear analyte concentration gradients for one-step kinetic analysis employing optical microring resonators

Michael T. Marty, Courtney D.Kuhnline Sloan, Ryan C. Bailey, Stephen G. Sligar

Research output: Contribution to journalArticlepeer-review

14 Scopus citations

Abstract

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.

Original languageEnglish (US)
Pages (from-to)5556-5564
Number of pages9
JournalAnalytical Chemistry
Volume84
Issue number13
DOIs
StatePublished - Jul 3 2012
Externally publishedYes

ASJC Scopus subject areas

  • Analytical Chemistry

Fingerprint

Dive into the research topics of 'Nonlinear analyte concentration gradients for one-step kinetic analysis employing optical microring resonators'. Together they form a unique fingerprint.

Cite this