Biomolecular recognition on well-characterized beads packed in microfluidic channels

Analytical Chemistry
T BurandaG P Lopez

Abstract

We describe a new approach for the analysis of biomolecular recognition in microfluidic channels. The method involves real-time detection of soluble molecules binding to receptor-bearing microspheres, sequestered in affinity column format inside a microfluidic channel. Identification and quantitation of analytes occurs via direct fluorescence measurements or fluorescence resonance energy transfer (FRET). We establish a model system that detects the FLAG epitope. The assay can potentially detect subfemtomole quantities of antibody with a high signal-to-noise ratio and a large dynamic range spanning nearly 4 orders of magnitude in analyte concentration in microliter-to-submicroliter volumes of analyte fluid. Kinetic and equilibrium constants for the reaction of this receptor-ligand pair are obtained through modeling of kinetic responses of the affinity microcolumn and are consistent with those obtained by flow cytometry. Because of the correlation between kinetic and equilibrium data obtained for the microcolumns, quantitative analysis can be done prior to the steady-state end point of the recognition reaction. This method has the promise of combining the utility of affinity chromatography with the advantage of direct, quantitati...Continue Reading

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Citations

May 31, 2007·Chemical Society Reviews·Lourdes Basabe-DesmontsMercedes Crego-Calama
Jun 5, 2012·Biomicrofluidics·Cheng WangSascha Hilgenfeldt
Nov 24, 2007·Analytical and Bioanalytical Chemistry·Lourdes Basabe-DesmontsMercedes Crego-Calama
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