Parallel microfluidic arrays for SPRi detection

Abstract

Surface Plasmon Resonance imaging (SPRi) is a label-free technique for the quantitation of binding affinities and concentrations for a wide variety of target molecules. Although SPRi is capable of determining binding constants for multiple ligands in parallel, current commercial instruments are limited to a single analyte stream and a limited number of ligand spots. Measurement of target concentration also requires the serial introduction of different target concentrations; such repeated experiments are conducted manually and are therefore time-intensive. Likewise, the equilibrium determination of concentration for known binding affinity requires long times due to diffusion-limited kinetics to a surface-immobilized ligand. We have developed an integrated microfluidic array using soft lithography techniques for SPRi-based detection and determination of binding affinities for DNA aptamers against human alphathrombin. The device consists of 264 element-addressable chambers isolated by microvalves. The resulting 700 pL volumes surrounding each ligand spot promise to decrease measurement time through reaction rate-limited kinetics. The device also contains a dilution network for simultaneous interrogation of up to six different target concentrations, further speeding detection times. Finally, the element-addressable design of the array allows interrogation of multiple ligands against multiple targets.