24 August 2010 Parallel microfluidic arrays for SPRi detection
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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. 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 alpha-thrombin. The device consists of 264 element-addressable chambers of 700 pL each isolated by microvalves. The device also contains a dilution network for simultaneous interrogation of up to six different target concentrations, further speeding detection times. The element-addressable design of the array allows interrogation of multiple ligands against multiple targets, and analytes from individual chambers may be collected for downstream analysis. We demonstrate methods for educing nonspecific binding to the sensor surface and quantify the success of these methods using mass spectrometric identification of proteins eluted from our microfluidic chambers following SPRi analysis of crude cell lysates.
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Eric T. Lagally, Eric T. Lagally, Eric Ouellet, Eric Ouellet, Chris Lausted, Chris Lausted, Tao Lin, Tao Lin, Cheng-Wei Tony Yang, Cheng-Wei Tony Yang, Helen L. Lund, Helen L. Lund, Leroy Hood, Leroy Hood, } "Parallel microfluidic arrays for SPRi detection", Proc. SPIE 7759, Biosensing III, 77590J (24 August 2010); doi: 10.1117/12.861434; https://doi.org/10.1117/12.861434
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