Protein patterning is ubiquitous in the creation of sensing motifs that rely on receptor-ligand binding for selectivity. Static microfluidic patterning (sMFP) has the potential to greatly aid in the development of simple, robust methods for deposition of biological capture agents onto a solid substrate. Traditionally, sMFP-based micro-immunoassays involving multiple binding regions suffer from the formation of an analyte depletion layer adjacent to the micro-channel floor where binding occurs. As a result, longer assay times are required to achieve equilibrium binding states for all binding surfaces. Here we present the use of grooved microchannels to induce mixing within the cross-section of sMFP microchannels, thus eliminating the occurrence of analyte depletion layers and consequently, decreasing the time required for equilibrium binding. The rate of mixing within the microchannel cross-section can be controlled via the geometries of the grooves themselves, which are easily modified through soft-lithography. The impact of different groove geometries, flow rates, and assay times on the immobilization density will be presented.

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