Mutiscale substrates based on hydrogel-incorporated silicon nanowires for protein patterning and microarray-based immunoassays

Here, protein micropatterns were prepared on micropatterned nanostructures for potential applications in microarray-based multiplex bioassays with enhanced protein-loading capacity and detection sensitivity. Vertically-aligned silicon nanowires (SiNWs) that were about 8 μm in height and 150 nm in diameter were prepared using an etching process and were surface-modified with aminopropyltriethoxysilane (APTES) to allow them to covalently immobilize proteins. The SiNW substrate was then overlaid with a micropattern of poly(ethylene glycol) (PEG) hydrogel to create defined arrays of microwells consisting of APTES-modified SiNW on the bottom of the wells, with hydrogel on the walls of the wells. Due to the non-adhesiveness of PEG hydrogels toward proteins, proteins were selectively immobilized on the surface-modified SiNW regions to create protein micropatterns. The increase in surface area increased the protein loading capacity of the SiNWs by more than 10 times the capacity of a planar silicon substrate. Immunobinding assays between IgG and anti-IgG and between IgM and anti-IgM that were performed on micropatterned SiNWs emitted stronger fluorescent signals and showed higher sensitivity than assays performed on planar silicon substrates. Finally, microfluidic channels were successfully integrated into the micropatterned SiNWs to enable the simultaneous performance of multiple immunoassays on a single microarray platform.

► Multiscale substrates were prepared by micropatterning hydrogel onto silicon nanowires.
► Protein patterning on the micropatterned nanostructures with higher protein-loading capacity.
► Highly-sensitive immunoassay was possible using resultant protein micropattern.
► Microfluidic device was integrated with micropatterned nanostructure for multiplex immunoassay.