Precise and rapid solvent-assisted geometric protein self-patterning with submicron spatial resolution for scalable fabrication of microelectronic biosensors

We report a novel manufacturing method for microelectronic biosensors with a one-step wet/de-wet micropatterning of proteins onto printed metal microelectrode array supported by a hydrophobic perfluoropolymer substrate. The method exploits the phenomenon that protein adsorption toward a hydrophobic perfluoropolymer surface is drastically suppressed by self-assembly of proteins into micelles in a solvent-buffer mixture, followed by a rapid wet/de-wet process to localize the protein molecules onto the hydrophilic microelectrodes. The method affords protein micropatterning with 5 mm spatial resolution while preserving biological function and is applicable to various proteins such as albumin, enzymes, antibodies and avidin . By combining the protein micropatterning with metal nanoparticle chemisorption printing of the microelectrode array, we demonstrate as a model, facile fabrication of a microelectronic glucose biosensor supporting convergent analyte diffusion and thus showing typical steady-state I-V characteristics and fast steady-state current response (~20-60 s) together with an ultra-wide linear dynamic range (2-100 mM). Our findings provide a new technical solution for precise and accurate coupling of biomolecules with a microelectronic array interface with important implications for the scale up and manufacture of diagnostics, biofuel cells and bioelectronic devices.