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

Precise and high-resolution coupling of functional proteins with micro-transducers is critical for the manufacture of miniaturized bioelectronic devices. Moreover, electrochemistry on microelectrodes has had a major impact on electrochemical analysis and sensor technologies, since the small size of microelectrode affects the radial diffusion flux of the analyte to deliver enhanced mass transport and electrode kinetics. However, a large technology gap has existed between the process technology associated with such microelectronics and the conventional bio-conjugation techniques that are generally used. Here, we report on a high-resolution and rapid geometric protein self-patterning (GPS) method using solvent-assisted protein-micelle adsorption printing to couple biomolecules onto microelectrodes with a minimum feature size of 5 mu m and a printing time of about a minute. The GPS method is versatile for micropatterning various biomolecules including enzymes, antibodies and avidin-biotinylated proteins, delivering good geometric alignment and preserving biological functionality. We further demonstrated that enzyme-coupled microelectrodes for glucose detection exhibited good electrochemical performance which benefited from the GPS method to maximize effective signal transduction at the bio-interface. These microelectrode arrays maintained fast convergent analyte diffusion displaying typical steady-state characteristics, fast response times, good linear sensitivity (0.103 nA mm(-2) mM(-1), R-2 = 0.995) and an ultra-wide linear dynamic range (2-100 mM). Our findings provide a new technical solution for the precise and accurate coupling of biomolecules to a microelectronic array with important implications for the scaleup and manufacture of diagnostics, biofuel cells and bioelectronic devices that could not be realized economically by other existing techniques.

Automated summary

A new GPS method for coupling biomolecules onto microelectrodes has been developed, enabling high-resolution protein self-patterning. This versatile method can be used for micropatterning various biomolecules such as enzymes, antibodies, and avidin-biotinylated proteins.