Abstract
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 μm 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 I–V characteristics, fast response times, good linear sensitivity (0.103 nA mm−2 mM−1, R2 = 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.
Highlights
High resolution micropatterning of functional proteins is an essential tool in the design of many new diagnostic and therapeutic strategies
The results indicate that the biotin binding capability of self-aligned strep tavidin is preserved, as well as the catalytic activity of the biotinylatedHRP introduced by the biotin-avidin binding precisely onto the micro electrode array
We demonstrate a rapid and high-resolution geometric protein self-patterning (GPS) method for effective immobilization of protein molecules onto a microelectrode array supported by a hydrophobic perfluoropolymer surface for the fabrication of microelectronic biosensors
Summary
High resolution micropatterning of functional proteins is an essential tool in the design of many new diagnostic and therapeutic strategies. A microelectrode with the dimension of tens of micrometers possesses a hemispherical or cylindrical diffusion flux of analyte with enhanced mass transport rates, which results in improved electrochemical kinetics and analytical performance, such as faster response, higher sensitivity and wider dy namic range, compared with a macroelectrode where planar diffusion predominates (Matsui et al, 2017; Pemberton et al, 2011; Aoki et al, 1987; Kovach et al, 1985; Streeter et al, 2007; Ito et al, 1972; Compton et al, 2008) These superior characteristics are important in wearable and implantable biosensors for real-time monitoring. An array of multiple microelectrodes (microelectrode array) is often used to overcome the low electric current associated with a single microelectrode (Li et al, 2019; Chen and White (2011); Hintsche et al, 1994)
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