In this work, we present a methodological approach to analyze an enhanced dielectrophoresis (DEP) system from both a circuit analysis and electrothermal view points. In our developed model, we have taken into account various phenomena and constraints such as voltage degradation (due to the presence of the protecting oxide layer), oxide breakdown, instrumentation limitations, and thermal effects. The results from this analysis are applicable generally to a wide variety of geometries and high voltage microsystems. Here, these design guidelines were applied to develop a robust electronic actuation system to perform a multiplexed bead-based protein assay. To carry out the multiplexed functionality, along a single microfluidic channel, an array of proteins is patterned, where each element is targeting a specific secondary protein coated on micron-sized beads in the subsequently introduced sample solution. Below each element of the array, we have a pair of addressable interdigitated electrodes. By selectively applying voltage at the terminals of each interdigitated electrode pair, the enhanced DEP, or equivalently 'ultra'-DEP (uDEP) force detaches protein-bound beads from each element of the array, one by one, without disturbing the bound beads in the neighboring regions. The detached beads can be quantified optically or electrically downstream. For proof of concept, we illustrated 16-plex actuation capability of our device to elute micron-sized beads that are bound to the surface through anti-IgG and IgG interaction which is on the same order of magnitude in strength as typical antibody-antigen interactions. In addition to its application in multiplexed protein analysis, our platform can be potentially utilized to statistically characterize the strength profile of biological bonds, since the multiplexed format allows for high throughput force spectroscopy using the array of uDEP devices, under the same buffer and assay preparation conditions.
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