Enhanced Green Fluorescent Protein Streaming Dielectrophoresis in Insulator-Based Microfluidic Devices

There is tantalizing evidence that proteins can be accurately and selectively manipulated by higher order electric field effects within microfluidic devices. The accurate and precise manipulation of proteins in these platforms promises to disrupt and revolutionize many fields, most notably analytical biochemistry. Several lines of experimental evidence suggest much higher forces are generated compared to those calculated from traditional theories and those higher forces arise from subtle structural features of the proteins providing selectivity. New theories reflect some of the experimental evidence in the magnitude of the force predicted and inclusion of subtle structural features absent in traditional continuum theory. Unfortunately, the experimental evidence is largely exploratory in nature and lacks one or more important elements that prevents a clear interpretation and comparison to not only the other existing data, but also quantitative comparison to the evolving theoretical descriptions. Here, a clear and interpretable experimental system is presented that quantitatively determines the dielectrophoretic susceptibility of unlabeled, unaggregated native-structure protein molecules that are exposed to modest electric fields (10⁵-10⁶ V/m) for short periods of time (∼5 ms) without significant increases in local concentration. The platform uses sub-nanogram quantities of protein, the probed volume upon determination is a few picoliters, and the total analysis time is 10 s.