Reaching for the limits in continuous-flow dielectrophoretic DNA analysis.

The efficient purification and analysis of topological DNA variants is mandatory for many state-of-the-art molecular medicine technologies, like gene- and cancer-therapy as well as plasmid vaccination. In this work, we exploit dielectrophoresis (DEP) for a fast and efficient continuous-flow separation and analysis that goes beyond the standard methods of gel electrophoresis and capillary electrophoresis. The aim of this work was to reach for the limits in dielectrophoretic analysis of DNA regarding the size resolution and the topological conformation. A continuous-flow analytical separation of analyte mixtures of small linear DNA-fragments (10.0 kbp, 8.0 kbp, 6.0 kbp, and 5.0 kbp) and topological DNA variants (linear and supercoiled conformation) was investigated. We present a world record in the minimal size difference of 16.7% of DNA samples that can be resolved in a dielectrophoretic continuous-flow separation. Moreover, we demonstrate for the first time a microfluidic continuous-flow separation of DNA molecules based on their topological conformation. Since dielectrophoresis is virtually label-free, it offers a fast in-process quality control with low consumption, e.g. for the production of gene vaccines.