Label-free focusing of viral particles under a temperature gradient coupled with continuous swirling flow

The advances of biomedicine and biotechnology demand new approaches to enrich biological nanoparticles, such as viruses, viral vectors and nanovesicles, in an easy-to-operate fashion. Conventional methods, such as ultracentrifugation and ultrafiltration, require bulky instruments and extensive manual operation. Inspired by recent research of thermophoresis of biomolecules and bio-nanoparticles in aqueous solutions, we present a microfluidic design that directly focuses nanoparticles in a label-free and flow-through process by coupling an engineered swirling flow and a moderate, one-dimensional temperature gradient. Enrichment of polystyrene particles, HIV and bacteriophage samples was quantitatively determined, indicating the compatibility of the microfluidic approach with synthetic and biological samples. The focusing results are well predicted using a numerical model. As thermophoresis is ubiquitous, the microfluidic approach can be applied broadly to bio-nanoparticle enrichment without the necessity of labeling, buffer exchange, or sheath fluids, permitting continuous retrieval of concentrated species in a simple, controlled flow with little infrastructure needs.