High-Throughput Purification and Quantification of Bionanoparticles on Capillary-Channeled Fiber Stationary Phases

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With support from the Chemical Measurement and Imaging Program in the Division of Chemistry, Professor R. Kenneth Marcus and his group at Clemson University are working to prepare, characterize, and implement a unique format of fiber capillary-channeled polymers (C-CP) to facilitate the isolation, purification and quantification of biological and synthetic nanoparticles. Target systems include exosomes, a class of extracellular vesicles (EVs) that are integral components in intracellular communication, and thus are involved in normal homeostatic function, but also in the the evolution of abnormal biological function; thus there are long term implications of the studies being performed herein for biomedical science. The applications under study employ a hydrophobic interaction chromatography (HIC) protocol on polyester C-CP fiber columns, a system designed to probe aspects of fundamental biochemistry. The overall goal of the studies is to produce efficient, selective, and economical separations. This work is interdisciplinary, involving collaborative efforts among research groups in chemistry, bioengineering, mathematical, and materials sciences. If successful, the project may result in new commercial ventures in the textile and biopharmaceutical industries in South Carolina. Efforts related to developing a diverse workforce include integration of research topics into relevant coursework.The vast majority of separation/purification strategies for biological and synthetic nanoparticles involve physical means such as ultracentrifugation, ultrafiltration, and size-exclusion methods. Use of C-CP fibers to affect these separations rests on the ability to differentiate nanoparticles based upon chemical functionality, whose interaction strengths are determined in part based on particle size. The C-CP fiber platform can be implemented across a number of formats including spin-down micropipette tips and microbore columns. As a result, relevant problems in fundamental biochemistry, clinical analysis, and process analytical chemistry can be addressed. In-line particle sizing via multi-angle light scattering (MALS) provides both qualitative and quantitative information about eluted nanoparticles/vesicles. Surface modifications employing immobilized ligands can target selective capture of nanoparticles, complemented by selective immunolabelling strategies for species-specific assays. Multidimensional separations offer another innovative aspect for efficient characterization of nanoparticles from complex systems. In all cases, performance metrics will be compared to those available from commercial columns and standard methods. Development of practical methods will be augmented with fundamental studies directed at understanding physico-chemical processes at the microscopic level.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.