(293f) Antigen Specific Lysis: High Throughput, High Purity Cell Sorting Through Polymer Design

Lilly, J., University of Kentucky
Xu, W., University of South Carolina
Romero-Uribe, G., University of Kentucky
Balasubramaniam, V., Univeristy of Colorado at Denver School of Medicine
Berron, B. J., University of Colorado

High purity cell sorting is at the core of many basic and applied clinical research studies. Current approaches including fluorescence-activated cell sorting (FACS) and magnetic-activated cell sorting (MACS) are limited by low throughput and low specificity, respectively. A faster, high purity method of cell isolation would improve the quality and rate of downstream biological investigations.

Here, we describe a novel approach for high throughput cell isolation based the specific protection of targeted cells and the lysis of nonspecific species.  Briefly, antibody affinity directs the localization of a photoinitiator on the surface of targeted cells, the cells are immersed in monomer solution, and visible light induces cross-linking of poly(ethylene glycol) diacrylate (Mn=575) exclusively around target cells. Upon rinsing, the antigen-positive cells are now encapsulated by a crosslinked polymer film, where antigen-negative cells are unprotected.  The entire population is then exposed to lysis agents  (surfactants, proteases, hypotonic solutions, etc.) to lyse unprotected cells. Our initial results show encapsulated cells are >75% viable, where unprotected cells are >99.5% dead. We describe in detail the structure and behavior of the protective polymer and relate these characteristics to the exclusion of macromolecules and lysing agents.