Herein we will describe a new surface modification approach for isolating high purity cell populations based on the use of a polymeric coating that stabilize cells against a variety of destructive mechanisms (detergent, enzymes, shear, hypotonic stress), enabling a variety of novel selective lysis and degradation techniques for specific cell isolation. We tailored PEG diacrylate polymer coatings which are formed in response to surface marker expression and are capable of stabilizing Jurkat cells in response to immersion in deionized water. Specific antibodies against CD45 were used to specifically label the surface of Jurkat cells. Then the cells immersed in photoinitiators conjugated to streptavidin, immobilizing polymerization initiators on the surface of these specifically labeled cells. Upon exposure to PEG diacrylate and 530 nm light, a crosslinked polymer coating is formed on the outside of the initiator labeled cells, forming a synthetic extracellular membrane in response to expression of CD45. After rinsing in phosphate buffer, coated cells were resuspended in pure deionized water and were intact and of spherical morphology. Cell viability assays confirm the coated cells are largely viable (> 80%) after polymerization and after immersion in water (~75%). In the absence of a polymer coating, < 0.5% of the cells are viable after 10 minutes in water. The drastic contrast in stability of coated cells from their native counterparts supports Antigen Specific Lysis as a novel paradigm for cell isolation based on sensitive, specific coating formation.

Antigen specific lysis for large-volume, high-purity cell isolation

246th National Meeting (2013)

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