Thermosensitive display of carbohydrate ligands on microgels for switchable binding of proteins and bacteria.

Abstract

The synthesis of carbohydrate-functionalized thermosensitive poly(N-isoproylacrylamide) microgels and their ability to bind carbohydrate-binding pathogens upon temperature switch is reported. It is found that the microgels binding affinity is increased above their lower critical solution temperature (LCST) enabling thermo-triggerable capture of pathogens. Here a series of microgels with comparatively low mannose functionalization degrees below 1 mol % is achieved by a single polymerization step. Upon increase of mannose density, the microgel size increases and the LCST decreases to 26°C. Clustering with concanavalin A indicated that binding affinity is enhanced by a higher mannose content and by raising the temperature above the LCST. Binding studies with E. coli confirm stronger specific interactions above the LCST and formation of mechanically stable aggregates enabling efficient separation of E. coli by filtration. For small incubation times above LCST, the microgels potential to release pathogens again below LCST is confirmed as well. Compared to existing switchable scaffolds, microgels nearly entirely composed of thermosensitive material undergo a large change in volume, which allows them to drastically vary the density of ligands to switch between capture and release. This is straightforward, yet novel approach is likely compatible with a broad range of bioactive ligands. Therefore, thermosensitive microgels represent a promising platform for the specific capture or release of cells or pathogens.