Reflecting recent work in the The Futaki Lab

Published in Bioconjugate Chemistry, researchers from Shiroh Futaki Group at the Kyoto University, prepared conjugates of the biocompatible polysaccharide pullulan with a cell membrane permeabilizing peptide L17E, PL-L17Es, with the aim of producing complex coacervates with pronounced intracellular antibody, IgG, delivery activity and stable structures. Coacervates with diameters of a few μm were formed simply by mixing PL-L17Es with IgG labeled with negatively charged fluorescent moieties of Alexa Fluor 488 [IgG(AF488)].

The coacervate resulted in a pronounced cytosolic infusion of IgG(AF488) and IgG binding to the target proteins inside the cell. The droplet structures were maintained even under high salt conditions, and the fluorescence in the droplet was not recovered after photobleaching, suggesting the formation of complex coacervate microgels.

Dynamic changes in cell membrane structure to entrap the coacervate microgels were captured by confocal and electron microscopy, resulting in cytosolic IgG infusion. The use of M-lycotoxin instead of L17E resulted in a coacervate microgel with marked IgG delivery activity even in the presence of serum.

Successful IgG delivery to primary hepatocytes, undifferentiated induced pluripotent stem, iPS, cells, and iPS cell-derived intestinal epithelial cells was also achieved. The construction of complex coacervate microgels with design flexibility and the validity of intracellular IgG delivery with high salt stability were thus demonstrated.