Layer-by-layer self-assembled polyelectrolyte films containing a charged cyclodextrin and lipopolysaccharide (LPS) are developed for the first time as a potential model for local endotoxin antagonist delivery. We have examined the biological activity of a lipopolysaccharide from E. coli incorporated into multilayered architectures made of poly-(L-lysine) and poly-(L-glutamic acid). Used in such build-ups, a Polycationic cyclodextrin heptakis(6-deoxy-6-pyridylamino)-beta-cyclodextin showed molecular chaperone properties by enabling restoration of the LPS biological activity whenever lost upon interaction with poly-(L-lysine).