Liposome Immobilization on Cross-linked Polymer Gel by In Situ Formation of Cleavable Covalent Bonds

Abstract

Immobilization of liposomes onto chemically modified Sephacryl gel particles by in situreaction between liposome-incorporated thiols and mercapto moieties on the gel to form disulfide linkages was investigated. For the immobilization, both the mercapto moieties and the incorporated thiol were essential. The immobilization occurred upon coincubation of the modified liposomes with the modified gel for 48 hours. Once immobilized, no spontaneous detachment of the immobilized liposomes was observed. The degree of immobilization depended on both the thiol content and the ratio of the liposomes to the gel partilces. In a typical immobilization with 25mol% 1-octanethiol, 82% of the liposomal phosphatidylcholine in the system was found to be associated with the gel. By decreasing the ratio of the liposomes to gel it was possible to bring the immobilization close to quantitative one. Among the three different thiols examined (1-octanethiol, 1-hexadecanethiol and thiocholesterol), the extent of the immobilization was slightly higher with thiocholesterol than the alkanethiols. The immobilized liposomes were detached from the gel with dithiothreitol. Approximately 60% of the fluorescent dextran derivative encapsulated in the liposomes was retained throughout the immobilization-detachment process. The gel left after the detachment remained active for immobilizing a fresh batch of thiol-liposomes.