title: Construction of a liposome dialyzer for the preparation of high-value, small-volume liposome formulations.
abstract: The liposome dialyzer is a small-volume equilibrium dialysis device, built from commercially available materials, that is designed for the rapid exchange of small volumes of an extraliposomal reagent pool against a liposome preparation. The dialyzer is prepared by modification of commercially available dialysis cartridges (Slide-A-Lyzer cassettes), and it consists of a reactor with two 300-μl chambers and a 1.56-cm(2) dialysis surface area. The dialyzer is prepared in three stages: (i) disassembling the dialysis cartridges to obtain the required parts, (ii) assembling the dialyzer and (iii) sealing the dialyzer with epoxy. Preparation of the dialyzer takes ∼1.5 h, not including overnight epoxy curing. Each round of dialysis takes 1-24 h, depending on the analyte and membrane used. We previously used the dialyzer for small-volume non-enzymatic RNA synthesis reactions inside fatty acid vesicles. In this protocol, we demonstrate other applications, including removal of unencapsulated calcein from vesicles, remote loading and vesicle microscopy.
authors.affiliation: MIT Media Lab, Cambridge, Massachusetts, USA.
authors.forename: Katarzyna
authors.initials: K
authors.lastname: Adamala
authors.affiliation: Department of Molecular Biology, and Center for Computational and Integrative Biology, Howard Hughes Medical Institute, Massachusetts General Hospital, Boston, Massachusetts, USA.
authors.forename: Aaron E
authors.initials: AE
authors.lastname: Engelhart
authors.affiliation: Department of Molecular Biology, and Center for Computational and Integrative Biology, Howard Hughes Medical Institute, Massachusetts General Hospital, Boston, Massachusetts, USA.
authors.forename: Neha P
authors.initials: NP
authors.lastname: Kamat
authors.affiliation: Department of Molecular Biology, and Center for Computational and Integrative Biology, Howard Hughes Medical Institute, Massachusetts General Hospital, Boston, Massachusetts, USA.
authors.forename: Lin
authors.initials: L
authors.lastname: Jin
authors.affiliation: Department of Molecular Biology, and Center for Computational and Integrative Biology, Howard Hughes Medical Institute, Massachusetts General Hospital, Boston, Massachusetts, USA.
authors.forename: Jack W
authors.initials: JW
authors.lastname: Szostak
mesh_terms: D002626:Chemistry, Pharmaceutical; D003956:Dialysis / Q000295:instrumentation; D008081:Liposomes / Q000737:chemistry*
chemical_list: D008081:Liposomes