Construction of a liposome dialyzer for the preparation of high-value, small-volume liposome formulations

Katarzyna Adamala; Aaron E. Engelhart; Neha P. Kamat; Lin Jin; Jack W. Szostak

doi:10.1038/nprot.2015.054

Construction of a liposome dialyzer for the preparation of high-value, small-volume liposome formulations

Katarzyna Adamala, Aaron E. Engelhart, Neha P. Kamat, Lin Jin, Jack W. Szostak^*

^*Corresponding author for this work

Biomedical Engineering

Research output: Contribution to journal › Article › peer-review

15 Scopus citations

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-cm² 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.

Original language	English (US)
Pages (from-to)	927-938
Number of pages	12
Journal	Nature Protocols
Volume	10
Issue number	6
DOIs	https://doi.org/10.1038/nprot.2015.054
State	Published - Jun 30 2015

Funding

acknoWleDGMents This work was supported in part by The National Aeronautics and Space Administration (NASA) Exobiology grant NNX07AJ09G to J.W.S. and in part by a grant (290363) from the Simons Foundation to J.W.S. A.E.E. and N.P.K. were supported by appointments to the NASA Postdoctoral Program, administered by Oak Ridge Associated Universities through a contract with NASA. A.E.E. was supported by a Tosteson Fellowship from the Massachusetts General Hospital Executive Committee on Research. J.W.S. is an Investigator of the Howard Hughes Medical Institute.

ASJC Scopus subject areas

General Biochemistry, Genetics and Molecular Biology

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10.1038/nprot.2015.054

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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-cm2 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.",

author = "Katarzyna Adamala and Engelhart, {Aaron E.} and Kamat, {Neha P.} and Lin Jin and Szostak, {Jack W.}",

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Construction of a liposome dialyzer for the preparation of high-value, small-volume liposome formulations

Abstract

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