Preparative size exclusion chromatography combined with detergent removal as a versatile tool to prepare unilamellar and spherical liposomes of highly uniform size distribution |
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| Authors: | Martin Holzer Sabine BarnertJoachim Momm Rolf Schubert |
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| Affiliation: | Department of Pharmaceutical Technology and Biopharmacy, University of Freiburg, Hermann-Herder-Strasse 9, D-79104 Freiburg, Germany |
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| Abstract: | Detergent removal from mixed micelles was combined with preparative size exclusion chromatography (SEC) on Sephacryl S 500 HR to prepare unilamellar and spherical liposomes of defined sizes between 50 and 100 nm with a very narrow size distribution (RSD of vesicle diameter between 13% and 25%). For neutral phosphatidylcholine and negatively charged phosphatidylcholine/phosphatidylglycerol liposome preparations, efficient sizing at the preparative scale was demonstrated by analyzing isolated SEC peak fractions with cryo-transmission electron microscopy and dynamic light scattering. The number-weighted average vesicle diameters obtained using both methods are in very good agreement for fractions of low polydispersity. |
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| Keywords: | SEC, size exclusion chromatography DLS, dynamic light scattering cryo-TEM, cryo-transmission electron microscopy EPC, egg-phosphatidylcholine OG, octyl-β-d-glucopyranoside SC, sodium cholate DPH, diphenylhexatriene RFU, relative fluorescence units EPC-90, 100% EPC liposomes, average DLS diameter 90 nm EPC-60, 100% EPC liposomes, average DLS diameter 60 nm EPCPG-90, EPC:EPG 90:10 mol% liposomes, average DLS diameter 90 nm EPCPG-60, EPC:EPG 90:10 mol% liposomes, average DLS diameter 60 nm |
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