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Open Access Highly Accessed Methodology article

Liposome retention in size exclusion chromatography

Tristan Ruysschaert1, Audrey Marque1, Jean-Luc Duteyrat2, Sylviane Lesieur3, Mathias Winterhalter4 and Didier Fournier1*

Author Affiliations

1 Groupe de biotechnologie des protéines, Institut de Pharmacologie et de Biologie Structurale, F-31077, Toulouse, France

2 Electron Microscopy Department, Rangueil Hospital Medical School, University of Toulouse, 31062 Toulouse, France

3 Groupe Physico-chimie des systèmes polyphasés, Université Paris-sud, F-92296, Châtenay-Malabry, France

4 International University Bremen, D-28726 Bremen, Germany

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BMC Biotechnology 2005, 5:11  doi:10.1186/1472-6750-5-11

Published: 10 May 2005

Abstract

Background

Size exclusion chromatography is the method of choice for separating free from liposome-encapsulated molecules. However, if the column is not presaturated with lipids this type of chromatography causes a significant loss of lipid material. To date, the mechanism of lipid retention is poorly understood. It has been speculated that lipid binds to the column material or the entire liposome is entrapped inside the void.

Results

Here we show that intact liposomes and their contents are retained in the exclusion gel. Retention depends on the pore size, the smaller the pores, the higher the retention. Retained liposomes are not tightly fixed to the beads and are slowly released from the gels upon direct or inverted eluent flow, long washing steps or column repacking. Further addition of free liposomes leads to the elution of part of the gel-trapped liposomes, showing that the retention is transitory. Trapping reversibility should be related to a mechanism of partitioning of the liposomes between the stationary phase, water-swelled polymeric gel, and the mobile aqueous phase.

Conclusion

Retention of liposomes by size exclusion gels is a dynamic and reversible process, which should be accounted for to control lipid loss and sample contamination during chromatography.