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Open Access Highly Accessed Open Badges Methodology article

Protein encapsulation in liposomes: efficiency depends on interactions between protein and phospholipid bilayer.

Jacques-Philippe Colletier12, Barnabé Chaize12, Mathias Winterhalter2 and Didier Fournier1*

Author Affiliations

1 Laboratoire de Synthèse et Physicochimie des Molécules d'Intérêt Biologiques – Groupe de Biochimie des Protéines, Université Paul Sabatier, Toulouse, France

2 Institut de Pharmacologie et de Biologie Structurale – Laboratoire de Biophysique Membranaire, Toulouse, France

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BMC Biotechnology 2002, 2:9  doi:10.1186/1472-6750-2-9

Published: 10 May 2002



We investigated the encapsulation mechanism of enzymes into liposomes. The existing protocols to achieve high encapsulation efficiencies are basically optimized for chemically stable molecules. Enzymes, however, are fragile and encapsulation requires in addition the preservation of their functionality. Using acetylcholinesterase as a model, we found that most protocols lead to a rapid denaturation of the enzyme with loss in the functionality and therefore inappropriate for such an application. The most appropriate method is based on lipid film hydration but had a very low efficiency.


To improve it and to propose a standard procedure for enzyme encapsulation, we separate each step and we studied the effect of each parameter on encapsulation: lipid and buffer composition and effect of the different physical treatment as freeze-thaw cycle or liposomes extrusion. We found that by increasing the lipid concentration, increasing the number of freeze-thaw cycles and enhancing the interactions of the enzyme with the liposome lipid surface more than 40% of the initial total activity can be encapsulated.


We propose here an optimized procedure to encapsulate fragile enzymes into liposomes. Optimal encapsulation is achieved by induction of a specific interaction between the enzyme and the lipid surface.