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


RESULTS
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.


CONCLUSIONS
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.
Colletier
Jacques-Philippe
JP
Laboratoire de Synthèse et Physicochimie des Molécules d'Intérêt Biologiques-Groupe de Biochimie des Protéines, Université Paul Sabatier, Toulouse, France. Colletie@ibs.fr
Chaize
Barnabé
B
Winterhalter
Mathias
M
Fournier
Didier
D
D000110:Acetylcholinesterase / Q000737:chemistry / Q000378:metabolism; D000818:Animals; D001709:Biotechnology / Q000379:methods*; D002021:Buffers; D003080:Cold Temperature; D004798:Enzymes / Q000737:chemistry / Q000378:metabolism*; D005079:Excipients / Q000494:pharmacology; D008051:Lipid Bilayers / Q000737:chemistry / Q000378:metabolism*; D008081:Liposomes / Q000737:chemistry / Q000378:metabolism*; D010743:Phospholipids / Q000737:chemistry / Q000378:metabolism*
D002021:Buffers; D004798:Enzymes; D005079:Excipients; D008051:Lipid Bilayers; D008081:Liposomes; D010743:Phospholipids; D000110:Acetylcholinesterase