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Construction of Enzymosomes: Optimization of Coupling Parameters

C.A. Vale, M.L. Corvo, L.C.D. Martins, C.R. Marques, G. Storm, M.E.M. Cruz and M.B.F. Martins
Laboratorio Nacional de Engenharia, Tecnologia e Inovacao, IP, PT

enzymosomes, bioconjugates, lipid vesicles, therapeutic proteins, protein delivery

Different strategies can be used to improve carrier mediated delivery of therapeutic proteins. The incorporation of therapeutic enzymes into polymeric carriers [1], into aqueous space of lipid vesicles [2] or lipid-detergent vesicles [3, 4] or incorporation of hydrophobized forms into lipid bilayer of vesicles [5, 6, 7] are examples.
Another strategy, not usually used for therapeutic enzymes, is its attachment to the outer surface of liposomes, using technologies developed for antibodies [8, 9]. Few publications report the construction of carriers with surface attached enzymes [10] in contrast with a huge number of publications with antibodies attached to carriers, a concept widely used for the active targeting of nanocarriers.
In this work, we optimized processes to link covalently enzymes to the surface of lipid carriers, based on techniques of attachment of antibodies to liposomes. Our goal and concept is to build enzymosomes, as a tailor made load of therapeutic enzymes to liposome surface, to modulate the enzyme therapeutic effect by optimized carrier mediated transport. We achieve preservation of enzyme activity and preservation of vesicles structural integrity, as shown by in vivo fate.
The increasing number of potentially active proteins and its particular structural complexity are reasons for significance of this work to the drug delivery field.

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