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    Targeted detection of phosphatidylserine in biomimetic membranes and in vitro cell systems using annexin V-containing cubosomes

    Year: 2013

    Journal: Volume 34, Issue 33, November 2013, Pages 8361–8369, 20131003

    Authors: Hsin-Hui Shen 1 2, Vanessa Lake 3, Anton P. Le Brun 3, Michael James 4 5, Anthony P. Duff3 , Yong Peng 1, Keith M. McLean 1, Patrick G. Hartley 1

    Last authors: Patrick G. Hartley

    Organizations: 1 Materials Science and Engineering, CSIRO, Bayview Avenue, Clayton, VIC 3168, Australia 2 Department of Biochemistry and Molecular Biology, Monash University, Clayton, VIC 3168, Australia 3 The Bragg Institute, Australian Nuclear Science and Technology Organization (ANSTO), New Illawarra Rd, Lucas Heights, NSW 2234, Australia 4 The Australian Synchrotron, 800 Blackburn Rd, Clayton, VIC 3168, Australia 5 School of Chemistry, University of New South Wales, Kensington, NSW 2052, Australia

    Country: Australia

    In this work we have formulated Annexin V (ANX) decorated phosphatidylserine containing phytantriol (PSPhy) cubosomes to act as probes for the enhanced detection of apoptotic membranes in both model and in vitro cell systems. Small angle X-ray scattering (SAXS) and cryogenic-transmission electron microscopy (Cryo-TEM) indicated that ANX-containing PSPhy (ANX-PSPhy) cubosomes retain the Pn3m cubic symmetry and cubic phase nanoparticle characteristics of PSPhy cubosomes. The interaction of ANX-PSPhy cubosomes with apoptotic model and cellular membranes was also investigated using both quartz crystal microbalance with dissipation and confocal microscopy which confirmed that ANX-PSPhy cubosomes can selectively bind to apoptotic cells and model membranes. Neutron reflectometry has also been used to show strong binding of ANX-PSPhy cubosomes to a model apoptotic membrane, and in addition reveals changes in both the bilayer structure and in the internal structure of the cubosome in a region adjacent to the membrane as a result of material exchange. This material exchange between cubosome and apoptotic model bilayer was further demonstrated using Cryo-TEM. We have demonstrated that lipid bound protein, in this case Annexin V, can be used to target cubosome systems to biological surfaces in vitro.