Abstract
Lipid nanoparticles composed of mixtures of PEGylated-lipids; cationic and neutral lipids prepared by detergent dialysis can encapsulate biological active molecules and show considerable potential as systemic therapeutic agents. Addition of biotinylated lipids to this formulation allows surface modification of these particles with a suitable ligand or probe conjugated to streptavidin for specific cell targeting. Monitoring long circulating particles and cellular uptake requires stable and bright fluorescent probes. Quantum dots (QDs) constitute a relatively new class of fluorescent probes that overcome the limitations of organic fluorophores in biological imaging applications. Here, a protocol for the encapsulation of QD655 (red) in biotinylated lipid particles (BLPs) prepared by a detergent dialysis technique is presented followed by characterization of the loaded liposomal vehicles. Then, a protocol for BLPs surface modification via biotin-streptavidin linkage with preformed complexes of ligand-QD525 (green) for specific cell targeting of the nanoparticle is detailed. Conditions for cell binding and uptake of two colors QD labeled BLPs as well as basic microscopic settings for confocal live cell imaging are described.
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Acknowledgements
This work was financially supported by the Max Planck Society, the DFG Research Center for Molecular Physiology of the Brain (CMPB)—Excellence Cluster EXC 171—FZT103 “Microscopy at the Nanometer Range,” and the EU sixth framework (FP6) project FLUOROMAG, LHSB-CT-2006-037465.
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Sigot, V. (2014). Assembly, Characterization, and Delivery of Quantum Dot Labeled Biotinylated Lipid Particles. In: Fontes, A., Santos, B. (eds) Quantum Dots: Applications in Biology. Methods in Molecular Biology, vol 1199. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-1280-3_9
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DOI: https://doi.org/10.1007/978-1-4939-1280-3_9
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