Abstract
As our knowledge increases about the diversity in uptake mechanisms displayed by cell-penetrating peptides (CPP), the concept of CPP uptake kinetics becomes increasingly complex. Here, we present three different assays that can be used for studying different kinetic aspects of CPP-mediated delivery: intracellular accumulation and membranolytical effects, intracellular CPP-cargo detachment, and finally a functional readout of a biological action from the delivered cargo. Unlike the traditional end-point measurements that give a static postincubation readout, these assays are all dynamic, real-time, in situ measurements obtained during incubation. A combination of some (or all) of these different assays gives us not only interesting kinetic information about the uptake routes but also provides a simple and valuable methodology for the evaluation of potential drug candidates based on the chemical modification of CPPs by cargo attachment.
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Acknowledgments
The work presented in this article was supported by: the Swedish Research Council (VR-NT); the Center for Biomem brane Research, Stockholm; the Knut and Alice Wallenberg’s Foundation; the EU through the European Regional Development Fund through the Center of Excellence in Chemical Biology, Estonia; the targeted financing SF0180027s08 from the Estonian Government; the DoRa Program of The European Social Fund; and the Archimedes Foundation.
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Florén, A., Mäger, I., Langel, Ü. (2011). Uptake Kinetics of Cell-Penetrating Peptides. In: Langel, Ü. (eds) Cell-Penetrating Peptides. Methods in Molecular Biology, vol 683. Humana Press. https://doi.org/10.1007/978-1-60761-919-2_9
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DOI: https://doi.org/10.1007/978-1-60761-919-2_9
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