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Uptake Kinetics of Cell-Penetrating Peptides

  • Anders Florén
  • Imre MägerEmail author
  • Ülo Langel
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 683)

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.

Key words

CPP uptake Kinetics Flow cytometry Fluorescence Bioluminescence Fluorescence quenching 

Notes

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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Copyright information

© Springer Science+Business Media, LLC 2011

Authors and Affiliations

  1. 1.Laboratory of Molecular Biotechnology, Institute of TechnologyTartu UniversityTartuEstonia
  2. 2.Department of NeurochemistryStockholm UniversityStockholmSweden

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