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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.

Original publication

DOI

10.1007/978-1-60761-919-2_9

Type

Journal article

Journal

Methods Mol Biol

Publication Date

2011

Volume

683

Pages

117 - 128

Keywords

Cell-Penetrating Peptides, Fluorescent Dyes, HeLa Cells, Humans, Intracellular Space, Kinetics, Protein Transport, Spectrometry, Fluorescence, Temperature