Cell-Penetrating Peptides as Carriers for Transepithelial Drug Delivery In Vitro

  • Stine Rønholt
  • Mie Kristensen
  • Hanne Mørck Nielsen
Part of the Methods in Molecular Biology book series (MIMB, volume 1324)


There is a growing interest in the use of cell-penetrating peptides (CPPs) as carriers for transepithelial drug delivery. This chapter gives an introduction to and discussion of the commonly used production and characterization methods for CPP–cargo samples including high-throughput cell viability screening. Moreover, we describe methods for permeation and cell viability assessment in the Caco-2 cell culture model with and without implementation of biosimilar mucus. Last, a method to assess metabolic degradation in vitro is described.

Key words

Caco-2 Cell viability Cell-penetrating peptides as carriers High-throughput screening In vitro Mucus Metabolic degradation Permeability Transepithelial drug delivery 



María García-Díaz is acknowledged for supplying the protocols for the mucus preparation and the permeation study using Caco-2 cell monolayers supplemented with biosimilar mucus. The research leading to these results has received support from the Innovative Medicines Initiative Joint Undertaking under grant agreement no. 115363 resources which are composed of financial contribution from the European Union’s Seventh Framework Programme (FP7/2007–2013) and EFPIA companies’ in kind contribution. Additionally, the work was financially supported by the Drug Research Academy (University of Copenhagen), The Danish Agency for Science, Technology & Innovation, and the Novo Nordisk Foundation Center for Protein Research.


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Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  • Stine Rønholt
    • 1
  • Mie Kristensen
    • 1
  • Hanne Mørck Nielsen
    • 2
  1. 1.Department of Pharmacy, Faculty of Health and Medical SciencesUniversity of CopenhagenCopenhagenDenmark
  2. 2.Section for Biologics, Department of Pharmacy, Faculty of Health and Medical SciencesUniversity of CopenhagenCopenhagenDenmark

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