Cell-Penetrating Peptides as Carriers for Transepithelial Drug Delivery In Vitro
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 wordsCaco-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.
- 11.Boegh M, Baldursdottir SG, Nielsen MH, Müllertz A, Nielsen HM (2013) Development and rheological profiling of biosimilar mucus. Annu Trans Nord Rheol Soc 21:233–240Google Scholar
- 20.Nielsen EJB, Yoshida S, Kamei N, Iwamae R, Khafagy e-S, Olsen J, Rahbek UL, Pedersen BL, Takayama K, Takeda-Morishita M (2014) In vivo proof of concept of oral insulin delivery based on a co-administration strategy with the cell-penetrating peptide penetratin. J Control Release 189:19–24CrossRefPubMedGoogle Scholar
- 28.Tréhin R, Nielsen HM, Jahnke H-G, Krauss U, Beck-Sickinger AG, Merkle HP (2004) Metabolic cleavage of cell-penetrating peptides in contact with epithelial models: human calcitonin (hCT)-derived peptides, Tat(47-57) and penetratin(43-58). Biochem J 382:945–956PubMedCentralCrossRefPubMedGoogle Scholar