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- Title
Fully automatic flow-based device for monitoring of drug permeation across a cell monolayer.
- Authors
Zelená, Lucie; Marques, Sara; Segundo, Marcela; Miró, Manuel; Pávek, Petr; Sklenářová, Hana; Solich, Petr
- Abstract
A novel flow-programming setup based on the sequential injection principle is herein proposed for on-line monitoring of temporal events in cell permeation studies. The permeation unit consists of a Franz cell with its basolateral compartment mixed under mechanical agitation and thermostated at 37 °C. The apical compartment is replaced by commercially available Transwell inserts with a precultivated cell monolayer. The transport of drug substances across epithelial cells genetically modified with the P-glycoprotein membrane transporter (MDCKII-MDR1) is monitored on-line using rhodamine 123 as a fluorescent marker. The permeation kinetics of the marker is obtained in a fully automated mode by sampling minute volumes of solution from the basolateral compartment in short intervals (10 min) up to 4 h. The effect of a P-glycoprotein transporter inhibitor, verapamil as a model drug, on the efficiency of the marker transport across the cell monolayer is thoroughly investigated. The analytical features of the proposed flow method for cell permeation studies in real time are critically compared against conventional batch-wise procedures and microfluidic devices.
- Subjects
FLUID flow; SEQUENTIAL injection analysis; DRUG design; GLYCOPROTEINS; RHODAMINES
- Publication
Analytical & Bioanalytical Chemistry, 2016, Vol 408, Issue 3, p971
- ISSN
1618-2642
- Publication type
Article
- DOI
10.1007/s00216-015-9194-0