The Phospholipid Vesicle-Based Drug Permeability Assay: 5. Development Toward an Automated Procedure for High-Throughput Permeability Screening
ForfatterFlaten, Gøril Eide; Awoyemi, Opeyemi Linda Ronke; Luthman, Kristina; Brandl, Martin; Massing, Ulrich
In-vitro screening for oral absorption has become an essential part of drug discovery and development. Recently, a new phospholipid vesicle-based permeation assay was developed which has shown to satisfyingly predict passive absorption of drugs in humans. The purpose of the current study was to investigate whether the assay may be further developed into a high-throughput tool by automating its most time consuming steps. The following challenges were addressed: (i) to design, build and test a heatsealing machine for mounting of the desired type of filter support onto both single wells and 24-well titre plate inserts, and (ii) to transfer the permeability assay to a robotic work station with attached UV-reader. The workstation is able to pipette and transport both plates and filter inserts and perform on-line photometric quantification of the amount of drug permeated. In order to enable the robot to move single (Standard-Transwell®) filter inserts, an extension of the gripping arm was designed, built and tested. Furthermore, in an alternative approach 24-well filter plates (Millicell®) were used instead of single filter inserts. The latter turned out to be more suitable in terms of error-free high-throughput robotic handling. The permeability values of drugs gained by the two automated procedures were compared with those measured by manual handling of the assay. Only neglectable differences in permeability values were seen. In conclusion, the most time-consuming steps of the assay were shown to be eligible for automation. This represents an interesting addition to the tool-box of in-vitro permeability screening assays running in a medium- to high-throughput format due to its easiness, its transferability to other laboratories and its good correlation with in vivo data on fraction absorbed of drugs in humans.
This is the accepted manuscript version. Published version available at http://dx.doi.org/10.1016/j.jala.2008.04.002
SiteringJournal of the Association for Laboratory Automation 14(2009) nr. 1 s. 12-21
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