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    Immobilized lipodisks as model membranes in high-throughput HPLC-MS analysis

    Year: 2013

    Journal: Analytical and Bioanalytical Chemistry November 2012, 20130324

    Authors: Elinor Meiby 1, Malin Morin Zetterberg 2, Sten Ohlson 1 4, Víctor Agmo Hernández 2, Katarina Edwards 2 3

    Last authors: Katarina Edwards

    Organizations: 1. Department of Chemistry and Biomedical Sciences, Linnaeus University, 391 82, Kalmar, Sweden 2. Department of Chemistry—BMC, Uppsala University, Box 579, 751 23, Uppsala, Sweden 3. School of Biological Sciences, Nanyang Technological University, 60 Nanyang Drive, SBS-03n-28, Singapore, 637551, Singapore 4. FRIAS, School of Soft Matter Research, University of Freiburg, Albertstraße 19, 79104, Freiburg, Germany

    Country: sweden, Sverige, Singapore, Germany

    Lipodisks, also referred to as polyethylene glycol (PEG)-stabilized bilayer disks, have previously been demonstrated to hold great potential as model membranes in drug partition studies. In this study, an HPLC-MS system with stably immobilized lipodisks is presented. Functionalized lipodisks were immobilized on two different HPLC support materials either covalently by reductive amination or by streptavidin–biotin binding. An analytical HPLC column with immobilized lipodisks was evaluated by analysis of mixtures containing 15 different drug compounds. The efficiency, reproducibility, and stability of the system were found to be excellent. In situ incorporation of cyclooxygenase-1 (COX-1) in immobilized lipodisks on a column was also achieved. Specific binding of COX-1 to the immobilized lipodisks was validated by interaction studies with QCM-D. These results, taken together, open up the possibility of studying ligand interactions with membrane proteins by weak affinity chromatography.