title: Immobilized lipodisks as model membranes in high-throughput HPLC-MS analysis.
abstract: 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.
authors.affiliation: Department of Chemistry and Biomedical Sciences, Linnaeus University, Kalmar, Sweden.
authors.forename: Elinor
authors.initials: E
authors.lastname: Meiby
authors.forename: Malin Morin
authors.initials: MM
authors.lastname: Zetterberg
authors.forename: Sten
authors.initials: S
authors.lastname: Ohlson
authors.forename: Víctor Agmo
authors.initials: VA
authors.lastname: Hernández
authors.forename: Katarina
authors.initials: K
authors.lastname: Edwards
mesh_terms: D040761:Biomimetic Materials / Q000737:chemistry*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D008051:Lipid Bilayers / Q000737:chemistry*; D008565:Membrane Proteins / Q000032:analysis / Q000737:chemistry*; D004364:Pharmaceutical Preparations / Q000032:analysis / Q000737:chemistry*; D011092:Polyethylene Glycols / Q000737:chemistry*; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*
chemical_list: D008051:Lipid Bilayers; D008565:Membrane Proteins; D004364:Pharmaceutical Preparations; D011092:Polyethylene Glycols