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

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. / <p>De två (2) första författarna delar förstaförfattarskapet.</p>

Identiferoai:union.ndltd.org:UPSALLA1/oai:DiVA.org:uu-197337
Date January 2013
CreatorsMeiby, Elinor, Morin Zetterberg, Malin, Ohlson, Sten, Agmo Hernández, Víctor, Edwards, Katarina
PublisherUppsala universitet, Analytisk kemi, Uppsala universitet, Analytisk kemi, Uppsala universitet, Analytisk kemi, Department of Chemistry and Biomedical Sciences, Linnaeus University, Kalmar, Department of Chemistry and Biomedical Sciences, Linnaeus University, Kalmar Sweden AND School of Biological Sciences, Nanyang Technological University, Singapore
Source SetsDiVA Archive at Upsalla University
LanguageEnglish
Detected LanguageEnglish
TypeArticle in journal, info:eu-repo/semantics/article, text
Formatapplication/pdf
Rightsinfo:eu-repo/semantics/openAccess
RelationAnalytical and Bioanalytical Chemistry, 1618-2642, 2013, 405:14, s. 4859-4869

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