Immobilized lipodisks as model membranes in high-throughput HPLC-MS analysis
2013 (English)In: Analytical and Bioanalytical Chemistry, ISSN 1618-2642, E-ISSN 1618-2650, Vol. 405, no 14, 4859-4869 p.Article in journal (Refereed) Published
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.
Place, publisher, year, edition, pages
2013. Vol. 405, no 14, 4859-4869 p.
Lipodisks, COX-1, HPLC-MS, Model membrane, Drug partition studies, Membrane protein, WAC, Weak affinity chromatography
IdentifiersURN: urn:nbn:se:uu:diva-197337DOI: 10.1007/s00216-013-6892-3ISI: 000318312400018OAI: oai:DiVA.org:uu-197337DiVA: diva2:612605
FunderSwedish Research Council
De två (2) första författarna delar förstaförfattarskapet.2013-03-222013-03-222016-03-09Bibliographically approved