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

Abstract

This is an accepted version of a paper published in Analytical and Bioanalytical Chemistry. This paper has been peer-reviewed but does not include the final publisher proof-corrections or journal pagination.Citation for the published paper: Meiby, E., Morin Zetterberg, M., Ohlson, S., Agmo Hernańdez, V., Edwards, K. (2013 AbstractLipodisks, 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 containing15 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 for the potential to study ligand interactions with membrane proteins by weak affinity chromatography (WAC).

Label-Free Characterization of Peptide–Lipid Interactions Using Immobilized Lipodisks

“…The results reported in this study illustrate the analytical potential of immobilized lipodisks. In addition to previously reported interaction parameters of proteins and lipodisks, , the characterization of the affinity between lipodisks and low molecular weight peptides opens the possibility of studying the membrane interaction of smaller molecules. It also represents a big step toward the characterization of the binding of antimicrobial peptides of therapeutical relevance to potential delivery vehicles such as the lipodisks.…”

“…24 Following adequate protocols, lipodisks can be immobilized on a variety of substrates, including the quartz crystal microbalance (QCM) 5 and surface plasmon resonance (SPR) 25 sensors, as well as on chromatographic media. 5,26,27 This allows the study of, for example, protein−membrane interactions 5,25 and drug partition. 26 In this report, a reproducible and robust protocol for the covalent binding of lipodisks to sensors for QCM with dissipation monitoring (QCM-D) experiments is presented.…”

“…Recent research has also shown that they are potentially useful as drug carriers . Following adequate protocols, lipodisks can be immobilized on a variety of substrates, including the quartz crystal microbalance (QCM) and surface plasmon resonance (SPR) sensors, as well as on chromatographic media. ,, This allows the study of, for example, protein–membrane interactions , and drug partition …”

EGF-targeting lipodisks for specific delivery of poorly water-soluble anticancer agents to tumour cells

“…Since their discovery almost two decades ago, 14 the lipodisks have proven their utility as adaptable and versatile biomimetic membranes. [15][16][17] During the last couple of years these non-toxic, biocompatible structures have also received increasing attention as novel drug carriers.…”

Lipodisks integrated with weak affinity chromatography enable fragment screening of integral membrane proteins

“…22 The usefulness of immobilized lipodisks is illustrated by recent reports concerning the study of lipid-peptide interactions, 19,23 drugmembrane partitioning 22,[24][25][26] and the binding of the peripheral membrane protein COX-1. 18,27 In the present report we take the lipodisk-concept one step further and demonstrate for the first time how the interactions between an integral membrane protein and small molecules can be successfully studied using proteolipodisks covalently coupled to chromatographic media.…”