Chapter 7
Quartz Crystal Microbalance with Dissipation Monitoring
(QCM-D): Preparing Functionalized Lipid Layers
for the Study of Complex Protein–Ligand Interactions
Holly L. Birchenough and Thomas A. Jowitt
Abstract
Quartz crystal microbalance with dissipation monitoring (QCM-D) is one of the most widely used
techniques for the deposition of lipid layers and provides a useful tool for protein–ligand analysis. By
using functionalized lipids, for example, with nitrilotriacetic acid (NTA) or biotin, one can couple a
molecule to the surface to investigate ligand interactions. Using lipid layers in this way allows for the
analysis of complex binding events such as conformational changes, fibrillation, and hierarchical clustering
on the surface, which is difficult to interpret with conventional surface sensor techniques. Deposition of
lipids and subsequent molecular interactions are easily monitored using both the frequency and the
dissipation, which have distinct features in bilayer formation and make QCM-D the ideal technique to
use. Here we describe the formation of biotinylated lipid bilayers using two different types of lipids and the
subsequent addition of avidin, which can then be used as a basis for linking biotinylated molecules to the
surface. These protocols can be adapted to use other lipid moieties and linking chemistries.
Key words Lipid bilayers, QCM-D, Functionalized lipids, Biosensors
1 Introduction
The utilization of lipid supports on sensor surfaces has increased
dramatically over the last few years. One reason for this is the
relative ease in which one can make a well-ordered layer that can
be tailored to suit the needs of the experiment. These can be
bilayers [1–5], self-assembled monolayers [6, 7], or tethered
bilayers, using the integration of other molecules as supports distancing the layer from the surface [8, 9]. The adaptability of the
approach has led to exciting new developments in
bio-nanotechnology [10, 11] and in the design of novel characterization methods for investigating molecular interactions, membrane
protein–lipid interactions, and antimicrobial peptides [3, 12–
14]. For the study of protein–ligand interactions, the layers can
be adapted with the addition of lipids with functional head groups
Tina Daviter et al. (eds.), Protein-Ligand Interactions: Methods and Applications, Methods in Molecular Biology, vol. 2263,
https://doi.org/10.1007/978-1-0716-1197-5_7, © Springer Science+Business Media, LLC, part of Springer Nature 2021
183
Quartz Crystal Microbalance with Dissipation Monitoring
(QCM-D): Preparing Functionalized Lipid Layers
for the Study of Complex Protein–Ligand Interactions
Holly L. Birchenough and Thomas A. Jowitt
Abstract
Quartz crystal microbalance with dissipation monitoring (QCM-D) is one of the most widely used
techniques for the deposition of lipid layers and provides a useful tool for protein–ligand analysis. By
using functionalized lipids, for example, with nitrilotriacetic acid (NTA) or biotin, one can couple a
molecule to the surface to investigate ligand interactions. Using lipid layers in this way allows for the
analysis of complex binding events such as conformational changes, fibrillation, and hierarchical clustering
on the surface, which is difficult to interpret with conventional surface sensor techniques. Deposition of
lipids and subsequent molecular interactions are easily monitored using both the frequency and the
dissipation, which have distinct features in bilayer formation and make QCM-D the ideal technique to
use. Here we describe the formation of biotinylated lipid bilayers using two different types of lipids and the
subsequent addition of avidin, which can then be used as a basis for linking biotinylated molecules to the
surface. These protocols can be adapted to use other lipid moieties and linking chemistries.
Key words Lipid bilayers, QCM-D, Functionalized lipids, Biosensors
1 Introduction
The utilization of lipid supports on sensor surfaces has increased
dramatically over the last few years. One reason for this is the
relative ease in which one can make a well-ordered layer that can
be tailored to suit the needs of the experiment. These can be
bilayers [1–5], self-assembled monolayers [6, 7], or tethered
bilayers, using the integration of other molecules as supports distancing the layer from the surface [8, 9]. The adaptability of the
approach has led to exciting new developments in
bio-nanotechnology [10, 11] and in the design of novel characterization methods for investigating molecular interactions, membrane
protein–lipid interactions, and antimicrobial peptides [3, 12–
14]. For the study of protein–ligand interactions, the layers can
be adapted with the addition of lipids with functional head groups
Tina Daviter et al. (eds.), Protein-Ligand Interactions: Methods and Applications, Methods in Molecular Biology, vol. 2263,
https://doi.org/10.1007/978-1-0716-1197-5_7, © Springer Science+Business Media, LLC, part of Springer Nature 2021
183