Simultaneous Surface Plasmon Resonance and Quartz Crystal Microbalance with Dissipation Monitoring Measurements of Biomolecular Adsorption Events Involving Structural Transformations and Variations in Coupled Water

摘要

Simultaneous quartz crystal microbalance with dissipation monitoring (QCM-D) and surface plasmon resonance (SPR) measurements are used to analyze the surface kinetics of two biomacromolecular systems, one lipid and one protein based, undergoing surface-induced conformational changes. First we establish a theoretical platform, which allows quantitative analysis of the combined SPR and QCM-D data. With this theoretical base, new information can be extracted, not obtainable with either technique alone. As an example we demonstrate how time-resolved measurements with these two techniques in combination-yielding three independent measured quantities-add new information about (i) kinetics, i.e. number of adsorbed molecules per unit area versus time, and (ii) temporal variation in the mass fraction of coupled water versus coverage. In particular, it is demonstrated for the first time, how the kinetics of the process during which adsorbed vesicles are spontaneously transformed into a supported phospholipid bilayer (SPB) on SiO_(2) can be quantitatively separated into its two dominating states: adsorbed vesicles and supported planar bilayer patches. In addition, the relevance of dynamically coupled water for interpretation and modeling of the QCM-D response during bilayer formation is discussed and further illustrated with a second model system: streptavidin adsorption on a biotin-modified SPB. A very strong coverage dependence in the number of water molecules per protein sensed by the QCM is demonstrated, with strong implications for the use of QCM as a tool for quantitative determination of protein mass uptake kinetics.