Simultaneous nanoplasmonic and quartz crystal microbalance sensing: analysis of biomolecular conformational changes and quantification of the bound molecular mass.
This paper presents a study of supported lipid bilayer (SLB) formation and subsequent protein binding using a sensor that combines localized surface plasmon resonance (LSPR) and quartz crystal microbalance with dissipation (QCM-D) monitoring. The LSPR activity arises from silicon oxide (SiO x ) coated nanometric apertures in a thin gold film, which also serves as the active electrode of a QCM-D crystal. Both transducer principles provide signatures for the formation of a SLB upon adsorption and subsequent rupture of adsorbed lipid vesicles. However, the two techniques are sensitive over different regions of the sample: LSPR primarily inside and on the rim of the holes and QCM-D primarily on the planar areas between the holes. Although the dimension of the lipid vesicles is on the same order as the dimension of the nanoholes, it is concluded from the response of the combined system that vesicle rupture in the nanoholes and on the planar region between the holes is synchronized. Furthermore, by determining the thickness of the SLB from the QCM-D response, the characteristic decay length of the LSPR field intensity could be determined. This made it possible not only to determine the mass and refractive index of the homogeneous SLB but also to postulate a generic means to quantify the LSPR response in terms of mass-uptake also for nonhomogeneous films. This is exemplified by measuring the adsorbed lipid mass during vesicle adsorption, yielding the critical lipid vesicle coverage at which spontaneous rupture into a planar bilayer occurs. The generic applicability and versatility of the method is demonstrated from specific protein binding to a functionalized SLB. From the absolute refractive index of the protein, provided from the LSPR data alone, it was possible to determine both the effective thickness of the protein film and the molecular mass (or number) of bound protein.
Division of Solid State Physics, Lund University, SE-22100 Lund, Sweden. magnus.p.jonsson@chalmers.se
Magnus P
MP
Jonsson
Peter
P
Jönsson
Fredrik
F
Höök
D015374:Biosensing Techniques / Q000295:instrumentation* / Q000379:methods*; D003460:Crystallization; D004566:Electrodes; D008968:Molecular Conformation; D008970:Molecular Weight; D049329:Nanostructures / Q000737:chemistry*; D011791:Quartz / Q000737:chemistry*
D011791:Quartz