Human immunoglobulin adsorption investigated by means of quartz crystal microbalance dissipation, atomic force microscopy, surface acoustic wave, and surface plasmon resonance techniques.

Time-resolved adsorption behavior of a human immunoglobin G (hIgG) protein on a hydrophobized gold surface is investigated using multitechniques: quartz crystal microbalance/dissipation (QCM-D) technique; combined surface plasmon resonance (SPR) and Love mode surface acoustic wave (SAW) technique; combined QCM-D and atomic force microscopy (AFM) technique. The adsorbed hIgG forms interfacial structures varying in organization from a submonolayer to a multilayer. An "end-on" IgG orientation in the monolayer film, associated with the surface coverage results, does not corroborate with the effective protein thickness determined from SPR/SAW measurements. This inconsistence is interpreted by a deformation effect induced by conformation change. This conformation change is confirmed by QCM-D measurement. Combined SPR/SAW measurements suggest that the adsorbed protein barely contains water after extended contact with the hydrophobic surface. This limited interfacial hydration also contributed to a continuous conformation change in the adsorbed protein layer. The viscoelastic variation associated with interfacial conformation changes induces about 1.5 times overestimation of the mass uptake in the QCM-D measurements. The merit of combined multitechnique measurements is demonstrated.

Biosensing based on light absorption of nanoscaled gold and silver particles.

The absorption spectrum of noble metal spherical nanoparticles is known to be strongly influenced by the dielectric constant of the surrounding material, and as such, these particles are well suited for biosensing applications. To perform biosensing using nanoparticles on a substrate, the metal particles are covalently attached onto quartz using an organic adhesion layer of mercaptosilanes. The particles in solution are characterized by UV-vis spectroscopy and transmission electron microscopy, while those attached to the quartz are characterized with UV-vis spectroscopy and atomic force microscopy. Antibodies are attached to the metal nanoparticles, and the antigen recognition is monitored via the change of light absorption when this binding event occurs. Not only is the absorbance originating from plasmon resonances of the particles influenced by the dielectric properties of molecules attached to the nanospheres but also the interband absorption of the particles changes, which will be demonstrated in this report. A light absorption change is detected when a molecular recognition occurs between the bioreceptor molecules attached to the nanoparticle and a biomolecular counterpart. This change in absorption can be very large when adhered molecules are at resonance (interband transitions). In addition, the presented type of biosensing can be a cost-effective and easy to use alternative to conventional biosensing techniques.