FG Nanobiotechnologie
Routine Surface Plasmon Resonance (SPR) measurements, which are widely applied in affinity (bio)sensors and surface science, do not allow one to separate the obtained SPR signals into the change of the refractive index and thickness of deposited layers. For thin layers the signal is proportional to the product of the layer thickness and the difference in the refractive indices of this layer and of the aqueous media. In this study, we suggest an approach to separate these parameters. It is performed by subsequent measurements in the presence and in the absence of polyethylene glycol (PEG), an inert additive which modifies the refractive index of the liquid phase but do not penetrate into the adsorbed layer. Both parameters were successively determined for human serum albumin (HSA) as well as for anti-HSA IgG in three different settings: i) HSA adsorbed directly onto a gold surface, ii) HSA chemically immobilized on a gold surface coated with a self-assembled monolayer of 1,16 mercaptohexadecanoic acid and iii) chemically immobilized HSA with additional monomolecular layer of anti- HSA IgG antibodies bond to this protein layer. The suggested approach can be applied in most SPR devices with a flow cell for analysis of various adsorbed layers.
Despite the history of application of surface plasmon resonance (SPR) for chemo- and biosensing being over 30 years long, the development of this technique is still in progress.
This review is focused on the technological aspects of further improvement of analytical performance of SPR transducers based on Kretschmann configuration. We describe basic measurement configurations, their improvements and optimizations, and their drawbacks and limitations. An importance of referencing in SPR sensors is highlighted. The referencing approaches are classified into the following domains: (1) macroscopic spatially separated referencing, (2) self-referencing based on micro-patterning, (3) in-place referencing, (4) spatiotemporal referencing, and (5) electrochemically assisted referencing. The underlying principles of these approaches, examples of their implementation, and resulting improvements of sensor performance are described. Finally, an analysis of SPR data and an extraction of affinity properties are discussed.
SPR based biosensors register binding of analytes to the surface with immobilized receptors by measuring changes of the refractive index near this surface. An important task in the improvement of this measurement technology is a separation of signals, corresponding to the changes in the chemosensitive layer, from undesired contributions of bulk phase, for example, due to fluctuations of temperature, concentrations of solutes, pressure. The wavelength of the incident light influences strongly the penetration depth of the corresponding evanescent wave. This dependence was exploited here for compensation of the contribution of the bulk refractive index. It was performed using differential SPR measurements at two wavelengths with differing penetration depths. Theoretical analysis and numerical optimization of the suggested approach, named a Penetration Difference Self-Referencing SPR (PDSR-SPR), were performed. Experimental test was performed using 658 and 980 nm laser diodes. Over 20 times suppression of variations of bulk refractive index with magnitude up to 1000 μRIU was observed. Finally, PDSR-SPR approach was applied for monitoring of antibodies binding to the immobilized antigens.
Mercury reduction and oxidation on ultrathin gold electrodes was investigated by simultaneous voltammetry and surface plasmon resonance (SPR). The angle dependencies of SPR signal were analyzed quantitatively. The results indicate partially reversible formation of amalgam with optical properties in between of optical properties of mercury and gold. Up to 12% content of mercury in the gold layer is suggested.
A DNA-based surface plasmon resonance biosensor for enrofloxacin was developed. Heating denatured DNA immobilized on the gold-coated glass surface was exploited. The immobilization was performed by a layer-by-layer co-deposition with a cationic polymer. The sensor performance was tested with real biological probes. Direct and simple determination of enrofloxacin in milk samples was demonstrated. The sensor response obeys Langmuir binding isotherm being almost linear until about 20 µg per mL. The detection limit in milk samples was estimated to be 3 µg per mL.
The optical properties of electrochemically polymerized N-methylaniline are changed in the presence of gaseous hydrogen chloride. This effect was used for preparation of chemical sensors with transduction based on surface plasmon resonance. The interaction of hydrogen chloride with a two nm layer of poly-(N-methylaniline) on gold leads to the shift of the surface plasmon resonance. The analysis of the resonance spectra demonstrates that the effect is caused by an increase of the imaginary component of the refractive index; a minor contribution is also provided by film condensation leading to an increase of the real component of the refractive index and a decrease in the thickness. The effect is selective and quasi-reversible. The concentration dependence of the gas effect obeys Langmuir's adsorption isotherm with a reciprocal value of the binding constant of 850 ± 160 ppm.
Chiroselective binding sites have been created on thin gold films by application of the spreader-bar approach.
Impedometric techniques and surface plasmon resonance were applied to detect binding. (R)-(+)-1,1 '-Binaphthyl-2,2 '-diol (R-BNOH) and (S)-(-)-1,1
'-binaphthyl-2,2'-diol (S-BNOH) were used as model analytes. The artificial receptors were prepared by co-adsorption of 16-mercaptohexadecane (matrix) with a thiol-modified chiral selector (template). The conjugates of d, l-thioctic acid and (R)-(+)- or (S)-(-)-1,1'-binaphthyl-2,2'-diamine were used as templates. Different concentration ratios of the matrix and template were tested. No chiral selectivity of surfaces formed by either the matrix or the template alone was observed. The use of alkylthiols shorter than 16-mercaptohexadecane led to the formation of surfaces with no chiral selectivity. The gold electrodes coated by the spreader-bar technique displayed an enantioselectivity of up to 4.76 or up to 2.55 as measured by the capacitive and SPR methods, espectively.
Bimetallic layers increase sensitivity of affinity sensors based on surface plasmon resonance
(2002)
Two metals are used in resonant layers for chemical sensors based on surface plasmon resonance (SPR) - gold and silver. Gold displays higher shift of the resonance angle to changes of ambient refraction index and is chemically stable. Silver posses narrower resonance curve thus providing a higher signal/noise ratio of SPR chemical sensors, but has a poor chemical stability. A new structure of resonant metallic film based on bimetallic silver/gold layers (gold as an outer layer) is suggested. It combines advantages of both gold and silver resonant layers. Bimetallic resonant films display so high shift of resonance angle on changes of ambient refraction index as gold films, but show narrower resonance curve, thus providing a higher signal / noise ratio. Additionally, the outer gold layer protects silver against oxidation.