Surface plasmon resonance imaging (SPRi) as an alternative technique for rapid and quantitative screening of small molecules, useful in drug discovery

Surface plasmon resonance imaging (SPRi) is a label free technology for biomolecular interaction, which gives access to binding kinetic parameters from real time acquisition. It offers the possibility to test in a single run a large number of interactions, allowing rapid identification of the most suitable compounds toward a given biological entity. Until now, this technique has proven to be relevant for interaction between relatively large molecules (protein, antibodies, DNA) but has not been challenged yet for the screening of small molecules that can be of interested in the field of drug discovery. As a proof a principle, we have used SPRi to screen for interaction of several small molecules, referred to as G4-ligands on G-quadruplex DNA. This technology allowed to easy discrimination of the binding properties of four G4-ligands on quadruplex DNA models.     

Phase-sensitive spatially-modulated surface plasmon resonance polarimetry for detection of biomolecular interactions

We here describe a scheme of spatially modulated surface plasmon resonance (SPR) polarimetry that enables to combine ultra-high phase sensitivity with good signal-to-noise background. The proposed approach uses spatial modulation of s-polarized component by birefringent elements and the extraction of phase-polarization information by Fourier-transform methods. This scheme was tested for monitoring the interactions between an antibody and its biological partner. Our experimental data, collected by amplitude-sensitive and phase-sensitive polarimetry demonstrate that the latter scheme provides at least one order of magnitude improvement in terms of detection limit.     

Sensitivity Comparison of Surface Plasmon Resonance and Plasmon-Waveguide Resonance Biosensors

Plasmon-waveguide resonance (PWR) sensors are particularly useful for the investigation of biomolecular interactions with or within lipid bilayer membranes. Many studies demonstrated their ability to provide unique qualitative information, but the evaluation of their sensitivity as compared to other surface plasmon resonance (SPR) sensors has not been broadly investigated. We report here a comprehensive sensitivity comparison of SPR and PWR biosensors for the p-polarized light component. The sensitivity of five different biosensor designs to changes in refractive index, thickness and mass are determined and discussed. Although numerical simulations show an increase of the electric field intensity by 30-35% and the penetration depth by four times in PWR, the waveguide-based method is 0.5-8-fold less sensitive than conventional SPR in all considered analytical parameters. The experimental results also suggest that the increase in the penetration depth in PWR is made at the expense of the surface sensitivity. The physical and structural reasons for PWR sensor limitations are discussed and a general viewpoint for designing more efficient SPR sensors based on dielectric slab waveguides is provided.     

Comparison of surface plasmon resonance spectroscopy and quartz crystal microbalance for human IgE quantification

Surface plasmon resonance (SPR) and quartz crystal microbalance (QCM) have been known independently as surface sensitive analytical devices capable of label-free and in situ bioassays. In this study a SPR device and a 10 MHz QCM sensor are employed for the study of human IgE and anti-human IgE-binding reactions upon immobilizing the latter on the gold electrodes. The SPR and QCM response curves to the antibody immobilization and antigen binding are similar in shape but different in time scale, reflecting different resonation principles. Through optimization of the anti-human IgE coating, both the SPR and QCM sensors could detect IgE in a linear range from 5 to 300 IU/ml. Although the intrinsic sensitivity of the SPR device is five times of the 10 MHz QCM, the IgE detection sensitivity of the two methods is, however, different only in a factor of 2. The acceptable QCM sensitivity for the IgE detection is attributed to the fact that QCM measures the sum of molar mass of a protein layer and the entrapped water. Although both the devices use open, stand still liquid cell, and all the measurements are performed at room temperature, the SPR reproducibility and reliability are better than QCM, as the QCM frequency is more sensitive to temperature fluctuations, press changes and mechanical disturbances.     

Highly accurate and sensitive surface plasmon resonance sensor based on channel photonic crystal waveguides

A highly sensitive surface plasmon resonance (SPR) sensor based on channel photonic crystal waveguide (PCW) is proposed. The PCW is based on widely used lithographic and nano-fabrication compatible materials like TiO₂ and SiO₂. Gold has been used as a SPR active metal. By rigorously optimizing the different waveguide parameters, we have shown that there is significant transfer of modal power around phase-matching or resonance wavelength which has been utilized to design a compact and highly sensitive sensor for lab on chip. The ultra narrow width (∼765 pm for an interaction length of 10 mm) of surface plasmon resonance curve and sensitivity as high as 7500 nm-RIU⁻¹ will open a new window for bio-chemical sensing applications.     

Full-field surface plasmon resonance sensor for high resolution refractive index measurement using common-path heterodyne interferometer

This study presents a full-field surface plasmon resonance (SPR) sensor induced by attenuated total reflection (ATR)-couple for liquid refractive index measurement. The system adopts a common-path heterodyne interferometer to measure the phase difference between P- and S-wave after passing through the SPR sensor. In order to realize the full-field measurement, it adopts a three-frame integrating-bucket method. The experimental results show great consistency profile between single point and full-field liquid refractive index measurement from 1.330 to 1.340 RIU. It shows that the best sensitivity and resolution of a single pixel in charge couple device (CCD) for liquid refractive measurement are 3.3 × 10⁴ (deg/RIU) and 3.53 × 10⁻⁶ (RIU), respectively. As compared with traditional single-point method, the proposed method with a regular CCD has no degradation. Therefore, the system has many applications in chemistry and biology.     

Searching for optimal sensitivity of single-mode D-type optical fiber sensor in the phase measurement

To improve the sensitivity of a single-mode D-type optical fiber sensor, we selected a D-type optical fiber sensor with 4 mm long and 4 μm core thickness made of a single-mode fiber, a Au-coating on the sensor with a thickness range of 15–32 nm, a light wavelength of 632.8 nm, and an incident angle of 86.5–89.5° for different refractive index (1.33–1.40) sensing. These simulations are based on the surface plasmon resonance (SPR) theory using the phase method which shows that the sensitivity is proportional to the refractive index, Au film thickness and lower incident angle on the sensing interface. The sensitivity is higher than 4000 (degree/RIU), and the resolution is better than 2.5 × 10⁻⁶(RIU) as the minimum phase variation is 0.01°. This device is used to detect the refractive index or gas or liquid concentration in real-time. The proposed sensor is small, simple, inexpensive, and provides an in vivo test.     

Utilization of albumin-based sensor chips for the detection of metal content and characterization of metal–protein interaction by surface plasmon resonance

We report here the use of albumin-based biosensor chips for the determination of metal content and characterization of metal–protein interaction by surface plasmon resonance. Bovine serum albumin was immobilized onto a carboxymethylated dextran matrix and used for metal detection. The temperature for the analysis was defined and the highest interaction was observed at 25 °C. The albumin sensor chip binds cadmium, zinc or nickel in a concentration-dependent manner, but not magnesium, manganese and calcium. The optimal buffer condition used for the analysis contains 0.01 M HEPES, pH 7.4, 1 mM NaCl and 0.005% Tween-20. Using this condition, a linear calibration curve within the range of 10⁻⁸ to 10⁻⁴ M can be established for the metals. However, a dramatic increase in binding capacity was observed when metal concentration was higher than 10⁻⁴ M and reached a plateau at 10⁻² M. The detection limit for Cd can reach as low as 1 ppb. When measuring a solution containing two species of metal ions with the albumin chip, an additive effect was observed for Ni and Zn. However, 20–30% reduction in resonance response was found upon mixing Cd with Zn or Ni. These observations are consistent with the binding characteristics of albumin. The feasibility of measuring serum metal content by the albumin chip was examined. A linear calibration curve can be established if the samples are boiled and passed through a gel filtration column. The binding affinity of metal with albumin can also be achieved by using the sensor chip. The binding affinity follows the order of Ni > Zn > Cd. These results indicate that the albumin-based sensor chip is useful not only in the quantification of metal content, but also in the characterization of the biochemical properties of albumin.     

Manufacture of robust surface plasmon resonance fiber optic based dip-probes

Surface plasmon resonance (SPR) spectroscopy has been conducted on both prism and fiber optic (FO) based sensors for several years. This technique measures the refractive index (RI) of a solution or layer adsorbed to a thin (50 nm) Au layer on the sensor substrate. To date a succinct set of protocols have not been published regarding the optimization of fiber-based SPR dip-probe sensors. Such sensors would allow application of SPR to a wider variety of applications. This paper focuses on consideration of the choice of fiber, isolation of the mirror from the sensing area, and orientation of the probes in the metal layer sputter deposition chamber in the manufacture of SPR dip-probes for reproducibility and robustness. Optimization of the process yields sensors with a batch to batch reproducibility as low as 0.5 nm in the location of the SPR spectral minima. Further study of RI measurements by the same probe over 2 months show these SPR dip-probes have a long shelf-life. A selection of probes was exposed to various solutions to monitor their drift. The data shows the probes’ response indicated a lowering of the RI measured over a period of 3 or 7 days depending on the probe type. Evidence of surface porosity and damage upon exposure to hydrothermal water seems to indicate these sensors are prone to chemical attack. Further research is needed to characterize this attack and allow creation of more robust sensors.     

Surface plasmon resonance detection of small molecule using split aptamer fragments

It was difficult to detect small molecules directly using conventional surface plasmon resonance (SPR) biosensors since the changes of refractive index, which was resulted by binding small molecules, were usually small. In this paper, split aptamer fragments were used for the construction of SPR biosensor to determine small molecule such as adenosine with high sensitivity. An aptamer for adenosine was designed to be two flexible ssDNA pieces, one was tethered on Au film and the other was modified on Au nanoparticles (AuNPs). In the presence of adenosine, two ssDNA pieces reassembled into the intact aptamer structure and the AuNPs-labeled adenosine-aptamer complex was formed on the Au film. Then, the resonance wavelength shift was enhanced obviously, due to the electronic coupling between the localized plasmon of AuNPs and the surface plasmon wave associated with Au film. The results confirmed that this biosensor could detect adenosine with high sensitivity and selectivity. The limitation of detection (LOD) of this SPR biosensor was ca. 1.5 pM, which was an approximately ca. 2-3 order of magnitude lower than that of those SPR biosensors which utilized competitive methods.     

Surface plasmon resonance based fiber optic sensor for the detection of low water content in ethanol

A fiber optic sensor utilizing surface plasmon resonance (SPR) has been fabricated for the detection of low content of water in ethanol. The sensor utilizes spectral interrogation technique for operation. The resonance wavelength has been found to vary linearly with water content in the range 0-10% with sensitivity of 1.149 nm per percentage of water. The results are in agreement with the refractive index variation of ethanol-water mixture. The sensor has a water resolution of 0.145% which is better than the evanescent wave absorption sensor reported for the similar study. The sensor will find application in determining the low water content in ethanol which is used as a bio-fuel and in the field of medicine and organic chemistry.     

A spectral imaging array biosensor and its application in detection of leukemia cell

A new type of array immunosensor was developed by combining surface plasmon resonance (SPR) and spectral imaging techniques. The system consisted of a monochromator as the wavelength scanning light source, a polarizer, Kretschmann-Raaether attenuated total reflection (ATR) configuration including array sensor chip, and a CCD camera. The images of transmitting light from ATR were recorded versus the wavelength. By averaging gray scales of the pixels in the area of every gold spot from the image series, the complete spectral resonance curve of all sensing spots on the array can be extracted in parallel. The performance of the developed system was evaluated by analyzing interactions of the anti-CD33 monoclonal antibody to its target leukemic cells using 11 cases of human bone marrow specimens. The specimens were also analyzed with flow cytometry method (FCM) for comparison. The initial results measured by the immunosensor array were corresponded with that of FCM, indicating that the developed parallel method might be clinically suitable for immunophenotyping of acute leukemias. The new sensor array system showed the merits of high-throughput, high sensitivity, high specificity, label free and operation convenient. Spots numbers of the array could be increased if suitable technology were adopted for manipulating the micro bio-liquids on the sensor array chip.     

A Numerical Approach to Design the Kretschmann Configuration Based Refractive Index Graphene-MoS2 Hybrid Layers With TiO2-SiO2 Nano for Formalin Detection

In this paper,a Kretschmann configuration based surface plasmon resonance (SPR) sensor is numerically designed using graphene-MoS2 hybrid structure TiO2-SiO2 na...     

Surface functionalization for self-referencing surface plasmon resonance (SPR) biosensors by multi-step self-assembly

Recently, a novel SPR sensor with on-chip referencing has been realized. In this sensor, one-half of the gold sensing surface is covered with a high refractive index overlayer of tantalum pentoxide (Ta₂O₅). When polychromatic beam illuminates the sensing surface, surface plasmon resonance in the areas with and without the overlayer occur at different wavelengths. Therefore, the reflected light exhibits two dips associated with SPRs in those two areas. When functionalized properly, one of the areas can be used as a specific sensing channel for detection of specific bio-interactions and the other can act as a reference channel for compensation for background refractive index fluctuations. In this paper we present a new functionalization approach for these mixed architecture chips. The gold side of the chip is functionalized with a mixed self-assembled monolayer of polyethylene oxide (PEO) and biotin terminated (BAT) thiols whereas the Ta₂O₅ side is coated with PEO terminated silanes. The PEO terminated thiols and silanes serve as a protein resistant background, while the biotin terminated thiols are used to bind streptavidin, which in turn immobilizes biotinylated antibodies. Hence, the gold side of the chip is used for the binding and detection of target analytes and the Ta₂O₅ side functions as a reference channel that monitors bulk refractive index changes and temperature drift. We have studied human chorionic gonadotropin (hCG) as a model system, currently detecting down to 5 ng/ml. In addition, we demonstrate the power of the on-chip reference channel for compensating for refractive index changes and eliminating false alarms.     

3D-quantification of biomolecular covers using surface plasmon-polariton resonance experiment

The concentration of surface molecules N_s and components of molecular susceptibility χ_jl(ω) can both be determined from surface plasmon-polariton resonance (SPPR) experiments, instead of effective layer thickness and index of refraction, which are usually determined. The theoretical consideration of a molecular layer as monolayer of separated 3D-oscillators provides a new perspective for investigating molecules during SPPR experiment. It is shown that SPPR response and the form of the reflective curve depend on the form of a biomolecule and its orientation relative to the surface of the metal-carrier of plasmon oscillations. The experimental data for immunological reaction for the calculation of surface molecular concentration and mass of biomolecular covering are presented.     

Signal enhancement of surface plasmon resonance-based immunoassays for the allergen detection

A surface plasmon resonance (SPR) biosensor was used to determine the recombinant group 1 house dust mite allergen (rDer f1) in both HBS-EP buffer and fetal bovine serum (FBS). The monoclonal antibody was immobilized onto the CM5 sensor chip surface using an amine coupling method. The procedures of antibody immobilization and the subsequent primary and enhanced immunoassay were monitored in real time. The sensitivity for rDer f1 detection was remarkably improved by using intact polyclonal antibody as signal amplifying agent. Using this signal enhanced SPR immunosensor, rDer f1 in HBS-EP buffer and FBS was detected at a concentration of 15.4 and 32.1 ng/ml, respectively. The result demonstrates that SPR biosensor is a simple and reliable method for allergen detection.     

Sensitivity optimization of surface plasmon sensors for detection of intermediate layers

A sensitivity optimization method for the detection of an intermediate layer using a surface plasmon sensor is presented. The dependence of detection sensitivity on distance from the metal surface and the average refractive index over the sensing region are considered. Based on the calculated results, the sensitivity for detecting an intermediate layer in a multilayered sample is determined by varying the refractive index and thickness of each layer. It is shown that, in particular cases, controlling the refractive index and thickness increases the detection sensitivity. The proposed method is useful for designing multilayered samples.     

Sensitivity of surface plasmon resonance sensors based on metallic columnar thin films in the spectral and angular interrogations

Surface Plasmon Resonance (SPR) from metallic Columnar Thin Films (CTFs) of porosity as high as 0.5 was experimentally and theoretically investigated. The CTF layers were prepared by the Glancing Angle Deposition (GLAD) method. The SPR features were investigated in both the angular and the spectral modes. In the angular interrogation, increasing the porosity causes broadening to the dip width, shift to larger resonance angles, and increase of the sensitivity to analyte refractive index (RI) changes by about threefold compared with closed metal films. In the spectral interrogation, on the other hand, the resonance wavelengths are red-shifted for porous films; hence their spectral sensitivities are higher than those of closed films under the same experimental conditions. Nevertheless, the sensitivity behavior versus the resonance wavelength is similar to that of SPR sensors based on dense film layers. The shapes of the nanostructures constituting the CTF are described as ellipsoidal inclusions in which the effective permittivity dyadic of the composite material is calculated using the Bruggeman formalism with exact depolarization dyadics. The correlation between the sensitivity enhancement and the electromagnetic field intensity at the interface between the metallic film and the analyte was examined. Electromagnetic fields analyses were performed using the general 4 × 4 propagation matrices of general homogenous biaxial layers.     

An application of a plasmonic chip with enhanced fluorescence to a simple biosensor with extended dynamic range

Plasmonic chips coated with thin silver and SiO₂ layers were applied to a simple biosensor with wide dynamic range, which has some advantages in the points of sensitivity and surface selectivity in spite of a simple fluorescence detection system without complex optical stuffs. The enhanced fluorescence of fluorescent labeled-marker proteins, cy5-streptavidin (SA), excited by irradiation light from the rear panel of the chip was measured. To obtain the maximum fluorescence enhancement by excitation under an electric field enhanced by grating-coupled surface plasmon resonance from the rear irradiation, the optimal silver thickness was studied and found to be 30-35 nm. A directional fluorescence due to re-coupling between fluorescence and plasmon polaritons was also observed on the plasmonic chips and enhanced fluorescence 90 times compared with that on glass slides was superior to the others reported. The limit of detection (LOD) studied here was improved up to 50 pM after 10-min incubation and dynamic range was also extended by 2 order, compared with that on glass slides. The irradiation from the rear panel demonstrated advantage in the surface-selective detection of cy5-SA at 100 nM concentration. Our plasmonic chip has great potential in biosensing application due to its advantages of rapid detection, simple operation, high sensitivity (wide dynamic range), surface selectivity, and suppression of non-specific adsorption.