Surface Plasmon Resonance Signal Enhancement Research Articles

Cucurbit[7]urils induced bimetallic nanoparticles network for ultra-sensitive detection of Caspase-3 based on surface plasmon resonance

A novel signal amplification method for surface plasmon resonance (SPR) was developed to detect Caspase-3 based on cucurbit[7]urils (CB[7]) and bimetallic nanonetwork. Three-dimensional (3D) bimetallic nanonetwork was formed through CB[7]-integrated self-assembly of gold nanoparticles (AuNPs) and silver nanoparticles (AgNPs) (AuNPs-CB[7]-AgNPs) by one-pot synthesis method. Caspase-3 was utilized as a model protein and peptide EEAAADEVDFKKAAAC was used as the substrate. In the presence of Caspase-3, peptide was hydrolyzed and exposed phenylalanine at the N-terminus, which could bind to CB[7] through host-guest interaction. The multivalent receptors CB[7] on bimetallic nanonetwork provide a simple strategy to increase the binding affinities between 3D bimetallic nanonetwork and peptide. Thus, AuNPs-CB[7]-AgNPs could attach to the gold chip with ease. Remarkably, due to the strong optical properties and high refractive index of 3D bimetallic nanonetwork, it has a higher SPR signal enhancement than that of only AuNPs or AgNPs. Therefore, a highly sensitive Caspase-3 sensing strategy could be achieved with a low detection limit of 5.39 pg mL−1, providing a new and facile SPR signal amplification method for detection of cell apoptosis.

Surface plasmon resonance signal enhancement based on erlotinib loaded magnetic nanoparticles for evaluation of its interaction with human lung cancer cells

Surface plasmon resonance (SPR) sensor provides a very useful tool based on its label-free, real-time monitoring and low price properties. However, measurement of small molecules and extremely diluted analytes is difficult and therefore, signal enhancement is required. In the present study, signal enhancement of erlotinib conjugated magnetic nanoparticles (erlotinib-MNPs) compared to erlotinib was evaluated via their interaction with overexpressed epidermal growth factor receptor on human lung cancer cells (A549 cell line) surface using SPR sensor at three temperature levels. The attained results showed an average signal amplification of about 2.5-fold for MNP-erlotinib interaction with A549 cells compared to erlotinib interaction with those cells. Overall, this signal enhancement method could be used for measurement of small molecules and cancer cell detection via interaction of their overexpressed receptor and various ligands.

Application of Gold and Platinum Bimetallic Nanoparticle to enhance Surface Plasmon Resonance Signal

Sensors based on the surface plasmon resonance (SPR) technique are useful devices to detect and monitor interactions between biomolecules in real-time. SPR is a label-free method that monitors the variation of reflectivity of a biochip composed of a metal-coated glass prism and can be applied in several areas, such as biotechnology, food safety and clinical diagnosis. In the last years, several researchers have proven the efficiency of metallic nanoparticles (NPs) in the enhancement of SPR signal. This feature allowed the detection of biomolecules at very low concentration. Aiming to further enhance SPR signal towards the detection of proteins at low concentration and by a simple procedure, the present work compared the performance of gold and platinum bimetallic NPs (AuPtNPs) with that of monometallic gold NPs (AuNPs) in the enhancement of SPR signal. In order to evaluate the NPs, protein peanut agglutin (PNA) was used as target analyte and anti-PNA antibody was used as sensing molecule. Firstly NPs were functionalized with anti-PNA antibody and incubated with a solution containing PNA. Then, the NPs bound to PNA were injected into the SPR equipment containing a biochip previously modified with anti-PNA antibody. The results demonstrated that the AuPtNPs provided a 91-fold increase compared to the direct detection of free PNA in solution. In comparison with AuNPs, the signal generated by AuPtNPs was about 4 times higher. This encouraging result indicated that the application of bimetallic NPs may be a better strategy to further enhance sensitivity of SPR biosensors and could drive the development of new strategies that are not only simple, but also able to detect proteins at low concentrations, which is of great importance, especially in clinical diagnostics.

Gold nanoprism/Tollens’ reagent complex as plasmonic sensor in headspace single-drop microextraction for colorimetric detection of formaldehyde in food samples using smartphone readout

In this work, an assay with high sensitivity and selectivity for the detection of formaldehyde (FA) is presented. The assay applied a gold nanoprism/Tollens’ reagent (Au-np/TR) complex as the sensor used in headspace single-drop microextraction (HS-SDME). A surface plasmon resonance signal enhancement as well as color change was caused by the formation of Au@Ag-np after a redox reaction between FA and TR during the HS-SDME process. With the utilization of smartphone nanocolorimetry (SNC), the FA could be detected and quantified. For HS-SDME-SNC, a linearity calibration curve ranging from 0.1 to 100 μM was obtained, and the limit of detection was determined to be 30 nM. Successful attempts to determine FA were demonstrated by analysis of the analyte in (adulterated) raw food samples (octopus and chicken flesh). Matrix effects from real samples were avoided by using HS-SDME, and only a 3-μL droplet of solvent was needed in the assay.

Stimuli-Responsive Microgel-Based Surface Plasmon Resonance Transducer for Glucose Detection Using a Competitive Assay with Concanavalin A

Glucose responsive microgels composed of poly(N-isopropylacrylamide-co-glycosyloxyethyl methacrylate) (p(NIPAm-co-GEMA)) were synthesized by free radical precipitation polymerization. The resultant microgels were shown to collapse upon addition of Concanavalin A (ConA) owing to its ability to bind up to four GEMA molecules. We showed that the microgel could reswell upon addition of free glucose due to the competitive binding of GEMA and glucose with ConA, effectively breaking the cross-links responsible for the collapse. Thus, by monitoring the contraction/expansion process of the p(NIPAm-co-GEMA) microgels bound to a Au surface via surface plasmon resonance (SPR), the concentration of glucose in solution can be quantified. We go on to show that there is a ninefold SPR signal enhancement compared to exposure of bare Au to glucose and a similar enhancement compared to microgels without GEMA. Finally, we showed that the p(NIPAm-co-GEMA) microgel-based SPR sensor surfaces were not responsive to other interfering species. Taken together, the results show that the competitive binding concept can be useful for detecting small biologically relevant molecules that are usually difficult to detect via SPR. Therefore, this concept can be expanded upon to detect other small molecules of interest in the future.

SPR signals enhancement by gold nanorods for cell surface marker detection.

Introduction: The detection of micrometer-sized particles like cells is limited by surface plasmon resonance (SPR) biosensors because of having a depth of evanescent wave <500 nm. In this study, for the first time, we exhibited the use of streptavidin-functionalized gold nanorods (GNRs) as intensification labels for detection of cell surface markers in SPR-based biosensors. Methods: The GNRs (ʎ max: 735 nm) were modified with streptavidin using EDC/NHS coupling method and human umbilical vein endothelial cells (HUVECs) were selected as the cell model for detecting VE-cadherin on cell surface using real-time SPR device in the 785 nm wavelength of the laser source. Results: The investigations revealed that the plasmonic field extension produced from the gold layer and GNRs resulted in multiple enhancement of SPR signals when the wavelength of laser source in SPR instrument was matched with the wavelength of maximum absorbance in GNRs. Moreover, the results showed that the growth of ∆RU value in specific and non-specific bindings for various cell number injections were produced with increasing the cell number. Conclusion: The results displayed that cell detection can be performed in real- time form without any need to a time-consuming process used in conventional methods like immunocytochemistry, flow cytometry, and western blotting.

Gold Nanoparticles Used as Protein Scavengers Enhance Surface Plasmon Resonance Signal.

Although several researchers had reported on methodologies for surface plasmon resonance (SPR) signal amplification based on the use of nanoparticles (NPs), the majority addressed the sandwich technique and low protein concentration. In this work, a different approach for SPR signal enhancement based on the use of gold NPs was evaluated. The method was used in the detection of two lectins, peanut agglutinin (PNA) and concanavalin A (ConA). Gold NPs were functionalized with antibodies anti-PNA and anti-ConA, and these NPs were used as protein scavengers in a solution. After being incubated with solutions of PNA or ConA, the gold NPs coupled with the collected lectins were injected on the sensor containing the immobilized antibodies. The signal amplification provided by this method was compared to the signal amplification provided by the direct coupling of PNA and ConA to gold NPs. Furthermore, both methods, direct coupling and gold NPs as protein scavengers, were compared to the direct detection of PNA and ConA in solution. Compared to the analysis of free protein, the direct coupling of PNA and ConA to gold NPs resulted in a signal amplification of 10–40-fold and a 13-fold decrease of the limit of detection (LOD), whereas the use of gold NPs as protein scavengers resulted in an SPR signal 40–50-times higher and an LOD 64-times lower.

Quantum dot monolayer for surface plasmon resonance signal enhancement and DNA hybridization detection

We report results of studies relating to the fabrication of a surface plasmon resonance (SPR)-based nucleic acid sensor for quantification of DNA sequence specific to chronic myelogeneous leukemia (CML). The SPR disk surface has been modified with octadecanethiol self-assembled monolayer followed by deposition of the tri-n-octylphosphine oxide capped cadmium selenide quantum dots (QD) Langmuir monolayer. The deposition is performed via Langmuir–Blodgett (LB) technique. For the sensor chip preparation, covalent immobilization of the thiol-terminated DNA probe sequence (pDNA) using displacement reaction is accomplished. This integrated SPR chip has been used to detect target complementary DNA concentration by monitoring the change in coupling angle via hybridization. It is revealed that this biosensor exhibits high sensitivity (0.7859m0pM−1) towards complementary DNA and can be used to detect it in the concentration range, 180pM to 5pM with detection limit as 4.21pM. The results of kinetic studies yield the values of hybridization and dissociation rate constants as 9.6×104M−1s−1 and 2.3×10−2s−1, respectively, with the equilibrium constant for hybridization as 4.2×106M−1.

Graphene–MoS2 hybrid nanostructures enhanced surface plasmon resonance biosensors

In this work, we propose a new configuration of surface plasmon resonance (SPR) sensor that is based on graphene–MoS2 hybrid structures for ultrasensitive detection of molecules. The proposed system displays a phase-sensitivity enhancement factor of more than 500-fold when compared to the SPR sensing scheme without the graphene–MoS2 coating or with only graphene coating. Our hypothesis is that the monolayer MoS2 has a much higher optical absorption efficiency (∼5%) than that of the graphene layer (∼2.3%). Based on our findings, the electron energy loss of MoS2 layer is comparable to that of graphene and this will allow a successful (∼100%) of light energy transfer to the graphene–MoS2 coated sensing substrate. Such process will lead to a significant enhancement of SPR signals. Our simulation shows that a quasi-dark point of the reflected light can be achieved under this condition and this has resulted in a steep phase jump at the resonance angle of our newly proposed SPR system. More importantly, we found that phase interrogation detection approach of the graphene–MoS2 hybrid structures-based sensing system is more sensitive than that of using the regularly angular interrogation method and our theoretical analysis indicates that 45nm of Au film thickness and 3 coating layers of MoS2 nanosheet are the optimized parameters needed for the proposed SPR system to achieve the highest detection sensitivity range.

Aptamer/thrombin/aptamer-AuNPs sandwich enhanced surface plasmon resonance sensor for the detection of subnanomolar thrombin

A sensitive and selective aptamer/thrombin/aptamer-AuNPs sandwich enhanced surface plasmon resonance (SPR) sensor has been developed for real-time detection of subnanomolar thrombin. In this protocol, one thiol-modified thrombin aptamer (TBA29) was immobilized on gold nanoparticles (AuNPs) via Au–S bonding. The other biotinylated thrombin aptamer (TBA15) was grafted onto streptavidin pretreated SPR gold film through biotin–streptavidin recognition. The presence of thrombin would then induce the formation of a double aptamer sandwich structure on the SPR gold film and results in obvious enhancement of SPR signal, which was proportional to the concentration of thrombin. This proposed assay took advantage of sandwich binding of two affinity aptamers for increased specificity, AuNPs for signal enhancement, as well as SPR signal readout for real-time detection. The SPR signal had a good linear relationship with thrombin concentration in the range of 0.1–75nM, and the detection limit for thrombin was determined to be as low as 0.1nM. It was found that aptamer functionalized AuNPs enhanced the signal of SPR response and thus increased the limit of detection 4-fold and 5-fold compared to direct detection format without AuNPs. This sensor also showed good selectivity for thrombin without being affected by some other proteins, such as BSA and lysozyme. Furthermore, this proposed SPR sensing platform was successfully applied to thrombin analysis in diluted human serum samples.

Detection of Membrane-Binding Proteins by Surface Plasmon Resonance with an All-Aqueous Amplification Scheme

We report here a surface plasmon resonance (SPR) method for detection of cell membrane binding proteins with high degree signal amplification carried out in an all-aqueous condition. Ultrahigh detection sensitivity was achieved for a membrane-based biosensing interface through the use of functional gold nanoparticles (AuNP) in combination with in situ atom transfer radical polymerization (ATRP) reaction. Fusion of phosphatidylcholine vesicles on a calcinated SPR gold chip established a supported bilayer membrane in which cell receptor monosialoganglioside GM1 was embedded for capture of bacterial cholera toxin (CT). The surface-bound CT was recognized with biotinylated anti-CT, which was linked to the biotin-AuNP through an avidin bridge. The biotin-AuNP surface was functionalized with ATRP initiator that triggers localized growth of poly(hydroxyl-ethyl methacrylate) (PHEMA) brush, contributing to marked SPR signal enhancement and quantitative measurement of CT at very low concentrations. The resulting polymer film has been characterized by optical and atomic force microscopy. A calibration curve for CT detection has been obtained displaying a response range from 6.3 × 10(-16) to 6.3 × 10(-8) M with a detection limit of 160 aM (equivalent to ~9500 molecules in 100 μL sample solution). Sensitive detection of biomolecules in complex medium has been conducted with CT-spiked serum, and the detection limit can be effectively improved by 6 orders of magnitude compared to direct measurement in serum. The combined AuNP/ATRP method reported here opens new avenues for ultrasensitive detection of proteins on delicate sensor interfaces constructed by lipid membranes or cell membrane mimics.

Point mutation detection with the sandwich method employing hydrogel nanospheres by the surface plasmon resonance imaging technique

We propose a surface modification procedure to construct DNA arrays for use in surface plasmon resonance (SPR) imaging studies for the highly sensitive detection of a K- ras point mutation, enhanced with hydrogel nanospheres. A homobifunctional alkane dithiol was adsorbed on Au film to obtain the thiol surface, and ethyleneglycol diglycidylether (EGDE) was reacted to insert the ethyleneglycol moiety, which can suppress nonspecific adsorption during SPR analysis. Then streptavidin (SA) was immobilized on EGDE using tosyl chloride activation. Biotinylated DNA ligands were bound to the SA surface via biotin–SA interaction to fabricate DNA arrays. In SPR analysis, the DNA analyte was exposed on the DNA array and hybridized with the immobilized DNA probes. Subsequently, the hydrogel nanospheres conjugated with DNA probes were bound to the DNA analytes in a sandwich configuration. The DNA-carrying nanospheres led to SPR signal enhancement and enabled us to discriminate a K- ras point mutation in the SPR difference image. The application of DNA-carrying hydrogel nanospheres for SPR imaging assays was a promising technique for high throughput and precise detection of point mutations.