Changes In Surface Plasmon Resonance Signal Research Articles

Surface plasmon resonance biosensor for the detection of miRNAs by combining the advantages of homogeneous reaction and heterogeneous detection

The hybridization and enzymolysis reactions for nucleic acid detection were carried out on the chip surface in the traditional surface plasmon resonance (SPR) biosensors. Herein, we proposed an innovative method for microRNA (miRNA) detection in which the hybridization-enzymolysis recycling reactions were performed in solution. Duplex-specific nuclease (DSN) and streptavidin-modified gold nanoparticles (SA-AuNPs) were employed for enhancing the assay sensitivity. In the absence of miRNA, the biotinylated DNA probe (bio-DNA-bio, biotin tags at both the 3′ and 5′ termini of DNA) was attached to the SA-modified chip through the SA-biotin binding, allowing the capture of SA-AuNPs with the same interaction. As a result, a larger SPR signal was attained. However, in the presence of miRNA, bio-DNA-bio hybridized with miRNA was digested by DSN. In this process, the miRNA strand remained intact and participated in the next hybridization-enzymolysis recycling process. Thus, one miRNA could promote the hydrolysis of many bio-DNA-bio probes and allow the generation of numerous bio-DNA fragments. Meanwhile, the produced bio-DNA competed with the undigested bio-DNA-bio to bind SA on the chip surface. The digestion of bio-DNA-bio and the competitive binding between bio-DNA-bio and bio-DNA led to the attachment of fewer SA-AuNPs and then smaller SPR signals. The change in SPR signal at the concentration as low as 1 fM miRNA has been readily determined. The strategy possessed the advantageous properties of simple operation, fast response, high sensitivity and excellent specificity, serving as a viable means for the fabrication of novel sensing platforms.

Real-Time Evaluation of Live Cancer Cells by an in Situ Surface Plasmon Resonance and Electrochemical Study.

This work presents a new strategy of the combination of surface plasmon resonance (SPR) and electrochemical study for real-time evaluation of live cancer cells treated with daunorubicin (DNR) at the interface of the SPR chip and living cancer cells. The observations demonstrate that the SPR signal changes could be closely related to the morphology and mass changes of adsorbed cancer cells and the variation of the refractive index of the medium solution. The results of light microscopy images and 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide studies also illustrate the release or desorption of HepG2 cancer cells, which were due to their apoptosis after treatment with DNR. It is evident that the extracellular concentration of DNR residue can be readily determined through electrochemical measurements. The decreases in the magnitudes of SPR signals were linearly related to cell survival rates, and the combination of SPR with electrochemical study could be utilized to evaluate the potential therapeutic efficiency of bioactive agents to cells. Thus, this label-free, real-time SPR-electrochemical detection technique has great promise in bioanalysis or monitoring of relevant treatment processes in clinical applications.

Graphene based fiber optic surface plasmon resonance for bio-chemical sensor applications

In this study, a surface plasmon resonance (SPR) based fiber optic sensor coated with graphene is introduced. A graphene film synthesized by thermal chemical vapor deposition (TCVD) is transferred onto the sensing area of the optical fiber. The detection mechanism of this sensor is based on the principle that the SPR signal changes according to the refractive indices of analytes. Biotinylated Double Crossover DNA (DXB) lattice and protein Streptavidin (SA) were used for the evaluation. To the best of our knowledge, this is the first attempt to use graphene as a replacement for conventional metal films. The exact SPR phenomena and the red-shift of 7.276nm for the DXB and SA combination were observed.

Plasmonic Diagnostics for Tribology: In Situ Observations Using Surface Plasmon Resonance in Combination with Surface-Enhanced Raman Spectroscopy

The generation of transfer films is a common wear and lubrication mechanism of solid lubricants, such as polymers and lamellar solids. Material can transfer from a solid lubricant to a counter surface as early as the first cycle of sliding, initiating the formation of a transfer film, which can persist for the duration of sliding. Surface plasmon resonance (SPR) was used to monitor incipient molecular wear of a solid lubricant as performed here using a gold coated (50 nm) quartz prism in situ during sliding experiments. Surface-enhanced Raman spectroscopy (SERS) is a complementary technique enabling the analysis of ultra-thin transfer films. SPR and SERS experiments confirm that polytetrafluoroethylene and graphite transfer readily, with observed changes in SPR signal after one cycle of sliding, while ultra-high-molecular-weight polyethylene shows little transfer to the gold during sliding in the in situ SPR experiment. This shows the feasibility of SPR and SERS as important diagnostic tools for tribological studies.

Investigations of electrochemical polymerization processes of thin poly(pyrrole) films and its application to anion sensor based on surface plasmon resonance

A study of the response behavior of anion to surface plasmon resonance (SPR) based sensor, in which a Poly(pyrrole) (Ppy) modified thin gold film was used as a sensor chip is described. In situ surface plasmon resonance and electrochemical methods were used to investigate the electropolymerization and doping/dedoping processes of thin Ppy film. The electropolymerization of pyrrole was carried out under cyclic voltammetric conditions, and simultaneously monitored by in situ SPR. It has revealed that the transition between the reduced state and oxidized state of the Ppy, corresponding to the doping/dedoping of anions, can lead to very distinct changes in SPR signal at a fixed angle of incident laser beam. Furthermore, it has demonstrated that the concentration, the charge and size of anions, as well as the film thickness play important roles in the ingress/egress process of anions. Based on this, this combination of experimental approaches can be used to detect Cl−. SPR signal exhibited a good linear relationship with the concentration of Cl−.

Detection of oligonucleotide systematic mismatches with a surface plasmon resonance sensor

The detection and parallel characterization of the hybridization event of 21-base, unlabeled DNA oligonucleotides with a monolayer of complementary DNA immobilized on a gold surface by surface plasmon resonance (SPR) is presented. A thiol modification on the probe DNA strand allowed for its attachment to the surface via self-assembly. For the hybridization of full match DNA a detection limit of 20 pM was determined. The change in SPR signal was always larger for the full match compared to the one-mismatch target DNA oligonucleotides when the mismatch was in the middle or at the proximal end of the target DNA. Hybridization conditions (buffer concentration, flow rate, and temperature) did not affect the ability of the sensor to discriminate for single nucleotide mismatches. To our knowledge this is the only work where a comparison on the surface hybridization efficiency is performed between proximal, distal, and middle mismatches and the effect of three hybridization parameters is studied with regard to the detection of single nucleotide mismatches using SPR.

Enhanced surface plasmon resonance by Au nanoparticles immobilized on a dielectric SiO 2 layer on a gold surface

This paper introduces strategies for enhancement of a surface plasmon resonance (SPR) signal by adopting colloidal gold nanoparticles (AuNPs) and a SiO 2 layer on a gold surface. AuNPs on SiO 2 on a gold surface were compared with an unmodified gold surface and a SiO 2 layer on a gold surface with no AuNPs attached. The modified surfaces showed significant changes in SPR signal when biomolecules were attached to the surface as compared with an unmodified gold surface. The detection limit of AuNPs immobilized on a SPR chip was 0.1 ng mL −1 for the prostate-specific antigen (PSA), a cancer marker, as measured with a spectrophotometer. Considering that the conventional ELISA method can detect ∼10 ng mL −1 of PSA, the strategy described here is much more sensitive (∼100 fold). The enhanced shift of the absorption curve resulted from the coupling of the surface and particle plasmons by the SiO 2 layer and the AuNPs on the gold surface.

Nondestructive Monitoring of the Photochromic State of Dithienylethene Monolayers by Surface Plasmon Resonance

The use of surface plasmon resonance (SPR) as a nondestructive, nonerasing readout of the isomerization state of a photochromic dithienylethene covalently linked to a chemically modified gold surface was investigated. Four different binding layers were examined: 11-mercaptoundecanol (MUO), an amine-modified 11-mercaptoundecanol (MUO-NH2), dextran, and an amine-modified dextran. The binding of dithienylethene to the modified gold surface and photoisomerization of the photochrome in the bound state were established by FTIR. Solvent effects were measured for every layer tested using ethanol and hexanes. In general, large, easily measurable SPR signal changes could be detected under conditions where photoisomerization of the dithienylethene photochrome was not quenched by the gold plasmon, establishing SPR as a viable form of readout for potential dithienylethene-based optical data storage or processing devices. Dextran-bound photochrome in ethanol exhibited the largest SPR response upon photoisomerization, but is more prone to time-dependent fluctuations resulting from swelling of the dextran layer (caused by slow diffusion of the solvent) than the other layers. Large responses are also provided by MUO-NH2 and MUO, and the signal is much more stable than that for dextran.

The characterization of biomolecular secondary structures by surface plasmon resonance.

Recently there has been considerable interest in using surface plasmon resonance (SPR) for the measurement of conformational changes of immobilized biomolecules that are induced by an exogenous analyte. While a number of studies have shown the analytical utility of such measurements, there has been no report which characterizes the specific secondary structure that actuates the change in SPR signal. The use of SPR to indicate the type of secondary structure present in two immobilized polypeptides, poly-L-lysine (PL) and poly-L-glutamic acid (PGA), and a globular protein, concanavalin A (Con A) is described in this report. The PL, PGA and Con A were modified with N-succinimidyl 3-(2-pyridyldithiol) propionate (SPDP) to introduce disulfide groups to facilitate the attachment onto gold-coated surfaces via self-assembly. Ethanol and 2,2,2-trifluoroethanol (TFE) were used to induce changes in the secondary structure of the immobilized polypeptides and the protein respectively. Using both circular dichroism (CD) and Fourier transform infrared (FTIR) spectroscopies, it has been demonstrated that it is possible to correlate the signal changes observed in SPR to the secondary conformation of the biomolecule. Both CD and FTIR showed that a decrease in SPR signal corresponded to a high content of beta, turn or unordered structures while an increase corresponded to a high alpha-helical content. The sensitivity of the SPR technique is comparable to that obtained in solution with CD and FTIR spectroscopies. These results are the first demonstration that SPR can be used to characterize secondary structures. There is potential, therefore, for SPR to be used as a technique to study secondary conformational changes of immobilized polypeptides and proteins.

Monitoring of the refolding process for immobilized firefly luciferase with a biosensor based on surface plasmon resonance.

In order to examine the possibility of the use of a surface plasmon resonance (SPR) sensor for real-time monitoring of the process of refolding of immobilized proteins, the refolding of firefly luciferase immobilized on a carboxymethyldextran matrix layer was analyzed. The SPR signal of the immobilized luciferase decreased after unfolding induced by GdnCl and increased gradually in the refolding buffer, while there was no signal change in the reference surface lacking the immobilized protein. The decrease in the SPR signal on unfolding was consistent with the difference between the refractive indices of the native and unfolded protein solutions. The effects of blocking of the excess NHS-groups of the matrix layer on the refolding yield were examined by means of an SPR sensor. The results were consistent with those obtained with the enzymatic activity assay, indicating that the changes in the SPR signal reflected the real-time conformational changes of the immobilized protein. Hence, an SPR biosensor might be used for monitoring of the process of refolding of immobilized proteins and as a novel tool for optimization of the refolding conditions. This is the first demonstration that SPR signal changes reflect the conformational changes of an immobilized protein upon unfolding and refolding.