Sensitive Surface Plasmon Resonance Detection Research Articles

DNA walking system integrated with enzymatic cleavage reaction for sensitive surface plasmon resonance detection of miRNA

Abnormal expression levels of miRNA are associated with various tumor diseases, for example, glioma tumors are characterized by the up-regulation of miRNA-182. Surface plasmon resonance (SPR) assay for miRNA-182 from glioma patients was performed via DNA walking amplification strategy. The duplex between aminated swing arm DNA (swDNA) and block DNA (blDNA), and aminated track DNA (trDNA) with a biotin tag were tethered on the poly(ethylene glycol) (PEG)-modified chips. Upon formation of miRNA/blDNA duplex, the SPR signal decreased with the walking process of swDNA, as the biotinylated fragment of trDNA (biotin-TTGGAGT) was detached from the sensor surface caused by the nicking endonuclease Nb.BbvCI. Such a repeated hybridization and cleavage cycle occurred continuously and the detachment of more biotinylated fragments of trDNA from the chips led to the attachment of fewer streptavidin (SA) molecules and then smaller SPR signals. MiRNA-182 with concentrations ranging from 5.0 fM to 1.0 pM could be readily determined and a detection limit of 0.62 fM was achieved. The proposed method was highly selective and possessed remarkable capability for evaluating the expression levels of miRNA-182 in serum samples from healthy donors and glioma patients. The sensing protocol holds great promise for early diagnosis of cancer patients.

A new ultralow fouling surface for the analysis of human plasma samples with surface plasmon resonance

Surface plasmon resonance (SPR) has been widely used to detect a variety of biomolecular systems, but only a small fraction of applications report on the analysis of patients’ samples. A critical barrier to the full implementation of SPR technology in molecular diagnostics currently exists for its potential application to analyze blood plasma or serum samples. Such capability is mostly hindered by the non-specific adsorption of interfering species present in the biological sample at the functional interface of the biosensor, often referred to as fouling. Suitable polymeric layers having a thickness ranging from 15 and about 70 nm are usually deposited on the active surface of biosensors to introduce antifouling properties. A similar approach is not fully adequate for SPR detection where the exponential decay of the evanescent plasmonic field limits the thickness of the layer beyond the SPR metallic sensor surface for which a sensitive detection can be obtained. Here, a triethylene glycol (PEG(3))-pentrimer carboxybetaine system is proposed to fabricate a new surface coating bearing excellent antifouling properties with a thickness of less than 2 nm, thus compatible with sensitive SPR detection. The high variability of experimental conditions described in the literature for the quantitative assessment of the antifouling performances of surface layers moved us to compare the superior antifouling capacity of the new pentrimeric system with that of 4-aminophenylphosphorylcholine, PEG-carboxybetaine and sulfobetaine-modified surface layers, respectively, using undiluted and diluted pooled human plasma samples. The use of the new coating for the immunologic SPRI biosensing of human arginase 1 in plasma is also presented.

Improving nanoparticle-enhanced surface plasmon resonance detection of small molecules by reducing steric hindrance via molecular linkers

Ultrasensitive Surface Plasmon Resonance (SPR) detection of small molecules can be achieved using nanoparticles to both enhance the SPR signals and pre-concentrate low levels of target analytes in the sample. However, the short effective penetration depth of the SPR evanescent field, and steric hindrance of binding when immobilizing small molecules on surfaces, limits the applicability of using relatively large nanoparticles (≥100 nm) for SPR detection. To overcome the issues of steric hindrance, this paper investigates the role of the molecular linkers to tether both the antibodies to the magnetic nanoparticles, and to bind the small molecules to the surface of the SPR chips. By extending the distance of the small molecule (progesterone) away from the SPR chip surface and improving the antibody orientation on the large magnetic nanoparticles, a sensitive SPR detection for progesterone was achieved in buffer (0.013 ng mL−1). The results of the SPR assay for progesterone in milk were in good correlation with ELISA results, and could be used to verify the onset of estrus in cows.

Sensitive surface plasmon resonance detection of methyltransferase activity and screening of its inhibitors amplified by p53 protein bound to methylation-specific ds-DNA consensus sites

Abnormal DNA methylation is closely related to cancer initiation and progression, and strategies to assay methyltransferase activity and screen its inhibitors are essential for cancer diagnosis and therapy. In this work, surface plasmon resonance (SPR)-based assay for real-time and sensitive monitoring of DNA methyltransferase activity and screening of its inhibitors was conducted via methylation of double-stranded (ds)-DNA consensus sites and the follow-up p53 protein recognition. The consensus ds-DNA possesses a specific sequence of 5′-CCGG-3′ in which the second C base can be methylated by M.SssI methyltransferase (M.SssI MTase) and the methylation process impedes the recognition and cleavage of the ds-DNA by HpaII endonuclease, thus, the attachment of p53 protein leads to remarkable SPR signals. In contrast, inhibition of M.SssI MTase activity by a potent inhibitor leaves the consensus ds-DNA unmethylated, and the cleavage of the ds-DNA by HpaII prevents p53 protein from adsorbing onto the chip surface, leading to tiny SPR signals. The binding affinity (KD) between p53 protein and the methylated consensus ds-DNA was deduced to be 3.04 nM, evidencing the strong binding capability. Two nucleoside inhibitors of 5-Azacytidine (5-Aza) and 5-aza-2′-deoxycytidine (5-Aza-dC), and a non-nucleoside inhibitor of procaine were examined, and their half-maximal inhibiting concentration (IC50) values were highly comparable with those by other methods. The sensing protocol has been successfully utilized for the assay of M.SssI MTase activity in normal and cancer cell lysates. The proof-of-concept experiments demonstrate that SPR serves as a viable means for sensitive detection of methyltransferase activity and screening of its inhibitors using p53 protein bound to methylation-specific ds-DNA consensus sites.

A novel and highly specific phage endolysin cell wall binding domain for detection of Bacillus cereus.

Rapid, specific and sensitive detection of pathogenic bacteria is crucial for public health and safety. Bacillus cereus is harmful as it causes foodborne illness and a number of systemic and local infections. We report a novel phage endolysin cell wall-binding domain (CBD) for B. cereus and the development of a highly specific and sensitive surface plasmon resonance (SPR)-based B. cereus detection method using the CBD. The newly discovered CBD from endolysin of PBC1, a B. cereus-specific bacteriophage, provides high specificity and binding capacity to B. cereus. By using the CBD-modified SPR chips, B. cereus can be detected at the range of 10(5)-10(8)CFU/ml. More importantly, the detection limit can be improved to 10(2)CFU/ml by using a subtractive inhibition assay based on the pre-incubation of B. cereus and CBDs, removal of CBD-bound B. cereus, and SPR detection of the unbound CBDs. The present study suggests that the small and genetically engineered CBDs can be promising biological probes for B. cereus. We anticipate that the CBD-based SPR-sensing methods will be useful for the sensitive, selective, and rapid detection of B. cereus.

On-chip microbial culture for the specific detection of very low levels of bacteria

Microbial culture continues to be the most common protocol for bacterial detection and identification in medicine and agronomics. Using this process may take days to identify a specific pathogen for most bacterial strains. Surface Plasmon Resonance (SPR) detection is an emerging alternative technology that can be used for the detection of bacteria using protein microarrays although typical limits of detection are in the range of 10(3)-10(6) cfu mL(-1), which is not compatible with most Food Safety regulation requirements. In this work, we combine concomitant "on-chip" microbial culture with sensitive SPR detection of bacteria thus allowing rapid specific detection of bacteria pathogens - including Salmonella enterica serovar Enteritidis, Streptococcus pneumoniae and Escherichia coli O157:H7 - cultured on a protein microarray. This Culture-Capture-Measure (CCM) approach significantly decreases both the number of processing steps and the overall assay time for bacterial detection. Signal analysis of SPR responses allowed the fast and quantitative assessment of bacterial concentrations initially present in the sample as low as 2.8 ± 19.6 cfu per milliliter. Altogether, our results show how simple, easy-to-operate, fluidic-less and lo-tec microarrays can be used with unprocessed samples and yield - in a single assay - both qualitative and quantitative information regarding bacterial contamination.

Nanogap-based dielectric-specific colocalization for highly sensitive surface plasmon resonance detection of biotin-streptavidin interactions

We have performed highly sensitive surface plasmon resonance (SPR) detection by colocalizing the evanescent near-fields and target molecular distribution. The colocalization is based on oblique metal evaporation to form nanogaps of a size under 100 nm without using electron-beam lithography. The concept was demonstrated by detecting siloxane-based biotin/streptavidin interactions. 50-nm nanogaps produced the largest amplification of optical signatures and two orders of magnitude enhancement of sensitivity over conventional thin film-based measurements. The enhancement is associated with efficient overlap of localized near-fields and target. Colocalized detection scheme is expected to provide clues to molecular sensitivity for SPR biosensing.

Numerical evaluation of periodic nanowire-based phase sensitive surface plasmon resonance detection

Localized surface plasmon (SP)-based amplification of sensitivity in a phase-sensitive SP resonance (SPR) biosensor is investigated numerically. SP is localized by surface-relief periodic nanowires. An optimum sample obtained by modeling DNA hybridization and water–glycerin mixture employed a gold grating with 300 nm period and 10 nm thickness on a gold film and an SF10 glass substrate. The localized SPR structure was shown to have the sensitivity enhanced by more than 10 times than conventional SPR for detecting DNA hybridization at an identical film thickness of 40 nm. The enhancement was found to be associated with field localization near the nanostructure. The results suggest a novel approach of improving the sensitivity of SPR detection.

Molecularly imprinted nanocomposites for highly sensitive SPR detection of a non-aqueous atrazine sample

A surface plasmon resonance (SPR) sensor highly sensitive and selective for the herbicide atrazine was composed by immobilizing atrazine-imprinted polymer with gold nanoparticles on a gold thin film as a sensor chip. In the detection, the atrazine-imprinted polymer was expected to work as synthetic receptor for selectively capturing atrazine in organic solvent, and the gold nanoparticles were expected to exhibit a coupling effect with the gold thin film to enhance the local electromagnetic field between the nanoparticles and the gold film, making the sensor chip highly sensitive for changes in microenvironmental polarity. Thus, a combination of the atrazine-imprinted polymer and gold nanoparticles enabled us to compose an SPR sensor demonstrating the detection of 5 pM atrazine in acetonitrile.

Electroless-plated gold films for sensitive surface plasmon resonance detection of white spot syndrome virus

The paper describes the rapid and label-free detection of the white spot syndrome virus (WSSV) using a surface plasmon resonance (SPR) device based on gold films prepared by electroless plating. The plating condition for obtaining films suitable for SPR measurements was optimized. Gold nanoparticles adsorbed on glass slides were characterized by transmission electron microscopy (TEM). Detection of the WSSV was performed through the binding between WSSV in solution and the anti-WSSV single chain variable fragment (scFv antibody) preimmobilized onto the sensor surface. Morphologies of the as-prepared gold films, gold films modified with self-assembled alkanethiol monolayers, and films covered with antibody were examined using an atomic force microscope (AFM). To demonstrate the viability of the method for real sample analysis, WSSV of different concentrations present in a shrimp hemolymph matrix was determined upon optimizing the surface density of the antibody molecules. The SPR device based on the electroless-plated gold films is capable of detecting concentration of WSSV as low as 2.5 ng/mL in 2% shrimp hemolymph, which is one to two orders of magnitude lower than the level measurable by enzyme-linked immunosorbant assays.