Surface Plasmon Resonance Technique Research Articles

Ultrasensitive DNA Origami Plasmon Sensor for Accurate Detection in Circulating Tumor DNAs

Ultrasensitive DNA Origami Plasmon Sensor for Accurate Detection in Circulating Tumor DNAs

High spatial resolution surface plasmon resonance imaging using a plasmonic chip.

The surface plasmon resonance (SPR) technique has been widely applied to biosensing technologies for the rapid quantification of biomolecules without enzyme and fluorescent labeling. However, the conventional prism-coupling SPR method generally has a detection area of a few mm2, and the large contribution of the background signal forms a barrier to highly sensitive detection. Based on a highly spatially resolved SPR method, the present study constructed a scanning GC-SPR imaging instrument using an objective lens with a high numerical aperture and a plasmonic chip that could be used for grating-coupled SPR. Focusing light on the diffraction limit can suppress background signals and improve detection sensitivity. SPR imaging can also be performed by scanning a focal spot. Using this method, the refractive index of a mixture of water and dimethyl sulfoxide was measured with a detection accuracy of 2.43 × 10-3 RIU. Polydopamine films prepared with a thickness of <5nm were also measured, and each film thickness was evaluated with high sensitivity from the effective refractive index detected in a small area of <1 µm2.

Study of cell and drug interactions based on dual-mode detection using SPR and fluorescence imaging

The investigation of the interactions between cells and drugs forms a crucial aspect of biological and clinical medical studies. Generally, single-cell or local-cellular studies require a microscopic imaging system with high magnifications, which suffers from low detection throughputs and poor time responses. The study presented in this paper combined SPR and fluorescence to achieve cell localization, real-time monitoring of cell images and quantitative analysis of drugs. In order to obtain more comprehensive, accurate and real-time data, a dual-mode system based on surface plasmon resonance (SPR) and fluorescence was constructed based on a 4× magnification lens. This enables simultaneous studies of an entire cell and a specific region of the cell membrane. An adaptive adjustment algorithm was established for distorted SPR images, achieving temporal and spatial matching of the dual-mode detection. The combination of SPR and fluorescence not only achieved micro-detection but also complemented the qualitative or quantitative limitations of SPR or fluorescence method alone. In system characterization, the response signal of SPR was noticed to increase with the increasing concentration of EGF in stimulated cells. It indicated that this platform could be employed for quantitative detection of the cell membrane region. Upon addition of EGF, a peak in the SPR curve was observed, and the cells in the corresponding SPR image turned whiter. This indicated that the platform can simultaneously monitor the SPR response signal and image changes. The response time of fluorescence in EGF testing was several seconds earlier than SPR, revealing that signal transduction first occurred in the whole cell and then propagated to the cell membrane region. The inhibitory ability of Gefitinib on cells was verified in a fast and real-time manner within 20 min. The results indicated that the detection limit of this method was 20 IU/mL for EGF and 10 µg/mL for Gefitinib. In conclusion, this study demonstrates the advantages of SPR and fluorescence dual-mode techniques in the analysis of cell-drug interactions, as well as their strong potential in drug screening.

Metformin blocks BIK1-mediated CPK28 phosphorylation and enhances plant immunity

IntroductionMetformin (MET), derived from Galega officinalis, stands as the primary first-line medication for the treatment of type 2 diabetes (T2D). Despite its well-documented benefits in mammalian cellular processes, its functions and underlying mechanisms in plants remain unclear. ObjectivesThis study aimed to elucidate MET’s role in inducing plant immunity and investigate the associated mechanisms. MethodsTo investigate the impact of MET on enhancing plant immune responses, we conducted assays measuring defense gene expression, reactive oxygen species (ROS) accumulation, mitogen-activated protein kinase (MAPK) phosphorylation, and pathogen infection. Additionally, surface plasmon resonance (SPR) and microscale thermophoresis (MST) techniques were employed to identify MET targets. Protein-protein interactions were analyzed using a luciferase complementation assay and a co-immunoprecipitation assay. ResultsOur findings revealed that MET boosts plant disease resistance by activating MAPKs, upregulating the expression of downstream defense genes, and fortifying the ROS burst. CALCIUM-DEPENDENT PROTEIN KINASE 28 (CPK28) was identified as a target of MET. It inhibited the interaction between BOTRYTIS-INDUCED KINASE 1 (BIK1) and CPK28, blocking CPK28 threonine 76 (T76) transphosphorylation by BIK1, and alleviating the negative regulation of immune responses by CPK28. Moreover, MET enhanced disease resistance in tomato, pepper, and soybean plants. ConclusionCollectively, our data suggest that MET enhances plant immunity by blocking BIK1-mediated CPK28 phosphorylation.

Screening TLR4 Binding Peptide from Naja atra Venom Glands Based on Phage Display.

Toll-like receptor 4 (TLR4) is a crucial inflammatory signaling pathway that can serve as a potential treatment target for various disorders. A number of inhibitors have been developed for the TLR4 pathway, and although no inhibitors have been approved for clinical use, most have been screened against the TLR4-MD2 conformation. The venom gland is the organ of venomous snakes that secretes substances that are toxic to other animals. The level of gene transcription in venom glands is different from that in other tissues, includes a large number of biologically active ingredients, and is an important natural resource for the development of new drugs. We constructed a T7 phage display library using the cobra (Naja atra) venom gland from the Guangdong Snake Breeding Plant and performed three rounds of screening with TLR4 as the target, randomly selecting monoclonal phage spots for PCR followed by Sanger sequencing. The obtained sequences were subjected to length analysis, molecular docking, solubility prediction, and stability prediction, and a peptide containing 39 amino acids (NA39) was finally screened out. The BLAST results indicated that NA39 was a sequence in RPL19 (Ribosomal Protein L19). After peptide synthesis, the binding ability of NA39 to TLR4 was verified by the surface plasmon resonance (SPR) technique. In this study, a new peptide that can specifically bind TLR4 was successfully screened from the cobra venom gland cDNA library, further demonstrating the effectiveness of phage display technology in the field of drug discovery.

Label-free and early detection of HER2 breast cancer biomarker based on UiO-66-NH2 modified gold chip (Au/UiO-66-NH2) using surface plasmon resonance technique

Breast cancer is one of the leading causes of cancer death for women worldwide. This cancer occurs when the breast cells grow abnormally and spread uncontrolled. Although there are several treatments, including chemotherapy, radiation, and surgery in a clinical setting to cure cancer, these treatments cannot provide a 100 % success rate in eliminating cancer, mainly due to the late diagnosis of the disease. Thus, early detection is the key to provide better breast cancer treatment and a higher chance for survival. Herein, simple and sensitive biosensor by combining the emerging multifunctional materials of metal-organic frameworks (MOFs) with a label-free surface plasmon resonance (SPR) instrument was developed. Two types of stable Zirconium (Zr)-based MOFs, namely Zr benzene dicarboxylic acid (Zr-BDC or UiO-66) and Zr 2-amino-benzene dicarboxylic acid (Zr-BDC-NH2 or UiO-66-NH2) were synthesized using the solvothermal method. X-ray diffractometer (XRD) and fourier transform infra red (FTIR) confirmed the formation of the prepared UiO-66 and UiO-66-NH2. The electron microscopy and nitrogen adsorption desorption isotherm characterization reveal that the MOFs have a cubic crystal structure with a high surface area of 672.4 m2/g for UiO-66 and 286.1 m2/g for UiO-66-NH2. The MOFs are then functionalized on the SPR gold (Au) chip surface to serve as a matrix of Human Epidermal Growth Factor Receptor 2(HER2) antibody receptors. Interestingly, the functionalized SPR biosensors could detect the HER2 protein target with high sensitivity in the ng/mL range and a remarkable detection limit, which is 0.457 ng/mL for UiO-66-NH2. This LOD value is significantly lower than clinical concentrations of HER2 proteins in human blood (∼15 ng/mL), which indicate that the biosensor can offer the capability of early detection test.

Anti-inflammatory effects of quinolinyl analog of resveratrol targeting TLR4 in MCAO/R ischemic stroke rat model

BackgroundAmong adults, stroke is the main causes of mortality and permanent disability. Neuroinflammation is one of the main causes of stoke-mediated neuronal death. Our previous study revealed that (E)-5-(2-(Quinolin-4-yl) vinyl) benzene-1, 3-diol (RV01), a quinolinyl analog of resveratrol, inhibits microglia-induced neuroinflammation and safeguards neurons from inflammatory harm. The preventive role of RV01 in ischemic stroke and its underlying cellular mechanisms and molecular targets remain poorly understood. PurposeTo investigate whether RV01 alleviates ischemia-reperfusion (I/R) injury by inhibiting microglia-mediated neuroinflammation and determine the potential molecular mechanisms and targets by which RV01 inhibits the I/R-mediated microglia activation. MethodsRat middle cerebral artery occlusion and reperfusion (MCAO/R) and BV-2 or primary microglial cells oxygen-glucose deprivation and reperfusion (OGD/R) models were established. The neurological behavior scores, 2, 3, 5-triphenyl tetrazolium chloride staining and immunofluorescence were used to detect the neuroprotective effect of RV01 in the MCAO/R rats. In addition, the mRNA expression levels of IL-6, TNF-α, and IL-1β were detected to reveal the antineuroinflammatory effect of RV01. Moreover, a western blot assay was performed to explore the protein expression changes in NF-κB-mediated neuroinflammation. Finally, we identified TLR4 as an RV01 target through molecular docking, drug sensitivity target stability analysis, cellular thermal shift analysis, and surface plasmon resonance techniques. ResultsRV01 reduced the infarct volume and neurological deficits, increased the rotarod duration, and decreased the number of rightward deflections in the MCAO/R rats. RV01 inhibited the NF-κB signaling pathway in vitro and in vivo, as demonstrated by the reduction in the transcription factor p65-mediated expression of several inflammatory factors including IL-6, TNF-α, and IL-1β. Further studies showed that its protective effect was associated with targeting the TLR4 protein. Notably, the anti-inflammatory effect of RV01 was markedly reinforced by the TLR4 knockdown, but inhibited by the overexpression of TLR4. Results revealed that the conditioned medium derived from the RV01-treated BV-2 cells significantly decreased the OGD/R-mediated neuronal damage. ConclusionOur results are the first to reveal the protective effects of RV01 on cerebral ischemia, depending on its inhibitory effect on the NF-κB pathway by targeting TLR4. RV01 could be a potential protective agent in ischemic stroke treatment.

A Novel Plasmonic Sensor Based on Dual-Channel D-Shaped Photonic Crystal Fiber for Enhanced Sensitivity in Simultaneous Detection of Different Analytes.

A dual-channel D-shaped photonic crystal fiber (PCF) based plasmonic sensor is proposed in this paper for the simultaneous detection of two different analytes using the surface plasmon resonance (SPR) technique. The sensor employs a 50 nm-thick layer of chemically stable gold on both cleaved surfaces of the PCF to induce the SPR effect. This configuration offers superior sensitivity and rapid response, making it highly effective for sensing applications. Numerical investigations are conducted using the finite element method (FEM). After optimizing the structural parameters, the sensor exhibits a maximum wavelength sensitivity of 10000 nm/RIU and an amplitude sensitivity of -216 RIU -1 between the two channels. Additionally, each channel of the sensor exhibits its unique maximal wavelength and amplitude sensitivities for different refractive index (RI) ranges. Both channels demonstrate a maximal wavelength sensitivity of 6000 nm/RIU. In the RI range of 1.31-1.41, Channel 1 (Ch1) and Channel 2 (Ch2) achieved their maximum amplitude sensitivities of -85.39RIU -1 and -304.52 RIU -1 , respectively, with a resolution of 5×10 -5 . This sensor structure is noteworthy for its ability to measure both amplitude and wavelength sensitivity, providing enhanced performance characteristics suitable for various sensing purposes in chemical, biomedical, and industrial fields.

Optimizing drug discovery: Surface plasmon resonance techniques and their multifaceted applications

In recent years, researchers have made significant contributions to the field of Surface Plasmon Resonance sensors and their potential applications. SPR features include real-time monitoring of biomolecular interactions, label-free detection, high sensitivity, and the ability to analyze multiple samples at the same time. These characteristics make SPR biosensors a valuable tool in the pharmaceutical industry for ensuring drug efficacy and safety. Furthermore, SPR sensors have a reputation for providing quantitative and qualitative superior performance in real-time applications such as drug development, environmental monitoring, food safety, medical diagnostics, clinical diagnosis, biological investigations, and biomolecule interactions. This review will go over recent advances in SPR biosensors and how they can be used in drug discovery and development. We will also look at the challenges and opportunities of using SPR biosensors in the pharmaceutical industry.

Visible and angular interrogation of Kretschmann-based SPR using hybrid Au–ZnO optical sensor for hyperuricemia detection

Uric acid is a waste product of the human body where high levels of it or hyperuricemia can lead to gout, kidney disease and other health issues. In this paper, Finite Difference Time Doman (FDTD) simulation method was used to develop a plasmonic optical sensor to detect uric acid with molarity ranging from 0 to 3.0 mM. A hybrid layer of gold-zinc oxide (Au–ZnO) was used in this Kretschmann-based Surface Plasmon Resonance (K-SPR) technique with angular interrogation at 670 nm and 785 nm visible optical wavelengths. The purpose of this study is to observe the ability of the hybrid material as a sensing performance enhancer for differentiating between healthy and unhealthy uric acid levels based on the refractive index values from previous study. Upon exposure to 670 nm wavelength, the average sensitivity of this sensor was found to be 0.028°/mM with a linearity of 98.67 % and Q-factor value of 0.0053 mM−1. While at 785 nm, the average sensitivity is equal to 0.0193°/mM with slightly lower linearity at 94.46 % and Q-factor value of 0.0076 mM−1. The results have proven the ability of hybrid material Au–ZnO as a sensing performance enhancer for detecting uric acid when compared with bare Au and can be further explored in experimental work.

Study on Saccharide-Glucose Receptor Interactions with the Use of Surface Plasmon Resonance.

The aim of this study was to investigate the process of attachment of saccharide particles differing in degree of complexity to cell receptors responsible for transport of glucose across the cell membrane (GLUT proteins). This phenomenon is currently considered when designing modern medicines, e.g., peptide drugs to which glucose residues are attached, enabling drugs to cross the barrier of cell membranes and act inside cells. This study aims to help us understand the process of assimilation of polysaccharide nanoparticles by tumour cells. In this study, the interactions between simple saccharides (glucose and sucrose) and dextran nanoparticles with two species of GLUT proteins (GLUT1 and GLUT4) were measured using the surface plasmon resonance technique. We managed to observe the interactions of glucose and sucrose with both applied proteins. The lowest concentration that resulted in the detection of interaction was 4 mM of glucose on GLUT1. Nanoparticles were measured using the same proteins with a detection limit of 40 mM. These results indicate that polysaccharide nanoparticles interact with GLUT proteins. The measured strengths of interactions differ between proteins; thus, this study can suggest which protein is preferable when considering it as a mean of nanoparticle carrier transport.

Polymeric carbon quantum dots as efficient chlorophyll sensor-analysis based on experimental and computational investigation

This report introduces a microfiber chlorophyll sensor based on polymeric amine functionalized carbon quantum dots (NCQD) using the surface plasmon resonance technique. The silver (Ag) thin film, followed by NCQD-polyvinyl alcohol composite film, was deposited on the tapered optical fiber. The characterization of surface morphologies of NCQD-PVA film using a field emission scanning electron microscope shows that the contact area with the analyte was well covered with Ag and NCQD-PVA. The experimental results show that the fiber optic SPR sensor can detect chlorophyll in the 0.01–2.0 ppm range with a high sensitivity response and a detection limit of 1.90 nm ppm−1 and 0.78 ppm, respectively. This study observed no significant interference among the other ionic species (Fe3+, NH4+, NO3–, NO2–, and PO4-) due to the electrostatic and π-π interactions between NCQD and chlorophyll. Results from density functional theory calculation also confirm that the interaction of the NCQD with chlorophyll existed, and the strongest interaction occurs between nitrogen at pyridine sites of the carbon core and the oxygen group of chlorophyll. The proposed NCQD/PVA optical fiber sensor exhibits good sensing performance and correlates well with the standard method (R2 = 0.9501), suggesting a suitable technology candidate for real-time environmental monitoring applications.

Detection of Various Traditional Chinese Medicinal Metabolites as Angiotensin-Converting Enzyme Inhibitors: Molecular Docking, Activity Testing, and Surface Plasmon Resonance Approaches.

Angiotensin-converting enzyme 1 (ACE1) is a peptide involved in fluid and blood pressure management. It regulates blood pressure by converting angiotensin I to angiotensin II, which has vasoconstrictive effects. Previous studies have shown that certain compounds of natural origin can inhibit the activity of angiotensin-converting enzymes and exert blood pressure-regulating effects. Surface Plasmon Resonance (SPR) biosensor technology is the industry standard method for observing biomolecule interactions. In our study, we used molecular simulation methods to investigate the docking energies of various herbal metabolites with ACE1 proteins, tested the real-time binding affinities between various herbal metabolites and sACE1 by SPR, and analyzed the relationship between real-time binding affinity and docking energy. In addition, to further explore the connection between inhibitor activity and real-time binding affinity, several herbal metabolites' in vitro inhibitory activities were tested using an ACE1 activity test kit. The molecular docking simulation technique's results and the real-time affinity tested by the SPR technique were found to be negatively correlated, and the virtual docking technique still has some drawbacks as a tool for forecasting proteins' affinities to the metabolites of Chinese herbal metabolites. There may be a positive correlation between the enzyme inhibitory activity and the real-time affinity detected by the SPR technique, and the results from the SPR technique may provide convincing evidence to prove the interaction between herbal metabolites and ACE1 target proteins.

Spectroscopy evidence of interaction of 1,5 disubstituted piperidino-amido anthraquinone derivative with human telomeric G-quadruplex DNA: Basis of anticancer action

The binding of ligands to G-quadruplex (G4) structures at the ends of human telomeric DNA induces thermal stabilization, which interferes with telomere maintenance by disrupting the association of telomeres with the telomerase enzyme—an important marker for cancer. Understanding the binding mode of the ligand-G quadruplex DNA complex is imperative for evaluating the relative efficacy and specificity of their interaction. We focused on the interaction of 1,5-Bis(3-piperidino propionamido) anthracene-9,10-dione with human telomeric DNA sequences using surface plasmon resonance, absorbance, fluorescence (steady-state and lifetime), and circular dichroism spectroscopy techniques. The ligand binding with HTel22 (Kb = 8.4 × 105 M−1) induced cell toxicity with an IC50 value of ∼ 8.64 µM in MCF-7 cancer cell lines. Significant hypochromism (59 %), fluorescence quenching (97 %), no change in fluorescence lifetime and absence of induced Circular Dichroism (CD) band upon addition of G4 DNA to ligand, suggest a groove/external binding mode. CD spectral changes reflect rearrangement in parallel and antiparallel strands to accommodate the ligand. Docking results reveal specific short contacts of the ligand's side chain, including 9CO, 13N, 14NH, and 11CO, with the grooves of the G4 DNA, without making any contact with the loops, favoring an energetically more favorable conformation. Thermal denaturation profiles, obtained by Differential Scanning Calorimetry and CD, show the stabilization of wHTel26 and HTel22 G4 DNA in K+ and Na+ rich solutions by 21.2 and 17.6 °C, respectively, which may restrict the access of telomerase to telomeres. The findings indicate the potential of modifying anthraquinone substituent groups for therapeutic applications.

Graphene - Coated Surface Plasmon Resonance(SPR) Sensor for Detection of Preservatives in Milk : A Theoretical Investigation

Milk is an essential dietary staple for human sustenance. However, the presence of various contaminants, notably preservatives, within milk can potentially pose significant health risks and impact overall well-being. Consequently, the meticulous surveillance of pollutants and impurities, particularly preservatives, in natural and everyday food products has emerged as a pivotal endeavor to ensure food safety and enhance its overall quality. In this research study, the Surface Plasmon Resonance (SPR) technique is strategically employed for a theoretical exploration of preservatives, namely hydrogen peroxide, formaldehyde, and sodium carbonate, within milk. The investigation encompasses varying concentrations of these preservatives, ranging from 0% to 14.3%. Concurrently, the refractive indices span from 1.34550 to 1.35093, representing the concentration between the minimum (0%) and maximum (14.286%) concentrations of hydrogen peroxide, formaldehyde, and sodium carbonate. To authenticate the efficacy of the proposed sensor, an extensive assessment of performance parameters is conducted. These parameters encompass Sensitivity, Full Width at Half Maximum (FWHM), Figure of Merit (FOM), and Detection Accuracy (DA), with calculations performed for each specific case.

A Novel Approach to Realize Plasmonic Sensors via Multimode Optical Waveguides: A Review

In recent decades, the Surface Plasmon Resonance (SPR) phenomenon has been utilized as an underlying technique in a broad range of application fields. Herein, a new measuring strategy which harnesses the SPR technique in a way that is different from the classical methodology was explored by taking advantage of the characteristics of multimode waveguides, such as plastic optical fibers (POFs) or hetero-core fibers. The sensor systems based on this innovative sensing approach were designed, fabricated, and investigated to assess their ability to measure various physical features, such as magnetic field, temperature, force, and volume, and to realize chemical sensors. In more detail, a sensitive patch of fiber was used in series with a multimodal waveguide where the SPR took place, to alter the mode profile of the light at the input of the waveguide itself. In fact, when the changes of the physical feature of interest acted on the sensitive patch, a variation of the incident angles of the light launched in the multimodal waveguide occurred, and, as a consequence, a shift in resonance wavelength took place. The proposed approach permitted the separation of the measurand interaction zone and the SPR zone. This meant that the SPR zone could be realized only with a buffer layer and a metallic film, thus optimizing the total thickness of the layers for the best sensitivity, regardless of the measurand type. The proposed review aims to summarize the capabilities of this innovative sensing approach to realize several types of sensors for different application fields, showing the high performances obtained by exploiting a simple production process and an easy experimental setup.

Mechanistic and kinetic aspects of Natamycin interaction with serum albumin using spectroscopic and molecular docking methods

Natamycin (NT) is a polyene natural antimycotic, which has an antimicrobial effect against yeasts and molds and is used as a preservative in the food industry. In the present study, we evaluated NT interaction with bovine serum albumin (BSA) through surface plasmon resonance (SPR) and several spectroscopic techniques, which are accompanied by a molecular docking study. According to the results, the intensity of BSA fluorescence decreased by adding different concentrations of NT. The fluorescence quenching results showed that NT reduces the intensity of BSA fluorescence by forming a complex with BSA through a hybrid quenching. Binding constant decreases from 18.73 to 2.13 (102 M−1) with increasing temperature, which indicates a decrease in complex formation owing to the interaction of NT with BSA. Negative values of ΔH° and ΔS° confirmed that van der Waals forces and hydrogen bonds are the basic forces in the interaction of NT with BSA. Moreover, increasing the equilibrium constants values with increasing temperature indicated that BSA binding to NT decreased. Finally, BSA interaction occurring with NT through Ser 109, Asp 111, Lys 114, Leu 115, Glu 424, and Arg 458 have been verified via molecular docking analysis. Attained results via SPR and fluorimetry showed that the binding constant between BSA and NT decreased when the temperature was raised.

Molecularly Imprinted Plasmonic Sensors for the Determination of Environmental Water Contaminants: A Review

The scarcity of clean water leads to the exploration of the possibility of using treated wastewater. However, monitoring campaigns have proven the presence of emerging contaminants, such as pharmaceuticals, pesticides and personal care products, not only in trace amounts. Various analytical methodologies have been developed over the last years for the quantification of these compounds in environmental waters. Facing the need to achieve a higher sensitivity, fast response and practical use via miniaturization, the potential of plasmonic sensors has been explored. Through the introduction of molecularly imprinted polymers (MIPs) as recognition elements, MIP-based plasmonic sensors seem to be a good alternative for monitoring a wide range of analytes in water samples. This work attempts to provide a general overview of this form of sensor, which has been reported as being able to sense different contaminants in waters using surface plasmon resonance (SPR) and surface-enhanced Raman-scattering (SERS) techniques. Particular emphasis is given to the fabrication/recognition procedure, including the preparation of MIPs and the use of metals and nanomaterials to increase the performance characteristics of the sensors.

A Straightforward Method for the Development of Positively Charged Gold Nanoparticle-Based Vectors for Effective siRNA Delivery.

The therapeutic potential of short interfering RNA (siRNA) to treat many diseases that are incurable with traditional preparations is limited by the extensive metabolism of serum nucleases, low permeability through biological membrane barriers because of a negative charge, and endosomal trapping. Effective delivery vectors are required to overcome these challenges without causing unwanted side effects. Here, we present a relatively simple synthetic protocol to obtain positively charged gold nanoparticles (AuNPs) with narrow size distribution and the surface modified with Tat-related cell-penetrating peptide. The AuNPs were characterized using TEM and the localized surface plasmon resonance technique. The synthesized AuNPs showed low toxicity in experiments in vitro and were able to effectively form complexes with double-stranded siRNA. The obtained delivery vehicles were used for intracellular delivery of siRNA in an ARPE-19 cell line transfected with secreted embryonic alkaline phosphatase (SEAP). The delivered oligonucleotide remained intact and caused a significant knockdown effect on SEAP cell production. The developed material could be useful for delivery of negatively charged macromolecules, such as antisense oligonucleotides and various RNAs, particularly for retinal pigment epithelial cell drug delivery.

Development of a surface plasmon resonance (SPR) assay for rapid detection of Aeromonas hydrophila

Aquaculture plays an increasingly important if not critical role in the current and future world food supply. Aeromonas hydrophila, a heterotrophic, Gram-negative, bacterium found in fresh or brackish water in warm climates poses a serious threat to the aquaculture industry in many areas, causing significant economic losses. Rapid, portable detection methods of A. hydrophila are needed for its effective control and mitigation. We have developed a surface plasmon resonance (SPR) technique to detect PCR (polymerase chain reaction) products that can replace agarose gel electrophoresis, or otherwise provide an alternative to costlier and more complicated real-time, fluorescence-based detection. The SPR method provides sensitivity comparable to gel electrophoresis, while reducing labor, cross-contamination, and test time, and employs simpler instrumentation with lower cost than real-time PCR.