Biosensor Surface Research Articles

Surface-Functionalized Terahertz Metamaterial Biosensor Used for the Detection of Exosomes in Patients.

Owing to their stability in bodily fluids, exosomes have attracted increased attention as colorectal cancer (CRC) biomarkers for early diagnosis. To validate the potential of the plasma exosomes as a novel biomarker for the monitoring of CRC, we demonstrated a terahertz (THz) metamaterials (MMs) biosensor for the detection of exosomes in this work. The biosensor with two resonant frequencies is designed using full wave electromagnetic simulation software based on the finite integration time domain (FITD) method and fabricated by a surface micromachining process. The biosensor surface is first modified using Au nanoparticles (AuNPs), and then, anti-KRAS and anti-CD147, which are specific to the exosomes, are modified on the AuNPs assembled with HS-poly(ethylene glycol)-COOH (HS-PEG-COOH). Exosomes used in the experiment are extracted via the instructions in the exosomes isolation and purification kit and identified by using transmission electron microscopy (TEM), Western blot (WB), and nanoparticle tracking analysis (NTA). The biosensor covered with plasma-derived exosomes of CRC patients has a different resonance frequency shift compared to that with healthy-control-derived exosomes. This study proposes an emerging and quick method for diagnosing the CRC.

An Effective Label-Free Electrochemical Aptasensor Based on Gold Nanoparticles for Gluten Detection.

Nanomaterials can be used to modify electrodes and improve the conductivity and the performance of electrochemical sensors. Among various nanomaterials, gold-based nanostructures have been used as an anchoring platform for the functionalization of biosensor surfaces. One of the main advantages of using gold for the modification of electrodes is its great affinity for thiol-containing molecules, such as proteins, forming a strong Au-S bond. In this work, we present an impedimetric biosensor based on gold nanoparticles and a truncated aptamer for the quantification of gluten in hydrolyzed matrices such as beer and soy sauce. A good relationship between the Rct values and PWG-Gliadin concentration was found in the range between 0.1–1 mg L−1 of gliadin (corresponding to 0.2–2 mg L−1 of gluten) with a limit of detection of 0.05 mg L−1 of gliadin (corresponding to 0.1 mg L−1 of gluten). The label-free assay was also successfully applied for the determination of real food samples.

Engineering of multifunctional carbon nanodots-decorated plasmonic Au@Ag nanoenzymes for photoelectrochemical biosensing of microRNA-155

Herein, the multifunctional carbon nanodots (CDs)-decorated plasmonic Au@Ag nanoenzymes coupled with Ti2C/Ag2S composites were proposed for photoelectrochemical (PEC) biosensing of microRNA. Two-dimensional (2D) Ti2C MXenes-sensitized Ag2S quantum dots (QDs) (Ti2C/Ag2S composites) acquired an initial photocurrent. After immobilization of a hairpin subsidiary probe (S1) onto the Ti2C/Ag2S composites photoelectrode surface, the target microRNA-associated bipedal DNA walker performed the opening of S1 for forming triple helix molecules. Then the CDs-decorated Au@Ag core–shell (Au@Ag@CDs) nanocuboids were introduced into the PEC biosensing platform through target-induced triple helix molecules. The Au@Ag@CDs nanocuboids as a multifunctional signal amplifier not only competitively captured the irradiating light and electron donors to weaken the PEC signal, but also quenched the photocurrent response resulting from energy transfer between plasmonic bimetallic Au@Ag nanocuboids and Ag2S QDs. Additionally, the Au@Ag and CDs in Au@Ag@CDs nanocuboids displayed a synergistic catalytic effect, and could serve as peroxidase nanoenzymes to catalyze the production of precipitates onto the biosensor surface, resulting in a significantly decreased photocurrent. Based on the plasmonic Au@Ag@CDs nanoenzymes coupled with Ti2C/Ag2S composites, microRNA-155 was ultrasensitively detected with a wide linear range (1–10,000 fM) and a low limit of detection (0.83 fM). Moreover, the engineered plasmonic Au@Ag@CDs nanoenzymes offer an environmentally friendly and multifunctional signal amplifier for fabricating PEC biosensor and have potential applications in biomarkers detection and disease diagnostics.

Influence of enzyme immobilization and skin-sensor interface on non-invasive glucose determination from interstitial fluid obtained by magnetohydrodynamic extraction

We integrated a magnetohydrodynamic fluid extractor with an amperometric glucose biosensor to develop a wearable device for non-invasive glucose monitoring. Reproducible fluid extraction through the skin and efficient transport of the extracted fluid to the biosensor surface are prerequisites for non-invasive glucose monitoring. We optimized the enzyme immobilization and the interface layer between the sensing device and the skin. The monitoring device was evaluated by extracting fluid through porcine skin followed by glucose detection at the biosensor. The biosensor featured a screen-printed layer of Prussian Blue that was coated with a layer containing glucose oxidase. Both physical entrapment of glucose oxidase in chitosan and tethering of glucose oxidase to electrospun nanofibers were evaluated. Binding of glucose oxidase to nanofibers under mild conditions provided a stable biosensor with analytical performance suitable for accurate detection of micromolar concentrations of glucose. Hydrogels of varying thickness (95–2000 μm) as well as a thin (30 μm) nanofibrous polycaprolactone mat were studied as an interface layer between the biosensor and the skin. The effect of mass transfer phenomena at the biosensor-skin interface on the analytical performance of the biosensor was evaluated. The sensing device detected glucose extracted through porcine skin with an apparent (overall) sensitivity of −0.8 mA/(M·cm2), compared to a sensitivity of −17 mA/(M·cm2) for measurement in solution. The amperometric response of the biosensor correlated with the glucose concentration in the fluid that had been extracted through porcine skin with the magnetohydrodynamic technique.

An Electrochemical and Raman Scattering Dual Detection Biosensor for Rapid Screening and Biomolecular Profiling of Cancer Biomarkers

Detecting circulating biomarkers sensitively and quantitatively is paramount for cancer screening, diagnosis, and treatment selection. Particularly, screening of a panel of circulating protein biomarkers followed by mapping of individual biomarkers could assist better diagnosis and understanding of the cancer progression mechanisms. Herein, we present a miniaturized biosensing platform with dual readout schemes (electrochemical and Surface enhanced Raman scattering (SERS)) for rapid cancer screening and specific biomarker expressional profiling to support cancer management. Our approach utilizes a controlled nanomixing phenomena under alternative current electrohydrodynamic condition to improve the isolation of cancer-associated circulating proteins (i.e., Epidermal growth factor receptor (EGFR), BRAF, Programmed death-ligand 1 (PD-L1)) with antibody functionalized sensor surface for rapid and efficient isolation of the targets and subsequent labelling with SERS nanotags. The method employs Differential Pulse Voltammetry (DPV) for rapidly screening for the presence of the circulating proteins on biosensor surface irrespective of their type. Upon positive DPV detection, SERS is applied for sensitive read-out of individual biomarkers biomarker levels. In a proof-of-concept study, we demonstrate the dual detection biosensor for analysing circulating BRAF, EGFR and PDL-1 proteins and successfully screened both ensemble and individual biomarker expressional levels as low as 10 pg (1 ng/mL). Our findings clearly indicate the potential of the proposed method for cancer biomarker analysis which may drive the translation of this dual sensing concept in clinical settings.

Application of silver solid amalgam electrodes in electrochemical detection of DNA damage.

In this study, a mercury meniscus-modified silver solid amalgam electrode was used for the first time for the detection of UV-induced DNA damage. The integrity of the double-stranded DNA (dsDNA) layer was detected indirectly using the evaluation of the methylene blue reduction within its accumulation into dsDNA after the UV irradiation of the biosensor surface with two different wavelengths (254nm and 365nm), monitored by differential pulse voltammetry. Moreover, a simple electrochemical characterization of the biosensor surface was performed using cyclic voltammetry of the redox indicator hexaammineruthenium chloride (RuHex) present in the solution. Electrochemical impedance spectroscopy (EIS) was used in both cases for the verification of results. Individual electrochemical signals depend on the time of biosensor exposure to UV irradiation as well as on the selected wavelengths and are different for both used types of dsDNA (salmon sperm and calf thymus). The highest degradation degree up to 60% was observed using sensitive EIS of methylene blue after 10min irradiation of the biosensor at 254nm. The use of RuHex seems to be less sensitive for the detection of dsDNA structural changes, when the degradation degree up to 40% was observed, using EIS at the same conditions.

Equivalent circuit model of a non-faradaic impedimetric ZnO nano-crystal biosensor

• Sensing of C-reactive protein using a novel co-planar electrode PET-ZnO biosensor. • Development of an EEC model of the biosensor platform. • EEC modelling demonstrates predicted properties of a non-faradaic biosensor. • Quantified EDL charge modulations arising due to CRP binding at the ZnO surface. • Dominance of each sensor component identified to support control of parasitics. This paper describes the development of an equivalent electrical circuit (EEC) model of a unique co-planar electrode PET-ZnO biosensor platform. This non-faradaic ZnO biosensor had a PET insulating layer between the electrodes and the active ZnO sensor surface and impedance spectroscopy was used to measure varying concentrations of C-reactive protein (CRP). Analysis of the impedance magnitude bode plot reveals a slope indicative of a capacitive behaviour, where R 2 is 0.99. Furthermore, extraction of the complex impedance parameters shows capacitive reactance (Xc) being the dominant parameter as a result of capacitive transduction mechanism. The EEC was based on the generalized Randles model and adapted to represent the non-faradaic ZnO biosensor. Experimental data was fitted to the model at 100 kHz to establish values for the component parts of the biosensor system. With these values the model showed good correlation between experimental and modelled data across the frequency range studied, within the limitations of the measurement instrument. The model quantified charge modulations arising due to bimolecular binding of CRP within the electrical double layer (EDL) formed at the ZnO-PBS interface. The CRP bio-molecular interaction within the EDL can cause high charge carrier density accumulation in the semiconductor and as a result the capacitance of the EDL is increased. The EEC model quantifies this phenomenon, presented as a parallel capacitance connected to the ZnO-Antibody interfacial capacitance. The model was adapted to two variants of the biosensor design, with differing ZnO nanoparticle concentrations forming the sensor surface. The variations on the components modelled concurred with expected changes established from biosensor surface analysis. Using the EEC model, the dominance of each constituent sensor element on measurement stability is quantified identifying the significant contributors to experimental and sensor-to-sensor repeatability to support control of parasitic effects.

Mapping the DNA Damaging Effects of Polypyridyl Copper Complexes with DNA Electrochemical Biosensors.

Several classes of copper complexes are known to induce oxidative DNA damage that mediates cell death. These compounds are potentially useful anticancer agents and detailed investigation can reveal the mode of DNA interaction, binding strength, and type of oxidative lesion formed. We recently reported the development of a DNA electrochemical biosensor employed to quantify the DNA cleavage activity of the well-studied [Cu(phen)2]2+ chemical nuclease. However, to validate the broader compatibility of this sensor for use with more diverse—and biologically compatible—copper complexes, and to probe its use from a drug discovery perspective, analysis involving new compound libraries is required. Here, we report on the DNA binding and quantitative cleavage activity of the [Cu(TPMA)(N,N)]2+ class (where TPMA = tris-2-pyridylmethylamine) using a DNA electrochemical biosensor. TPMA is a tripodal copper caging ligand, while N,N represents a bidentate planar phenanthrene ligand capable of enhancing DNA interactions through intercalation. All complexes exhibited electroactivity and interact with DNA through partial (or semi-) intercalation but predominantly through electrostatic attraction. Although TPMA provides excellent solution stability, the bulky ligand enforces a non-planar geometry on the complex, which sterically impedes full interaction. [Cu(TPMA)(phen)]2+ and [Cu(TPMA)(DPQ)]2+ cleaved 39% and 48% of the DNA strands from the biosensor surface, respectively, while complexes [Cu(TPMA)(bipy)]2+ and [Cu(TPMA)(PD)]2+ exhibit comparatively moderate nuclease efficacy (ca. 26%). Comparing the nuclease activities of [Cu(TPMA)(phen)] 2+ and [Cu(phen)2]2+ (ca. 23%) confirms the presence of TPMA significantly enhances chemical nuclease activity. Therefore, the use of this DNA electrochemical biosensor is compatible with copper(II) polypyridyl complexes and reveals TPMA complexes as a promising class of DNA damaging agent with tuneable activity due to coordinated ancillary phenanthrene ligands.

ANTI-miRNA IMMOBILIZATION OPTIMIZATION ON THE SCREEN PRINTED ELECTRODES FOR ELECTROCHEMICAL miRNA BIOSENSORS

Synthetically produced miRNA molecules plays an important role as biomarker to examine and investigate the diagnosis of some diseases including cancer. In order to develop a sensitive electrochemical biosensor system for the detection of miRNA molecules, the anti-miRNA molecules are synthesized and immobilized on the biosensor surfaces and observe the signal changes via a proper measurement. Immobilization time and temperature along with the anti-miRNA concentration are critically important for an appropriate observation of the miRNA detection sensitivity of the prepared biosensor system. In this regard, synthetically produced anti-miRNA (anti-miR451(G)) was purchased and diluted into different concentration by using phosphate buffer solution. Then, the solutions were immobilized on the screen printed electrodes (SPEs) and the guanine oxidation signal of the anti-miRNA molecules were observed via differential pulse voltammetry method (DPV). An appropriate concentration of the solution was selected and dropped on the SPEs and held on at different temperatures (-18, +5 and +25 oC) for 1, 3, 14 and 21 days and DPV measurements were conducted to investigate the optimum immobilization time and temperature. The result shown that guanine oxidation signal was increased by increasing the concentration of the genetic molecules in the immobilization solution and increased less after that point when the concentration increased more because the surface reached to a certain saturation value . The guanine oxidation signal revealed that the best suitable storing temperature after the immobilization was +5 oC determined.

Accelerated Digital Biodetection Using Magneto-plasmonic Nanoparticle-Coupled Photonic Resonator Absorption Microscopy.

Rapid, ultrasensitive, and selective quantification of circulating microRNA (miRNA) biomarkers in body fluids is increasingly deployed in early cancer diagnosis, prognosis, and therapy monitoring. While nanoparticle tags enable detection of nucleic acid or protein biomarkers with digital resolution and subfemtomolar detection limits without enzymatic amplification, the response time of these assays is typically dominated by diffusion-limited transport of the analytes or nanotags to the biosensor surface. Here, we present a magnetic activate capture and digital counting (mAC+DC) approach that utilizes magneto-plasmonic nanoparticles (MPNPs) to accelerate single-molecule sensing, demonstrated by miRNA detection via toehold-mediated strand displacement. Spiky Fe3O4@Au MPNPs with immobilized target-specific probes are "activated" by binding with miRNA targets, followed by magnetically driven transport through the bulk fluid toward nanoparticle capture probes on a photonic crystal (PC). By spectrally matching the localized surface plasmon resonance of the MPNPs to the PC-guided resonance, each captured MPNP locally quenches the PC reflection efficiency, thus enabling captured MPNPs to be individually visualized with high contrast for counting. We demonstrate quantification of the miR-375 cancer biomarker directly from unprocessed human serum with a 1 min response time, a detection limit of 61.9 aM, a broad dynamic range (100 aM to 10 pM), and a single-base mismatch selectivity. The approach is well-suited for minimally invasive biomarker quantification, enabling potential applications in point-of-care testing with short sample-to-answer time.

Multifunctional amino acids empowering bifunctional biosensing platform for depression study

Poor availability of objective approaches hinders effective diagnosis and treatment for depression. Biosensors provide a promising platform for the development of quantitative and practical methods for disease detection, as well as for drug discovery. Here, we developed an electrochemical biosensor has been established with the ability to simply and accurately detect the trace glucocorticoid receptor alpha (GRα), as a key biomarker of depression, in both hippocampus and blood cells. The integration of amino-ion graphene oxide (IL-rGO) and amino acid-coated gold nanoparticles (AA-AuNPs) via green synthesis remarkably magnifies the electrochemical signals, where amino acids play multiple roles as reducing agents, stabilizers, and bridging agents. After the optimization among AA-AuNPs@IL-rGO nanocomposites based on five typical amino acids, a biosensing surface has been constructed to implement analysis in real samples as a bifunctional platform. The obtained biosensor exhibited a remarkably low limit of detection (0.283 pg mL−1) and could thus sensitively identify the GRα differences in healthy and depressive rats with and without fluoxetine. The electrochemical biosensor developed herein was not only outstandingly sensitive but also simple to use and labor-saving, making it a promising all-in-one platform for depression diagnosis and drug discovery.

Front Cover: Enhancing affinity‐based electroanalytical biosensors by integrated AC electrokinetic enrichment—A mini review

DOI: 10.1002/elps.202100168 The cover picture shows how affinity-based electroanalytical biosensors can be enhanced by integrated AC electrokinetic (ACEK) effects. ACEK effects, including AC electrothermal (ACET) effect, AC electroosmosis (ACEO) effect and dielectrophoretic (DEP) effect, are produced by applying AC signals to the interdigitated electrode arrays. ACEK effects induce directed movement of microflows and particles (e.g., proteins, nucleic acids, bacteria, small molecules, etc.) in the solution, so that the analytes in the solution can be quickly routed toward the surface of biosensors, thus improving the binding efficiency between the analytes and the probes on the surface of the sensor. Studies have shown that the method holds great promise for achieving high sensitivity, low detection limit and rapid turnaround.

A bio-sensing surface with high biocompatibility for enhancing Raman scattering signals as enabled by a Mo-Ag film.

Biological surface-enhanced Raman scattering (SERS) sensing is dependent on the properties of the sensing surface. For the biological sensing surfaces for SERS, the hydrophilicity, biocompatibility and signal sensitivity are pivotal. Hence, a hard bimetallic Mo-Ag film was developed by a magnetron sputtering technique, and the surface morphology could be controlled by the relative contents of Mo and Ag. The Mo-Ag film has better hydrophilicity than the pure Ag film, which could be beneficial for cell attachment during biological SERS sensing. Moreover, the cell test shows that the Mo-Ag film exhibits good cytocompatibility with MC3T3-E1 cells, which shows potential for SERS detection in vivo. The high surface roughness of the Mo-Ag film is a fascinating feature for enhancing Raman scattering signals. Through depositing a thin film of Mo-Ag on a glass surface with a size of 0.5 × 0.5 cm2, a sensing chip of SERS could be produced. High sensitivity and excellent signal reproducibility were acquired. The sensitivity was down to 10-10 mol L-1 for malachite green and 10-9 mol L-1 for thiram, and the relative standard deviation value was lower than 7.0%. Moreover, bacteria were detected by employing the Mo-Ag film chip, and the difference in signal uniformity between molecules and bacteria is illustrated. In summary, depositing the Mo-Ag film on the surface of sensors could be an effective strategy for biomedical SERS sensing.

An integrated magneto-opto-fluidic biosensor for rapid on-chip assay of respiratory viruses of livestock.

Respiratory disease is one of the most important causes of economic loss in swine production. In the USA, porcine reproductive and respiratory syndrome virus (PRRSV) and influenza A virus (IAV) are currently the top two primary viruses causing swine respiratory diseases. The commonly used PCR-based virus detection methods require virus extraction, nucleic acid purification, and detection, which are relatively time-consuming and expensive. This work reports an integrated magneto-opto-fluidic (iMOF) platform, in which antibody functionalized magnetic nanoparticles (MNPs) can enable efficient enrichment of multiple swine respiratory viruses and a photonic crystal (PC) biosensor can transduce the amount of captured MNP-virus nanoparticles to the change of their reflection signatures. Owing to the high refractive index of Fe2O3 MNPs, the use of MNPs can significantly enhance the PC sensor output. The proof-of-concept validation involves using antibody-functionalized MNPs to recognize IAV and PRRSV and transferring the formed MNP-virus conjugates onto the surface of the PC biosensors to quantify these viruses. The iMOF platform offers a high sensitivity of 3.5 TCID50 mL-1 and 5.9 TCID50 mL-1 for detecting IAV and PRRSV, respectively, and a rapid turnaround within one hour, including the MNP-virus conjugation, enrichment, and detection. The on-chip virus platform has a great potential for in-field surveillance of viral infections.

Customized peptidoglycan surfaces to investigate innate immune recognition via surface plasmon resonance.

Glycan-protein interactions facilitate some of the most important biomolecular processes in and between cells. They are involved in different cellular pathways, cell-cell interactions and associated with many diseases, making these interactions of great interest. However, their structural and functional diversity poses great challenges in studying them at the molecular level. Surface plasmon resonance (SPR) technology presents great advantages to study glycan-protein interactions due to its superior sensitivity, ability to monitor real-time interactions, relatively simple data interpretation, and most importantly, direct measurement of binding without a need for fluorescent labeling. Here, another dimensionality of SPR in studying glycan-protein interactions is demonstrated via examples of binding between human innate immune receptors and their bacterial peptidoglycan ligands. In order to best resemble interactions in solution, a novel strategy of tethering the carbohydrate at different positions to the biosensor surface is applied to represent the potential displays of the carbohydrate ligand to the receptor. Subsequent kinetic analysis provides insights into the optimized configuration of peptidoglycan fragments for binding with its receptors. The manuscript contains a "how-to guide" to help with the implementation of these methods in other glycan-protein binding systems.

Copolymer Coatings for DNA Biosensors: Effect of Charges and Immobilization Chemistries on Yield, Strength and Kinetics of Hybridization

The physical–chemical properties of the surface of DNA microarrays and biosensors play a fundamental role in their performance, affecting the signal’s amplitude and the strength and kinetics of binding. We studied how the interaction parameters vary for hybridization of complementary 23-mer DNA, when the probe strands are immobilized on different copolymers, which coat the surface of an optical, label-free biosensor. Copolymers of N, N-dimethylacrylamide bringing either a different type or density of sites for covalent immobilization of DNA probes, or different backbone charges, were used to functionalize the surface of a Reflective Phantom Interface multispot biosensor made of a glass prism with a silicon dioxide antireflective layer. By analyzing the kinetic hybridization curves at different probe surface densities and target concentrations in solution, we found that all the tested coatings displayed a common association kinetics of about 9 × 104 M−1·s−1 at small probe density, decreasing by one order of magnitude close to the surface saturation of probes. In contrast, both the yield of hybridization and the dissociation kinetics, and hence the equilibrium constant, depend on the type of copolymer coating. Nearly doubled signal amplitudes, although equilibrium dissociation constant was as large as 4 nM, were obtained by immobilizing the probe via click chemistry, whereas amine-based immobilization combined with passivation with diamine carrying positive charges granted much slower dissociation kinetics, yielding an equilibrium dissociation constant as low as 0.5 nM. These results offer quantitative criteria for an optimal selection of surface copolymer coatings, depending on the application.

Fabrication and Application of CuO/ZnO/Rgo Nanocomposite Modified Pencil Graphite Electrodes for Urea and Uric Acid Biosensing in Uremic Samples

Gout is a disease associated with defective metabolism of uric acid that also results in arthritis with chalkstone deposits on bones of the feet, causing acute painful episodes. Both gout and hyperuricemia are diseases linked with certain levels of uric acid in suffers and even some patients with renal diseases. With the relationship between serum uric acid and urea as biometabolites, it was necessary to examine their uremic contents as contributive factors to chronic gout. This study was necessary since early detection of urea and uric acid, even at very minute levels, in human blood may foster a better understanding of renal health, facilitate prompt gout diagnosis, and eventually promote healthy living. In the present study, CuO/ZnO/reduced graphene oxide (rGO) nanocomposite modified pencil graphite electrodes are fabricated and appropriately characterized using surface morphological techniques (e.g., atomic force microscopy and scanning electron microscopy) and electrochemical techniques (e.g., cyclic voltammetry and electrochemical impedance spectroscopy). These nanocomposites sensors acted as substrates for adsorbing and detecting of urea and uric acid. Results from this study revealed a fabrication process for making modified electrode-based sensors for effective and simultaneous detection of both biomarkers at exceptionally low limit of detection (LOD) in uremic samples. No matrix interferences were recorded in both plasma and serum test samples. The optimum concentrations of these nanocomposites on the biosensor surfaces significantly recorded detection limits at 1.80 × 10-9 M for urea and 2.69× 10-9 M for uric acid in plasma while levels of urea and uric acid stood at 2.06 × 10-9 and 3.10× 10-9 M, respectively, in serum samples. The efficacy of this biomarker detection protocol was also examined from pharmacokinetic studies for both substances in their standard solutions and spiked uremic samples. Prompt sensing of these kinds of metabolites as disease biomarkers at low LOD is important in the assessment of degrading health conditions of gout patients as well as in monitoring inherent concentration changes for potential treatments. Furthermore, this detection protocol may have a future in bioassay involving gout diagnosis in renal patients, assessment of gross renal health or even in-vitro and in-vivo applications for monitoring drug dosages. Keywords: Gout, Biosensing; Uric acid; Simultaneous detection; Disease biomarker; Limit of detection

Nanoparticle-Based FM-MCF LSPR Biosensor With Open Air-Hole

A nanoparticle-based few-mode multi-core fiber (FM-MCF) localized surface plasmon resonance (LSPR) biosensor is proposed and analyzed using the finite element method (FEM). It’s critical to narrow the loss spectrum and improve the coupling efficiency, which makes it have high resolution and high sensitivity. With the aid of open air holes, the gold nanoparticles are easily assembled on the surface of this FM-MCF LSPR biosensor. Through multiple investigations, the performance of the sensor can be improved by properly setting gold nanoparticle configurations, such as radius, positions, shapes, and nanoparticle arrays. The simulation results show that when three circular gold nanoparticles with a radius of 150 nm are placed symmetrically in the open air hole and the angle between adjacent nanoparticles is 5°, the maximum sensitivity of 7,351.6 nm/RIU (LP02y mode na = 1.38) can be obtained in the sensing range of 1.33–1.38, which covers the refractive index (RI) of biological fluids, such as bovine serum albumin (BSA) solution and human Immunoglobulin G.

Design and Optimization of a Biosensor Surface Functionalization to Effectively Capture Urinary Extracellular Vesicles.

For this study, we tested and optimized silicon surface functionalization procedures for capturing urinary extracellular vesicles (uEVs). The influence of the silane type (APTES or GOPS) and protein concentration on the efficiency of uEVs binding was investigated. Human lactadherin protein (LACT) was used to capture uEVs. We applied surface characterization techniques, including ellipsometry, atomic force microscopy, and time-of-flight secondary ion mass spectrometry, to observe changes in the biosensor surface after each functionalization step. uEVs were purified by a low-vacuum filtration method and concentrated by ultracentrifugation. The physical parameters of uEVs after the isolation procedure, such as morphology and size distribution, were determined using transmission electron microscopy and tunable resistive pulse sensing methods. We observed a gradual growth of the molecular layer after subsequent stages of modification of the silicon surface. The ToF-SIMS results showed no changes in the mean intensities for the characteristic peaks of amino acids and lipids in positive and negative polarization, in terms of the surface-modifying silane (APTES or GOPS) used. The most optimal concentration of LACT for the tested system was 25 µg/mL.

Primary Human Renal Proximal Tubular Epithelial Cells (pHRPTEpiCs): Shiga Toxin (Stx) Glycosphingolipid Receptors, Stx Susceptibility, and Interaction with Membrane Microdomains

Tubular epithelial cells of the human kidney are considered as targets of Shiga toxins (Stxs) in the Stx-mediated pathogenesis of hemolytic–uremic syndrome (HUS) caused by Stx-releasing enterohemorrhagic Escherichia coli (EHEC). Analysis of Stx-binding glycosphingolipids (GSLs) of primary human renal proximal tubular epithelial cells (pHRPTEpiCs) yielded globotriaosylceramide (Gb3Cer) and globotetraosylceramide (Gb4Cer) with Cer (d18:1, C16:0), Cer (d18:1, C22:0), and Cer (d18:1, C24:1/C24:0) as the dominant lipoforms. Investigation of detergent-resistant membranes (DRMs) and nonDRMs, serving as equivalents for the liquid-ordered and liquid-disordered membrane phase, respectively, revealed the prevalence of Gb3Cer and Gb4Cer together with cholesterol and sphingomyelin in DRMs, suggesting lipid raft association. Stx1a and Stx2a exerted strong cellular damage with half-maximal cytotoxic doses (CD50) of 1.31 × 102 pg/mL and 1.66 × 103 pg/mL, respectively, indicating one order of magnitude higher cellular cytotoxicity of Stx1a. Surface acoustic wave (SAW) real-time interaction analysis using biosensor surfaces coated with DRM or nonDRM fractions gave stronger binding capability of Stx1a versus Stx2a that correlated with the lower cytotoxicity of Stx2a. Our study underlines the substantial role of proximal tubular epithelial cells of the human kidney being associated with the development of Stx-mediated HUS at least for Stx1a, while the impact of Stx2a remains somewhat ambiguous.