Transduction Techniques Research Articles

Electrochemiluminescence Lateral Flow Immunoassay Using Ruthenium(II) Complex‐Loaded Dendritic Mesoporous Silica Nanospheres for Highly Sensitive and Quantitative Detection of SARS‐CoV‐2 Nucleocapsid Protein

AbstractLateral flow immunoassays (LFIA) are widely used for the cost‐effective and rapid detection of diverse analytes. However, traditional LFIA suffers from difficulties in providing quantitative results and has low sensitivity. Herein, LFIA is combined with electrochemiluminescence (ECL), a leading transduction technique with high sensitivity and wide dynamic range, to achieve highly sensitive and quantitative detection of severe acute respiratory syndrome coronavirus nucleocapsid protein (SARS‐CoV‐2 N protein). Ruthenium(II)‐based complexes are synthesized and loaded into dendritic mesoporous silica nanospheres (PEI‐Ru/dSiO2), which possessed central‐radial pore channels and served as tags for ECL‐LFIA. The electrodes are fabricated on a nitrocellulose (NC) membrane, which simplifies the structure of the ECL‐LFIA. PEI‐Ru/dSiO2 is captured on the electrode surface via a sandwich immunoreaction, which enhances the ECL signal by decreasing the distance between PEI‐Ru/dSiO2 and the electrode surface. Using 2,2‐bis(hydroxymethyl)‐2,2′,2′'‐nitrilotriethanol (BIS‐TRIS) as coreactant, the ECL‐LFIA is used for detecting SARS‐CoV‐2 N protein, with a linear range of 1–104 ng mL−1 and a limit of detection (LOD) of 0.52 ng mL−1. ECL‐LFIA can also be used to detect analytes in complex matrices. These results demonstrate that the prepared ECL‐LFIA has great potential as a point‐of‐care testing platform for the rapid and quantitative detection of disease biomarkers.

Recent Advancements in Biosensors for the Detection and Characterization of Amyloids: A Review.

Modern medicine has increased the human lifespan. However, with an increase in average lifespan risk of amyloidosis increases. Amyloidosis is a condition characterized by protein misfolding and aggregation. Early detection of amyloidosis is crucial, yet conventional diagnostic methods are costly and lack precision, necessitating innovative tools. This review explores recent advancements in diverse amyloid detection methodologies, highlighting the need for interdisciplinary research to develop a miniaturized electrochemical biosensor leveraging nanotechnology. However, the diagnostics industry faces obstacles such as skilled labor shortages, standardized selection processes, and concurrent multi-analyte identification challenges. Research efforts are focused on integrating electrochemical techniques into clinical applications and diagnostics, with the successful transition of miniaturized technologies from development to testing posing a significant hurdle. Label-free transduction techniques like voltammetry and electrochemical impedance spectroscopy (EIS) have gained traction due to their rapid, cost-effective, and user-friendly nature.

Chronocoulometric Quantum Dot Aptasensor for SARS‐CoV‐2 Nucleocapsid Protein Biomarker

AbstractFor the first time, a novel mercaptosuccinic acid‐capped tungsten telluride quantum dot (MSA‐WTe3‐QD)‐based electrochemical aptasensor was designed for the determination of SARS‐CoV‐2 N‐protein in synthetic human serum, plasma and urine. The MSA‐WTe3‐QD which exhibited spherical morphology and an average diameter of 3.56 nm were synthesized through a bottom‐up approach. The aptasensor was prepared by firstly immobilizing MSA‐WTe3‐QD on a screen‐printed carbon electrode (SPCE) by using the 1‐ethyl‐3‐(3‐dimethylaminopropyl)carbodiimide/N‐hydroxysuccinimide (EDC/NHS) coupling chemistry. Thereafter, amine‐terminated 90‐mer ssDNA aptamer was deposited on the modified SPCE and left to cure to form the aptasensor. Using the chronocoulometry signal transduction technique, the aptasensor's calibration with N‐protein gave a linear range value of 1–9 pg.mL−1 and a limit of detection (LOD) value of 0.08 pg.mL−1. The LOD value of the aptasensor was 8 times lower than the LOD value of 0.71 pg.mL−1 obtained with a commercial N‐protein ELISA kit. When tested in simulated real samples the aptasensor recovery rates of 95–103.33 % were obtained. In the presence of various interferents, the aptasensor displayed good selectivity to N‐protein.

Progress on Electrochemical Biomimetic Nanosensors for the Detection and Monitoring of Mycotoxins and Pesticides.

Monitoring agricultural toxins such as mycotoxins is crucial for a healthy society. High concentrations of these toxins lead to the cause of several chronic diseases; therefore, developing analytical systems for detecting/monitoring agricultural toxins is essential. These toxins are found in crops such as vegetables, fruits, food, and beverage products. Currently, screening of these toxins is mostly performed with sophisticated instrumentation such as chromatography and spectroscopy techniques. However, these techniques are very expensive and require extensive maintenance, and their availability is limited to metro cities only. Alternatively, electrochemical biomimetic sensing methodologies have progressed hugely during the last decade due to their unique advantages like point-of-care sensing, miniaturized instrumentations, and mobile/personalized monitoring systems. Specifically, affinity-based sensing strategies including immunosensors, aptasensors, and molecular imprinted polymers offer tremendous sensitivity, selectivity, and stability to the sensing system. The current review discusses the principal mechanisms and the recent developments in affinity-based sensing methodologies for the detection and continuous monitoring of mycotoxins and pesticides. The core discussion has mainly focused on the fabrication protocols, advantages, and disadvantages of affinity-based sensing systems and different exploited electrochemical transduction techniques.

Enabling Non-invasive Tracking of Vascular Endothelial Cells Derived from Induced Pluripotent Stem Cells Using Nuclear Imaging.

The absence of clinically applicable imaging techniques for continuous monitoring of transplanted cells poses a significant obstacle to the clinical translation of stem cell-based therapies for vascular regeneration. This study aims to optimize a clinically applicable, non-invasive imaging technique to longitudinally monitor vascular endothelial cells (ECs) for vascular regeneration in peripheral artery disease (PAD). Human induced pluripotent stem cells (HiPSCs) were employed to generate ECs (HiPSC-ECs). Lentiviral vectors encoding human sodium iodide symporter (hNIS) and enhanced green fluorescent protein (eGFP) genes were introduced to HiPSCs and HiPSC-ECs at varying multiplicities of infection (MOI). Through a combination of fluorescence microscopy and flow cytometry, an optimized transduction technique for introducing hNIS-eGFP into HiPSC-ECs was established. Subsequently, single-photon emission computed tomography (SPECT) was utilized for imaging of the transduced cells in vitro and in vivo after transplantation into the gastrocnemius muscle of nude mice. Lentiviral transduction resulted in sustained co-expression of hNIS and eGFP in HiPSC-ECs when transduced post-endothelial differentiation. An optimal MOI of five yielded over 90% hNIS-eGFP expression efficiency without compromising cell viability. hNIS-eGFP+ HiPSC-ECs exhibited 99mTc uptake and were detectable through SPECT in vitro. Additionally, intramuscular injection of hNIS-eGFP+ HiPSC-ECs with MatrigelTM into the hindlimbs of nude mice enabled real-time SPECT/CT tracking, from which a reduction in signal exceeding 80% was observed within 7days. This study establishes an optimized cell modification and imaging protocol for tracking transplanted cells. Future efforts will focus on enhancing cell survival and integration via improved delivery systems, thereby advancing the potential of cell-based therapies for PAD.

Better Reception, Better Signal! Investigating the Impact of Electrochemical Immunosensor Architecture on Sensitivity

Enhancing digital healthcare monitoring to allow for the detection of biological species such as viruses, hormones, and early warning disease biomarkers requires sensors with ultra-low limits of detection, a broad linear range of performance, and high selectivity. Furthermore, these requirements are needed when analysing for target analyte species present in low concentration levels, as well as, for example, when pursuing non-invasive detection applications in saliva- or sweat-based analyte solutions. These analyte solutions can be complex, containing a variety of non-specific species and molecules. Therefore, focusing on the sensor architecture, surface functionalization, and the transduction method is of great importance when addressing these complex solutions.[2] Maximising the available surface area of the transducing sensor facilitates greater numbers of binding events between the receptor and target analyte of interest being possible.[1] This in turn increases sensitivity for identifying and quantifying the target analyte distinct from other species present, or in low concentrations in the solution. Evaluating different geometries, supporting structures, and the materials in the sensor electrode’s design can determine which architecture, in combination with the transduction technique and immobilized molecules, can enhance the overall performance of the sensor and attain ultra-low levels of detection. Additionally, while reliability in the bio-functionalization of the electrode surface is crucial, it is equally vital that the method of fabricating the electrodes geometry is repeatable. Repeatability in this regard further decreases sensor to sensor variability and signal outputs, especially if the sensor is to become commercially viable. In this work, custom micropillar-array silicon (Si) chips (MASCs) are commercially fabricated, thus providing the advantage of having a highly repeatable architecture and being commercially viable as a sensing design. The MASCs were used as a supporting structure to increase the available surface area, compared to a planar chip, for immobilising biorecognition elements, therefore increasing the sensitivity to potential binding events between the receptor and target analyte. The MASCs were coated with gold (Au) to facilitate the supporting surface acting as a transducing sensor, utilizing electrochemical impedance spectroscopy (EIS) as the primary transduction technique. The study evaluates the sensitivity and linear range of the MASCs comparatively against planar Au-coated Si chips, characterizing the design and performance of the sensors via EIS, cyclic voltammetry (CV), differential pulse voltammetry (DPV), fluorescent microscopy (FM), and scanning electron microscopy (SEM). With this study, a label-free transduction technique was harnessed for biosensing, addressing performance issues by developing different sensor architectures, leveraging a robust biofunctionalization protocol, and comparatively developing sensors whose design considerations accommodate goals of commercial viability and a robust, adaptable biosensing platform for future research prospectives.

Nanoparticles in Electrochemical Immunosensors – A Concept and Perspective

AbstractNanoparticle‐based electrochemical immunosensors are fascinating and promising screening systems that rely on the affinity binding between antibodies and antigens, allowing the specific detection of diverse types of targets (e. g., proteins, hormones, antibodies). The employment of nanoparticles in the realization of such devices dramatically improves the selectivity, sensitivity, and the transducer surface area for effectively immobilizing the bioreceptor or target molecules. During the last years, the nanoparticles design and fabrication was mainly focused on taking advantage from synergistic effects due to the coupling of different materials. In this review, we summarize the very recent advancements in this regard, with specific emphasis to nanoparticle syntheses procedures, antibody oriented immobilization strategies and transduction techniques. Moreover, the last trends and applications have been extensively discussed along with the remaining challenges and future perspectives.

Trends and challenges in electroanalytical biosensing methodologies for infectious viral diseases

Viral pandemic diseases have disruptive global consequences leading to millions of deaths and a severe impact on the global economy. Inadequate preventative protocols have led to an overwhelming demand for intensive care leading to uncontrollable burdens and even breakdown of healthcare sectors across many countries. The rapid detection of viral disease helps in the understanding of the relevant intricacies, helping to tackle infection with improved guidelines. Portable biosensor devices offer promising solutions by facilitating on-site detection of viral pathogens. This review summarizes the latest innovative strategies reported using electroanalytical methods for the screening of viral antigens. The structural components of viruses and their categories are presented followed by the various recognition elements and transduction techniques involved in biosensors. Core sections focus on biosensors reported for viral genomic detection(DNA and RNA) and antigenic capsid protein. Strategies for addressing the challenges of electroanalytical biosensing of viral components are also presented. The advantages, and disadvantages of biorecognition elements and nanozymes for the detection of viral disease are highlighted. Such technical insights will help researchers working in chemistry, and biochemistry as well as clinicians working in medical diagnostics.

Carbon Electrode Modified with Molecularly Imprinted Polymers for the Development of Electrochemical Sensor: Application to Pharmacy, Food Safety, Environmental Monitoring, and Biomedical Analysis

This review aims to elucidate recent developments in electrochemical sensors that use functionalized carbon electrodes with molecularly imprinted polymers (MIPs) for the selective detection of organic compounds in diverse fields including pharmacy, food safety, environmental monitoring of pollutants, and biomedical analysis. The main targets include explosive compounds, dyes, antioxidants, disease biomarkers, pharmaceuticals, antibiotics, allergens, pesticides, and viruses. Following a brief overview of the molecular imprinting principle, the most significant applications are explored. The selection of the functional monomer is subsequently discussed. Notably, various types of carbon electrodes are presented, with a particular emphasis on screen-printed carbon electrodes. The most commonly employed techniques for MIP deposition such as electropolymerization, drop casting, and chemical grafting are introduced and discussed. Electrochemical transduction techniques like cyclic voltammetry, differential pulse voltammetry, square wave voltammetry, and electrochemical impedance spectroscopy are presented. Lastly, the review concludes by examining potential future directions and primary limitations concerning carbon electrodes modified with MIPs.

Activated carbon derived from wood biochar for Amperometric sensing of Ammonia for early screening of chronic kidney disease

Personalized medicine has emerged as an increasingly efficient and effective approach to addressing disease diagnosis and intervention. Ammonia is a waste product produced by the body during the digestion of protein. The requirement to develop an electrochemical sensing platform for monitoring skin ammonia levels holds great potential as an essential solution to pre-screen chronic kidney disease (CKD). In this research, we have manufactured an innovative electrochemical sensor by employing activated carbon derived from wood biochar as the signal transducer. We conducted a comprehensive analysis of the structural and morphological characteristics of the synthesized materials using various techniques. The hypothesized interaction was investigated using chronoamperometry as a transduction technique. To assess cross-reactivity, we conducted a study using common interferants or chemicals present in the environment. The data presented in this paper represents three replicates and is plotted with a 5 % error bar, demonstrating a 95 % confidence interval in the sensor response. In this study, we have elucidated the functionality and usefulness of a wearable microelectronic research prototype integrated with an HTC-activated carbon @RTIL-based electrochemical sensing platform for detecting ammonia levels released from the skin as a marker for chronic kidney disease screening. By enabling early detection and monitoring, these platforms can facilitate timely interventions, such as lifestyle modifications, medication adjustments, or referral to nephrology specialists. This proactive approach can potentially slow down disease progression, minimize the need for dialysis or transplantation, and ultimately improve the quality of life for CKD patients.

Optical Immunosensors for Bacteria Detection in Food Matrices

Optical immunosensors are one of the most popular categories of immunosensors with applications in many fields including diagnostics and environmental and food analysis. The latter field is of particular interest not only for scientists but also for regulatory authorities and the public since food is essential for life but can also be the source of many health problems. In this context, the current review aims to provide an overview of the different types of optical immunosensors focusing on their application for the determination of pathogenic bacteria in food samples. The optical immunosensors discussed include sensors based on evanescent wave transduction principles including surface plasmon resonance (SPR), fiber-optic-, interferometric-, grating-coupler-, and ring-resonator-based sensors, as well as reflectometric, photoluminescence, and immunosensors based on surface-enhanced Raman scattering (SERS). Thus, after a short description of each transduction technique, its implementation for the immunochemical determination of bacteria is discussed. Finally, a short commentary about the future trends in optical immunosensors for food safety applications is provided.

Reticular Electrochemiluminescence Nanoemitters: Structural Design and Enhancement Mechanism.

ConspectusElectrochemiluminescence (ECL) is a powerful transduction technique, which depends critically on the formation of the excited emitter through the charge transfer between the electrochemical reaction intermediates of the emitter and the co-reactant/emitter. The exploration of ECL mechanisms for conventional nanoemitters is limited due to the uncontrollable charge transfer process. With the development of molecular nanocrystals, reticular structures such as metal-organic frameworks (MOFs) and covalent organic frameworks (COFs) have been utilized as atomically precise semiconducting materials. The long-range order in crystalline frameworks and the tunable coupling among building blocks promote the quick development of electrically conductive frameworks. Especially, the reticular charge transfer can be regulated by both interlayer electron coupling and intralayer topology-templated conjugation. By modulating intramolecular or intermolecular charge mobility, reticular structures could serve as promising candidates for enhancing ECL. Thus, reticular crystalline nanoemitters with different topologies provide a confined platform to understand ECL fundamentals for designing next-generation ECL devices.Aiming at exploring the mechanism of ECL emission, our group has developed a series of ECL nanoemitters as well as enhancement strategies of ECL emission in the past 20 years. A series of water-soluble ligand-capped quantum dots were introduced as ECL nanoemitters to create sensitive analytical methods for detecting and tracing biomarkers. The functionalized polymer dots were also designed as ECL nanoemitters for imaging of membrane proteins with signal transduction strategies of dual resonance energy transfer and dual intramolecular electron transfer. To decode the ECL fundamental and enhancement mechanisms, an electroactive MOF with accurate molecular structure was first constructed with two redox ligands as a highly crystallized ECL nanoemitter in aqueous medium. Through the mixed-ligand approach, luminophores and co-reactants were integrated into one MOF structure for self-enhanced ECL. Furthermore, several donor-acceptor COFs were developed as efficient ECL nanoemitters with tunable intrareticular charge transfer. The atomically precise structure of conductive frameworks established clear correlations between the structure and charge transport in these materials. Therefore, reticular materials as crystalline ECL nanoemitters have demonstrated both proof of concept and mechanistic innovation.In this Account, taking advantage of reticular materials with accurate molecular structure, we survey the design of the electroactive reticular materials including MOFs and COFs as crystalline ECL nanoemitters at the molecular level. The enhancement mechanisms of ECL emission of various topology frameworks are discussed via the regulation of reticular energy transfer and charge transfer and the accumulation of anion/cation radicals. Our perspective on the reticular ECL nanoemitters is also discussed. This Account provides a new avenue for designing molecular crystalline ECL nanoemitters and decoding the fundamentals of ECL detection methods.

Enzymatic Catalysis in Size and Volume Dual-Confined Space of Integrated Nanochannel-Electrodes Chip for Enhanced Impedance Detection of Salmonella.

Nanochannel-based confinement effect is a fascinating signal transduction strategy for high-performance sensing, but only size confinement is focused on while other confinement effects are unexplored. Here, a highly integrated nanochannel-electrodes chip (INEC) is created and a size/volume-dual-confinement enzyme catalysis model for rapid and sensitive bacteria detection is developed. The INEC, by directly sandwiching a nanochannel chip (60µm in thickness) in nanoporous gold layers, creates a micro-droplet-based confinement electrochemical cell (CEC). The size confinement of nanochannel promotes the urease catalysis efficiency to generate more ions, while the volume confinement of CEC significantly enriches ions by restricting diffusion. As a result, the INEC-based dual-confinement effects benefit a synergetic enhancement of the catalytic signal. A 11-times ion-strength-based impedance response is obtained within just 1min when compared to the relevant open system. Combining this novel nanoconfinement effects with nanofiltration of INEC, a separation/signal amplification-integrated sensing strategy is further developed for Salmonella typhimurium detection. The biosensor realizes facile, rapid (<20min), and specific signal readout with a detection limit of 9 CFU mL-1 in culturing solution, superior to most reports. This work may create a new paradigm for studying nanoconfined processes and contribute a new signal transduction technique for trace analysis application.

Cell therapy biomanufacturing: integrating biomaterial and flow-based membrane technologies for production of engineered T-cells.

Adoptive T-cell therapies (ATCTs) are increasingly important for the treatment of cancer, where patient immune cells are engineered to target and eradicate diseased cells. The biomanufacturing of ATCTs involves a series of time-intensive, lab-scale steps, including isolation, activation, genetic modification, and expansion of a patient's T-cells prior to achieving a final product. Innovative modular technologies are needed to produce cell therapies at improved scale and enhanced efficacy. In this work, well-defined, bioinspired soft materials were integrated within flow-based membrane devices for improving the activation and transduction of T cells. Hydrogel coated membranes (HCM) functionalized with cell-activating antibodies were produced as a tunable biomaterial for the activation of primary human T-cells. T-cell activation utilizing HCMs led to highly proliferative T-cells that expressed a memory phenotype. Further, transduction efficiency was improved by several fold over static conditions by using a tangential flow filtration (TFF) flow-cell, commonly used in the production of protein therapeutics, to transduce T-cells under flow. The combination of HCMs and TFF technology led to increased cell activation, proliferation, and transduction compared to current industrial biomanufacturing processes. The combined power of biomaterials with scalable flow-through transduction techniques provides future opportunities for improving the biomanufacturing of ATCTs.

Reversibly Tuning Electrochemiluminescence with Stimulated Emission Route for Single-Cell Imaging

Electrochemiluminescence (ECL) has established itself as an excellent transduction technique in biosensing and light-emitting device, while conventional ECL mechanism depending on spontaneous emission of luminophores lacks reversibility and tunable emission characters, limiting the universality of ECL technique in the fields of fundamental research and clinical applications. Here, we report the first observation of stimulated emission route in ECL and thus establish a reversible tuning ECL microscopy for single-cell imaging. This microscopy uses a focused red-shifted beam to transfer spontaneous ECL into stimulated ECL, which enables selective and reversible tuning of ECL emission from homogeneous solution, single particles, and single cells. After excluding other possible competitive routes, the stimulated ECL emission route is confirmed by a dual-objective system in which the suppressed spontaneous ECL is accompanied by the enhanced stimulated ECL. By incorporating a commercial donut-shaped beam, the sharpness of single-cell matrix adhesion is improved 2 to 3 times compared with the counterpart in confocal ECL mode. The successful establishment of this stimulated emission ECL will greatly advance the development of light-emitting device and super-resolution ECL microscopy.

Exploiting epoxy-rich poly(ethylene glycol) films for highly selective ssDNA sensing via electrochemical impedance spectroscopy.

This study introduces an alternative approach to immobilize thiolated single-stranded DNA (ssDNA) for the DNA sensing. In contrast to the standard, monomolecular assembly of such moieties on gold substrate, over the thiolate-gold anchors, we propose to use bioinert, porous polyethylene glycol (PEG) films as a 3D template for ssDNA immobilization. The latter process relies on the reaction between the thiol group of the respectively decorated ssDNA and the epoxy groups in the epoxy-rich PEG matrix. The immobilization process and subsequent hybridization ability of the resulting sensing assembly were monitored using cyclic voltammetry and electrochemical impedance spectroscopy, with the latter tool proving itself as the most suitable transduction technique. Electrochemical data confirmed the successful immobilization of thiol-decorated ssDNA probes into the PEG matrix over the thiol-epoxy linkage as well as high hybridization efficiency, selectivity, and sensitivity of the resulting DNA sensor. Whereas this sensor was equivalent to the direct ssDNA assembly in terms of the efficiency, it exhibited a better selectivity and bioinert properties in view of the bioinert character of the PEG matrix. The above findings place PEG films as a promising platform for highly selective ssDNA sensing, leveraging their flexible chemistry, 3D character, and bioinert properties.

Selective Antibody-Free Sensing Membranes for Picogram Tetracycline Detection.

As an antibody-free sensing membrane for the detection of the antibiotic tetracycline (TC), a liquid PVC membrane doped with the ion-pair tetracycline/θ-shaped anion [3,3'-Co(1,2-C2B9H11)2]- ([o-COSAN]-) was formulated and deposited on a SWCNT modified gold microelectrode. The chosen transduction technique was electrochemical impedance spectroscopy (EIS). The PVC membrane was composed of: the tetracycline/[o-COSAN]- ion-pair, a plasticizer. A detection limit of 0.3 pg/L was obtained with this membrane, using bis(2-ethylhexyl) sebacate as a plasticizer. The sensitivity of detection of tetracycline was five times higher than that of oxytetracycline and of terramycin, and 22 times higher than that of demeclocycline. A shelf-life of the prepared sensor was more than six months and was used for detection in spiked honey samples. These results open the way to having continuous monitoring sensors with a high detection capacity, are easy to clean, avoid the use of antibodies, and produce a direct measurement.

Emerging Biosensing Technologies towards Early Sepsis Diagnosis and Management.

Sepsis is defined as a systemic inflammatory dysfunction strictly associated with infectious diseases, which represents an important health issue whose incidence is continuously increasing worldwide. Nowadays, sepsis is considered as one of the main causes of death that mainly affects critically ill patients in clinical settings, with a higher prevalence in low-income countries. Currently, sepsis management still represents an important challenge, since the use of traditional techniques for the diagnosis does not provide a rapid response, which is crucial for an effective infection management. Biosensing systems represent a valid alternative due to their characteristics such as low cost, portability, low response time, ease of use and suitability for point of care/need applications. This review provides an overview of the infectious agents associated with the development of sepsis and the host biomarkers suitable for diagnosis and prognosis. Special focus is given to the new emerging biosensing technologies using electrochemical and optical transduction techniques for sepsis diagnosis and management.

Rational Design of Porous Poly(ethylene glycol) Films as a Matrix for ssDNA Immobilization and Hybridization.

Poly(ethylene glycol) (PEG) films, fabricated by thermally induced crosslinking of amine- and epoxy-terminated four-arm STAR-PEG precursors, were used as porous and bioinert matrix for single-stranded DNA (ssDNA) immobilization and hybridization. The immobilization relied on the reaction between the amine groups in the films and N-hydroxy succinimide (NHS) ester groups of the NHS-ester-decorated ssDNA. Whereas the amount of reactive amine groups in the films with the standard 1:1 composition of the precursors turned out to be too low for efficient immobilization, it could be increased noticeably using an excess (2:1) concentration of the amine-terminated precursor. The respective films retained the bioinertness of the 1:1 prototype and could be successfully decorated with probe ssDNA, resulting in porous, 3D PEG-ssDNA sensing assemblies. These assemblies exhibited high selectivity with respect to the target ssDNA strands, with a hybridization efficiency of 78–89% for the matching sequences and full inertness for non-complementary strands. The respective strategy can be applied to the fabrication of DNA microarrays and DNA sensors. As a suitable transduction technique, requiring no ssDNA labeling and showing high sensitivity in the PEG-ssDNA case, electrochemical impedance spectroscopy is suggested.

Elucidating Sensitivity and Stability Relationship of Gold-Carbon Hybrid LSPR Sensors Using Principal Component Analysis.

Sensitive localized surface plasmon resonance (LSPR) sensing is achieved using nanostructured geometries of noble metals which typically have dimensions less than 100 nm. Among the plethora of geometries and materials, the spherical geometries of gold (Au) are widely used to develop sensitive bio/chemical sensors due to ease of manufacturing and biofunctionlization. One major limitation of spherical-shaped geometries of Au, used for LSPR sensing, is their low refractive index (RI) sensitivity which is commonly addressed by adding another material to the Au nanostructures. However, the process of addition of new material on Au nanostructures, while retaining the LSPR of Au, often comes with a trade-off which is associated with the instability of the developed composite, especially in harsh chemical environments. Addressing this challenge, we develop a Au-graphene-layered hybrid (Au-G) with high stability (studied up to 2 weeks here) and enhanced RI sensitivity (a maximum of 180.1 nm/RIU) for generic LSPR sensing applications using spherical Au nanostructures in harsh chemical environments, involving organic solvents. Additionally, by virtue of principal component analysis, we correlate stability and sensitivity of the developed system. The relationship suggests that the shelf life of the material is proportional to its sensitivity, while the stability of the sensor during the measurement in liquid environment decreases when the sensitivity of the material is increased. Though we uncover this relationship for the LSPR sensor, it remains evasive to explore similar relationships within other optical and electrochemical transduction techniques. Therefore, our work serves as a benchmark report in understanding/establishing new correlations between sensing parameters.