Biosensor Applications Research Articles

A review on development of enzymatic biosensors for industrial applications

Enzymatic biosensors are vital tools across various industries due to their high specificity, sensitivity, and rapid response times. They are extensively used in medical diagnostics for monitoring biomarkers like glucose and lactate, environmental monitoring to detect pollutants, food safety and quality assurance, bioprocess control in industrial fermentation, pharmaceutical drug development and quality control, soil health and agrochemical detection in agriculture, and for identifying biological warfare agents. However, several challenges limit their broader adoption and effectiveness in industrial settings. Key challenges include enzyme stability, as enzymes can denature under extreme environmental conditions, reducing sensor lifespan and performance. Variability in enzyme activity and immobilization techniques also leads to inconsistencies, affecting sensor reproducibility and reliability. Complex sample matrices, such as those in environmental and food samples, introduce interference that can affect sensor accuracy. Integrating biosensors into existing industrial processes is another significant challenge, requiring compatibility with automated systems and real-time data processing capabilities. Additionally, high production costs and scalability issues hinder widespread adoption. Regulatory and standardization barriers further complicate the development and deployment of enzymatic biosensors. Despite these limitations, advancements in materials science, nanotechnology, and bioengineering are paving the way for more robust and reliable biosensors. Innovations such as nanomaterial-based enzyme immobilization, synthetic biology for enzyme engineering, and advanced data analytics integration hold promise for overcoming current limitations. Collaborative efforts among researchers, industry stakeholders, and regulatory bodies are essential to accelerate the development and application of enzymatic biosensors in industrial settings. While enzymatic biosensors offer significant potential, addressing these challenges through ongoing research and technological advancements is crucial for their widespread implementation and optimization.

Biosensing and Biotechnological Applications of Nanofillers: Current Status and Perspectives

Biosensing and Biotechnological Applications of Nanofillers: Current Status and Perspectives

Surface plasmon resonance sensors: Temperature effects

In this paper, we present a detailed study of the temperature effect (0−100°C), on the plasmonic resonance and sensitivity of a surface plasmon resonance (SPR) to the medium detection refraction index change. We examine this SPR based on the Kretschmann multilayer configuration constituted by a BK7 prism, an Ag/Au bimetallic layer deposited on TiO2/SiO2 and a BlueP/MoS2 heterostructure monolayer. First, a simple numerical calculation is provided to analyze the temperature effect on the plasmonic resonance. The model take into account the dependence on temperature of the refractive index of different materials composing the investigated structures. The main idea is to show how the sensitivity of the SPR based sensor is influenced according to operating temperature which is critical parameter when considering bio-sensing applications.

Redesigned Guide DNA Enhanced Clostridium butyricum Argonaute Activity for Amplification-Free and Multiplexed Detection of Pathogens.

Clostridium butyricum (CbAgo)-based bioassays are popular due to their programmability and directional cleavage capabilities. However, the relatively compact protein structure of CbAgo limits its cleavage activity (even at the optimal temperature), thus restricting its wider application. Here, we observed that guide DNA (gDNA) with specific structural features significantly enhanced CbAgo cleavage efficiency. Then, we invented a novel gDNA containing DNAzyme segments (gDNAzyme) that substantially enhanced the CbAgo cleavage efficency (by 100%). Using a molecular dynamics simulation system, we found that the augmented cleavage efficiency might be attributed to the large-scale global movement of the PIWI domain of CbAgo and an increased number of cleavage sites. Moreover, this gDNAzyme feature allowed us to create a biosensor that simultaneously and sensitively detected three pathogenic bacteria without DNA extraction and amplification. Our work not only dramatically expands applications of the CbAgo-based biosensor but also provides unique insight into the protein-DNA interactions.

Biosynthesis of Indigo Dyes and Their Application in Green Chemical and Visual Biosensing for Heavy Metals.

Fermentative production of natural colorants using microbial strains has emerged as a cost-effective and sustainable alternative to chemical synthesis. Visual pigments are used as signal outputs in colorimetric bacterial biosensors, a promising method for monitoring environmental pollutants. In this study, we engineered four self-sufficient indigo-forming enzymes, including HbpAv, bFMO, cFMO, and rFPMO, in a model bacterium E. coli. TrxA-bFMO was chosen for its strong ability to produce indigo under T7 lac and mer promoters' regulation. The choice of bacterial hosts, the supplementation of substrate l-tryptophan, and ventilation were crucial factors affecting indigo production. The indigo reporter validated the biosensors for Hg(II), Pb(II), As(III), and Cd(II). The biosensors reported Hg(II) as low as 14.1 nM, Pb(II) as low as 1.5 nM, and As(III) as low as 4.5 nM but increased to 25 μM for Cd(II). The detection ranges for Hg(II), Pb(II), As(III), and Cd(II) were quantified from 14.1 to 225 nM, 1.5 to 24.4 nM, 4.5 to 73.2 nM, and 25 to 200 μM, respectively. The sensitivity, responsive concentration range, and selectivity are comparable to β-galactosidase and luciferase reporter enzymes. This study suggests that engineered enzymes for indigo production have great potential for green chemical synthesis. Additionally, heterologous biosynthesis of indigo production can lead to the development of novel, low-cost, and mini-equipment bacterial biosensors with zero background noise for visual monitoring of pollutant heavy metals.

Mitigation of Device Heterogeneity in Graphene Hall Sensor Arrays Using Per-Element Backgate Tuning.

Graphene Hall-effect magnetic field sensors (GHSs) exhibit high performance comparable to state-of-the-art commercial Hall sensors made from III-V semiconductors. Graphene is also amenable to CMOS-compatible fabrication processes, making GHSs attractive candidates for implementing magnetic sensor arrays for imaging fields in biosensing and scanning probe applications. However, their practical appeal is limited by response heterogeneity and drift, arising from the high sensitivity of two-dimensional (2D) materials to local device imperfections. To address this challenge, we designed a GHS array in which an individual backgate is added to each GHS, allowing the carrier density of each sensor to be electrostatically tuned independent of other sensors in the array. Compared to the constraints encountered when all devices are tuned with the same backgate, we expected that the flexibility afforded by individual tuning would allow for the array's sensitivity, uniformity, and reconfigurability to be enhanced. We fabricated an array of 16 GHSs, each with its own backgate terminal, and characterized the ability to modulate GHS carrier density and Hall sensitivity within CMOS-compatible voltage ranges. We then demonstrated that individual device tuning can be used to break the trade-off between device sensitivity and uniformity in the GHS array, allowing for enhancement of both objectives. Our results showed that GHS arrays exhibiting >30% variability under single-backgate operation could be compensated using individual tuning to achieve <2% variability with minimal impact on the array sensitivity.

Optical Sensitive Detection of Cervical Cancer Based on Surface Plasmon Resonance Nanostructure

Cervical cancer is a disease that affects a lot of people all over the world. The success of treatment is significantly increased by early detection. The new surface plasmon resonance sensor (SPRS) described in this article is constructed from layers of silver, BaTiO3, and graphene, as well as a coupling prism. The transfer matrix (TM) approach is used to evaluate the SPRS structure. To obtain the highest sensitivity of the suggested SPRS, the thicknesses of the Ag and BaTiO3 and the number of Gr sheets are evaluated. Investigations are done on all factors of performance of the proposed device. Exemplary efficiency is achieved using 4 nm (BaTiO3) and 60 nm (Ag) materials. Three layers are chosen as the optimal amount of graphene after investigation. When the optimal thicknesses are employed, the suggested SPRS's greatest sensitivity of 300° RIU−1 has been attained. The suggested SPRS displays better sensitivity when compared to SPRS structures that have already been published in the literature. This SPRS is promising to be used in different biosensing applications due to its high performance.

Advances in Plasmonic Photonic Crystal Fiber Biosensors: Exploring Innovative Materials for Bioimaging and Environmental Monitoring

AbstractThis review paper comprehensively analyzes recent advancements in optical fiber‐based biosensors, focusing on conventional fiber and photonic crystal structures. This paper overviews the significant applications of optical fiber biosensors, including bioimaging, quality analysis, food safety, and field environment monitoring, setting the stage for subsequent discussions. The primary objective of the review is to systematically evaluate recent literature concerning optical fiber‐based biosensors, emphasizing their sensitivities and resolutions. The second section explores integrating plasmonic materials such as graphene, TDMC, germanium, black phosphorus, and silicon within optical fiber biosensors, elucidating their roles in enhancing sensitivity and resolution in biosensing applications. A detailed examination of photonic crystal fibers (PCF) follows, categorizing them into internally and externally metal film‐coated biosensors, highlighting their distinct advantages and limitations. Comparative analyses in two tables delineate the performance and sensitivity of optical fiber‐based biosensors, mainly focusing on different coating strategies. The final section of the review discusses emerging trends and applications in optical fiber biosensing technologies, underscoring their potential to transform biomedical and environmental monitoring fields. By synthesizing recent developments and challenges, this review aims to offer researchers and practitioners a comprehensive understanding of optical fiber‐based biosensors, facilitating informed decision‐making and driving further advancements in the field.

Unveiling Colorectal Cancer Biomarkers: Harnessing Biosensor Technology for Volatile Organic Compound Detection.

Conventional screening options for colorectal cancer (CRC) detection are mainly direct visualization and invasive methods including colonoscopy and flexible sigmoidoscopy, which must be performed in a clinical setting and may be linked to adverse effects for some patients. Non-invasive CRC diagnostic tests such as computed tomography colonography and stool tests are either too costly or less reliable than invasive ones. On the other hand, volatile organic compounds (VOCs) are potentially ideal non-invasive biomarkers for CRC detection and monitoring. The present review is a comprehensive presentation of the current state-of-the-art VOC-based CRC diagnostics, with a specific focus on recent advancements in biosensor design and application. Among them, breath-based chromatography pattern analysis and sampling techniques are overviewed, along with nanoparticle-based optical and electrochemical biosensor approaches. Limitations of the currently available technologies are also discussed with an outlook for improvement in combination with big data analytics and advanced instrumentation, as well as expanding the scope and specificity of CRC-related volatile biomarkers.

Impact of Air-Cathodes on Operational Stability of Single-Chamber Microbial Fuel Cell Biosensors for Wastewater Monitoring

The increasing global water pollution leads to the need for urgent development of rapid and accurate water quality monitoring methods. Microbial fuel cells (MFCs) have emerged as real-time biosensors for biochemical oxygen demand (BOD), but they grapple with several challenges, including issues related to reproducibility, operational stability, and cost-effectiveness. These challenges are substantially shaped by the selection of an appropriate air-breathing cathode. Previous studies indicated a critical influence of the cathode on both the enduring electrochemical performance of MFCs and the taxonomic diversity at the electroactive anode. However, the effect of different gas diffusion electrodes (GDE) on 3D-printed single-chamber MFCs for BOD biosensing application and its effect on the bioelectroactive anode was not investigated before. Our study focuses on comparing GDE cathode materials to enhance MFC performance for precise and rapid BOD analysis in wastewater. We examined for over 120 days two Pt-coated air-breathing cathodes with distinct carbonaceous gas diffusion layers (GDLs) and catalyst layers (CLs): cost-effective carbon paper (CP) with hand-coated CL and more expensive woven carbon cloth (CC) with CL pre-applied by the supplier. The results show significant differences in electrochemical characteristics and anodic biofilm composition between MFCs with CP and CC GDE cathodes. CP-MFCs exhibited lower sensitivity (16.6 C L mg−1 m−2) and a narrower dynamic range (25 to 600 mg L−1), attributed to biofouling-related degradation of the GDE. In contrast, CC-MFCs demonstrated superior performance with higher sensitivity (37.6 C L mg−1 m−2) and a broader dynamic range (25 to 800 mg L−1). In conclusion, our study underscores the pivotal role of cathode selection in 3D-printed MFC biosensors, influencing anodic biofilm enrichment time and overall BOD assessment performance. We recommend the use of cost-effective CP GDL with hand-coated CL for short-term MFC biosensor applications, while advocating for CC GDL supplied with CL as the preferred choice for long-term sensing implementations with enduring reliability.

Hybrid Mn Atomic Clusters/Single-Dispersed Atoms with Dual Antioxidant Activities for a Chemiluminescent Immunoassay.

Single-dispersed atoms (SDAs) as catalysts have drawn extensive attention due to their ultimate atom utilization efficiency and desirable catalytic capability. Atomic clusters (ACs) with potential multiple enzyme-like activities also display great practicability in catalysis-based biosensing. In this work, hybrid Mn ACs/SDAs were implanted in the frameworks of defect-engineered MIL 101(Cr) modulated by excess acetic acid, with a high loading capability of 13.9 wt %. Distinctively, Mn SDAs display weak superoxide dismutase (SOD)-like activity for specifically eliminating superoxide anion (O2•-), while Mn ACs/SDAs display both catalase-like and SOD-like activities for remarkable elimination of total reactive oxygen species (ROS) due to the cooperative effect of the two atom-scale catalytic sites. Thus, Mn ACs/SDAs can efficiently inhibit the chemiluminescent (CL) emission of multiple ROS-mediated luminol systems with a superior quenching rate of 85.5%. To validate the practicability of Mn ACs/SDAs for a sensitive CL assay, an immunoassay method was established to detect acetamiprid by using Mn ACs/SDAs as signal quenchers, which displayed a quantification range of 10 pg mL-1-25 ng mL-1 and a detection limit of 3.3 pg mL-1. This study paves an avenue for developing ACs/SDAs with multiple antioxidant activities that are suitable for application in biosensing.

Light-Powered Directional Ion Transport via PFN-Br/MoS2 Heterogeneous Membranes: Band Alignment and Activation Energy Barrier Engineering.

Biological photoresponsive ion transport systems consistently attract researchers' attention owing to their remarkable functions of harvesting energy from nature and participating in visual perception systems. Designing and constructing artificial light-driven ion transport devices to mimic biological counterparts remains a challenge owing to fabrication limitations in nanoconfined spaces. Herein, a typical conjugated polyelectrolyte (PFN-Br) was assembled onto a laminated MoS2M using simple solution-processing vacuum filtration, resulting in a heterogeneous three- and two-dimensional nanoporous membrane. The designed band alignment between PFN-Br and MoS2 enables effective directional ion transport under irradiation in an equilibrium solution, even against a 30-fold concentration gradient. The staggered energy structure of PFN-Br and MoS2 enhances charge separation and establishes a photogenerated potential as the driving force for ion transport. Additionally, the activation energy barrier for ion transport across the heterogeneous membrane decreased by 60% after light irradiation, considerably improving ion transport flux. The easy fabrication and high performance of the membrane in light-powered ion transport provide promising approaches for designing nanofluidic devices with possible applications in energy conversion, light-enhanced biosensing, and photoresponsive ionic devices.

Real-time monitoring of cellular superoxide anion release in THP-1 cells using a catalytically amplified superoxide dismutase-based microbiosensor.

Reactive oxygen species (ROS) including the superoxide anion (O2•-) are typically studied in cell cultures using fluorescent dyes, which provide only discrete single-point measurements. These methods lack the capabilities for assessing O2•- kinetics and release in a quantitative manner over long monitoring times. Herein, we present the fabrication and application of an electrochemical biosensor that enables real-time continuous monitoring of O2•- release in cell cultures for extended periods (> 8h) using an O2•- specific microelectrode. To achieve the sensitivity and selectivity requirements for cellular sensing, we developed a biohybrid system consisting of superoxide dismutase (SOD) and Ti3C2Tx MXenes, deposited on a gold microwire electrode (AuME) as O2•- specific materials with catalytic amplification through the synergistic action of the enzyme and the biomimetic MXenes-based structure. The biosensor demonstrated a sensitivity of 18.35 nA/μM with a linear range from 147 to 930nM in a cell culture medium. To demonstrate its robustness and practicality, we applied the biosensor to monitor O2•- levels in human leukemia monocytic THP-1 cells upon stimulation with lipopolysaccharide (LPS). Using this strategy, we successfully monitored LPS-induced O2•- in THP-1 cells, as well as the quenching effect induced by the ROS scavenger N-acetyl-L-cysteine (NAC). The biosensor is generally useful for exploring the role of oxidative stress and longitudinally monitoring O2•- release in cell cultures, enabling studies of biochemical processes and associated oxidative stress mechanisms in cellular and other biological environments.

Rationalizing Diverse Binding Mechanisms to the Same Protein Fold: Insights for Ligand Recognition and Biosensor Design.

The engineering of novel protein-ligand binding interactions, particularly for complex drug-like molecules, is an unsolved problem, which could enable many practical applications of protein biosensors. In this work, we analyzed two engineered biosensors, derived from the plant hormone sensor PYR1, to recognize either the agrochemical mandipropamid or the synthetic cannabinoid WIN55,212-2. Using a combination of quantitative deep mutational scanning experiments and molecular dynamics simulations, we demonstrated that mutations at common positions can promote protein-ligand shape complementarity and revealed prominent differences in the electrostatic networks needed to complement diverse ligands. MD simulations indicate that both PYR1 protein-ligand complexes bind a single conformer of their target ligand that is close to the lowest free-energy conformer. Computational design using a fixed conformer and rigid body orientation led to new WIN55,212-2 sensors with nanomolar limits of detection. This work reveals mechanisms by which the versatile PYR1 biosensor scaffold can bind diverse ligands. This work also provides computational methods to sample realistic ligand conformers and rigid body alignments that simplify the computational design of biosensors for novel ligands of interest.

Multiple Fano Resonance Modes based Plasmonic Refractive Index Sensor for Edible Oil Adulteration Detection

Surface plasmon polariton (SPP)-based plasmonic sensors are increasingly replacing traditional bulky sensors in refractive index sensing, biosensing, food adulteration detection, and other applications due to their unique optical features and different Metal-Insulator-Metal (MIM) topologies. The present research introduces a new design for a highly sensitive MIM waveguide-based refractive index sensor. This design incorporates an innovative combination of an inverted U-shaped rectangular split-ring resonator (RSRR), a semi-circle-split-ring resonator (SCSRR), and a stub. The sensor’s wave-guiding capabilities, such as electromagnetic wave transmission, field profile distribution, and sensitivity, are meticulously analyzed using a two-dimensional finite-element method. The key objective of this device is to identify slight differences in the refractive index by observing shifts in resonant wavelength through the light’s interaction with the plasmonic structure. Moreover, optimizing the geometric parameters significantly enhances the sensor’s performance. The optimized sensor achieves a maximum refractive index sensitivity of 2400 nm/RIU, a figure of merit (FOM) of 45.28, and a Q-factor of 39.86, as demonstrated by simulation results. Further investigations reveal the MIM structure’s capacity to detect adulteration in edible oils, underscoring the sensor’s adaptability and its potential for widespread biosensing applications.

Nanoscale Metal-Organic Frameworks as a Photoluminescent Platform for Bioimaging and Biosensing Applications.

The tracking of nanomedicines in their concentration and location inside living systems has a pivotal effect on the understanding of the biological processes, early-stage diagnosis, and therapeutic monitoring of diseases. Nanoscale metal-organic frameworks (nano MOFs) possess high surface areas, definite structure, regulated optical properties, rich functionalized sites, and good biocompatibility that allow them to excel in a wide range of biomedical applications. Controllable syntheses and functionalization endow nano MOFs with better properties as imaging agents and sensing units for the diagnosis and treatment of diseases. This minireview summarizes the tunable synthesis strategies of nano MOFs with controllable size, shape, and regulated luminescent performance, and pinpoints their recent advanced applications as optical elements in bioimaging and biosensing. The current limitations and future development directions of nano MOF-contained materials in bioimaging and biosensing applications are also discussed, aiming to expand the biological applications of nano MOF-based nanomedicine and facilitate their production or clinical translation.

Using Biosensors to Detect and Map Language Areas in the Brain for Individuals with Traumatic Brain Injury.

The application of biosensors in neurolinguistics has significantly advanced the detection and mapping of language areas in the brain, particularly for individuals with brain trauma. This study explores the role of biosensors in this domain and proposes a conceptual model to guide their use in research and clinical practice. The researchers explored the integration of biosensors in language and brain function studies, identified trends in research, and developed a conceptual model based on cluster and thematic analyses. Using a mixed-methods approach, we conducted cluster and thematic analyses on data curated from Web of Science, Scopus, and SciSpace, encompassing 392 articles. This dual analysis facilitated the identification of research trends and thematic insights within the field. The cluster analysis highlighted Functional Magnetic Resonance Imaging (fMRI) dominance and the importance of neuroplasticity in language recovery. Biosensors such as the Magnes 2500 watt-hour (WH) neuromagnetometer and microwire-based sensors are reliable for real-time monitoring, despite methodological challenges. The proposed model synthesizes these findings, emphasizing biosensors' potential in preoperative assessments and therapeutic customization. Biosensors are vital for non-invasive, precise mapping of language areas, with fMRI and repetitive Transcranial Magnetic Stimulation (rTMS) playing pivotal roles. The conceptual model serves as a strategic framework for employing biosensors and improving neurolinguistic interventions. This research may enhance surgical planning, optimize recovery therapies, and encourage technological advancements in biosensor precision and application protocols.

Molecular Mechanisms and Enhancement of Piezoelectricity in the M13 Virus

AbstractUnderstanding the structure and function of bioelectric materials is challenging due to the complex nature of biomaterials and a lack of appropriate tools. The precisely defined structures and genetic tunability of viruses provide an excellent model system to investigate bioelectrical behavior in biomaterials. This study presents the molecular mechanisms of piezoelectricity in the M13 bacteriophage (phage) under various mechanical stresses for bio‐piezoelectric generation. A computational approach is used to calculate the piezoelectric tensors of the M13 phage and quantify its direction‐dependent dipole moments. By computationally designing negatively charged residues on the phage surface, the surface charge density is enhanced to 16.7 µC cm−2. Using genetic engineering, phages are experimentally designed with different charges and tail structures to create model phage nanostructures, including individual phages, vertically standing phage films, and horizontally aligned phage films. Their vertical, horizontal, and shear‐mode piezoelectric properties are then measured using scanning probe microscopy techniques. The resulting phage‐based piezoelectric energy generators exhibit an effective piezoelectric coefficient of 15.4 pm V−1 and a power density of 4.2 µW cm−2. This phage‐based bioengineering approach provides a versatile platform for investigating fundamental mechanisms of bioelectricity and designing bioelectric materials for applications in energy harvesting, biomemory, and biosensors.

Microfluidic bubble-templating 3D printing of ordered macroporous hydrogels

Macroporous hydrogels have broad applications in tissue engineering, drug delivery and flexible biosensors, etc. However, it is still difficult to simultaneously control both the shape and the internal macroporosity. Here, we introduce a microfluidic bubble-templating 3D printing method based on thermosensitive composite hydrogel inks consisting of alginate and Pluronic F127. Two-phase laminar shear in coaxial microfluidics generates exceptionally monodispersed microbubbles as templates, while 3D printing technique provides spatial distribution of these microbubbles. Microbubble generation and 3D printing are coordinated and rheology of the inks are optimized to improve the systematic printability. Macroporous hydrogels with both monodispersed and gradient macropore structures are prepared and characterized. The internal anisotropic macroporosity distribution leads to inhomogeneous mechanical property in the products, making them competent as ergonomic artificial knuckle skins. The microfluidic-assisted 3D printing methodology provides a new insight for bottom-up design of porous soft materials.

Advances of Fluorescent Nanodiamond Platforms for Intracellular and On-Chip Biosensing.

Intracellular and extracellular sensing of physical and chemical variables is important for disease diagnosis and the understanding of cellular biology. Optical sensing utilizing fluorescent nanodiamonds (FNDs) is promising for probing intracellular and extracellular variables owing to their biocompatibility, photostability, and sensitivity to physicochemical quantities. Based on the potential of FNDs, we outlined the optical properties, biocompatibility, surface chemistry of FNDs and their applications in intracellular biosensing. This review also introduces biosensing platforms that combine FNDs and lab-on-a-chip approaches to control the extracellular environment and improve sample/reagent handling and sensing performance.