Search Results (59)

Protein biomarkers can be used for monitoring the occurrence and development of diseases. Accurate, sensitive, and low-cost methods for protein detection can facilitate therapeutic intervention, improve clinical outcome, and reduce economic pressure for patients. Molecularly imprinted polymers (MIPs) have been considered as a type of biomimetic materials for developing biosensing technologies due to their advantages of high stability, low preparation cost, and good reusability over classical biometric recognition elements such as antibodies and aptamers. Electrochemical biosensors have become the most promising technology in sensing applications in view of their high sensitivity, fast response speed, cost-effectiveness, good stability, and ease of miniaturization. Efforts have been made to develop various electrochemical biosensors for protein detection with MIPs as recognition elements. This article provides an overview of the progress in molecular imprinting methods for the design and application of electrochemical protein biosensors. The strategies for imprinting and removing templates and preparing MIPs-modified sensing electrodes are comprehensively discussed. Finally, the challenges and future perspectives of protein-imprinted electrodes are addressed. This work will contribute to the development of innovative analytical devices based on MIPs for monitoring and managing various diseases by determining protein biomarkers. Full article

This study uses a bismuth ferrite (BiFeO₃) and MXene (Ti₃C₂Tx) to design a surface plasmon resonance (SPR) biosensor for the sensitivity enhancement at a 633 nm wavelength. Here, MXene serves as a biorecognition element (BRE) layer to ensure stable and reliable biomolecule adsorption. The MXene is a family of two-dimensional (2D) materials with metallic-like conductivity, a large surface area that can attach biomolecules, and improve biocompatibility. The addition of a conductive 2D MXene layer and a high-index BiFeO₃ dielectric layer greatly improves light–matter interaction and evanescent field penetration at the sensing interface. Strong plasmonic coupling is indicated by the reflectance analysis, which shows a distinct and consistent shift in the resonance angle as analyte RI increases. This study examined the sensitivity at optimized Ag and BiFeO₃ layer thickness. At an Ag of 39 nm and BiFeO₃ of 3 nm thickness, the maximal sensitivity of 340.68°/RIU with a remarkable figure of merit (FoM) of 47.38/RIU is obtained. The overall detection accuracy (DA) and FoM are significantly improved by the large sensitivity enhancement, despite a slight increase in full width at half maximum (FWHM). Furthermore, the penetration depth (PD) of 198.50 nm (at RI:1.330) and 199.52 nm (at RI:1.335) is attained with the proposed structure. Due to its high sensitivity, reusability, and reproducibility, the SPR biosensor has the potential to be used in biochemical, environmental, and medical detection. Full article

The rapid and accurate detection of SARS-CoV-2 biomarkers remains a critical requirement for effective outbreak control and decentralized diagnostics. Although RT-PCR is the current gold standard, its reliance on centralized laboratories and long processing times limits its applicability in point-of-care settings. In this context, optical biosensing platforms based on surface plasmon resonance (SPR) offer attractive features, including label-free, real-time, and quantitative detection. This study explores the use of synthetic receptors for the highly sensitive detection of the receptor-binding domain (RBD) of the SARS-CoV-2 spike protein. Specifically, soft molecularly imprinted polymer nanoparticles (nanoMIPs) were employed as synthetic receptors and integrated into a high-sensitivity, portable plasmonic platform based on a D-shaped plastic optical fiber (POF) SPR sensor. The nanoMIPs were selectively imprinted against the RBD, characterized by Dynamic Light Scattering (DLS), Isothermal Titration Calorimetry (ITC), and Scanning Electron Microscopy (SEM) to confirm nanoMIPs size, binding properties, and surface morphology. Next, the nanoMIPs were immobilized onto a gold-coated sensing surface, enabling enhanced specificity, affinity, and signal amplification compared to conventional biological recognition elements. The resulting RBD-SPR-nanoMIPs sensor demonstrated promising analytical performance, exhibiting high selectivity against potentially interfering proteins and an anticipated sensitivity suitable for RBD detection at femtomolar concentrations. The inherent stability of nanoMIPs suggests the potential for reusable SPR sensing platforms, paving the way for next-generation synthetic receptor-based plasmonic biosensors. Full article

The reusability of enzymes is a fundamental aspect of sustainable biotechnology and the development of biosensors. This study presents one of the first quantitative evaluations of DNA polymerase reusability by utilizing integrated quartz crystal microbalance (QCM) kinetics and real-time monitoring of exonuclease activity. The results showed that immobilized T7 DNA polymerase retained approximately 50% of its initial activity after three 90-min cycles and around 20% after five cycles. Significantly lower activities were observed for shorter, 45-min cycles. This indicates an unexpected time-dependent enhancement in stability for longer reaction times. The findings suggest a promising trend in enzyme stability and reusability, establishing a quantitative relationship between reaction duration and enzyme performance. This relationship offers a scalable pathway for the regeneration of biosensors and for sustainable enzymatic catalysis. Additionally, the work provides a transferable framework that can be applied to other DNA-processing enzymes, which supports long-term biosensor performance and industrial biocatalysis. The demonstrated approach offers a transferable and scalable methodology for the development of reusable polymerase-based biosensors and sustainable biocatalytic systems. Full article

Over the past few decades, Fc fragment-conjugated proteins, such as Protein A, have been extensively utilized across a range of applications, including antibody purification, site-specific immobilization of antibodies, and the development of biosensing platforms. In this study, building upon our group prior research, we designed and screened an affinity DNA functional ligand (A-DNAFL) and experimentally validated its binding affinity (K_D = 6.59 × 10⁻⁸) toward mouse IgG antibodies, whose binding performance was comparable to that of protein A. Systematic evaluations were performed to assess the binding efficiency under varying pH levels and ionic strength conditions. Optimal antibody immobilization was achieved in PBST-B buffer under physiological pH 7.2–7.4 and containing approximately 154 mM Na⁺ and 4 mM K⁺. Two competitive binding assays confirmed that the A-DNAFL binds to the Fc fragment of murine IgG antibody. Furthermore, molecular docking simulations were employed to investigate the interaction mode, revealing key residues involved in binding as well as the contributions of hydrogen bonding and hydrophobic interactions to complex stabilization. Leveraging these insights, A-DNAFL was utilized as a tool for oriented immobilization of antibodies on the sensing interface, enabling the construction of an immunosensor for ricin detection. Following optimization of immobilization parameters, the biosensor exhibited a detection limit of 30.5 ng/mL with the linear regression equation is lg(Response) = 0.329 lg(C_ricin) − 2.027 (N = 9, R = 0.938, p < 0.001)—representing a 64-fold improvement compared to conventional protein A-based methods. The system demonstrated robust resistance to nonspecific interference. Sensing interface reusability was also evaluated, showing only 8.55% signal reduction after two regeneration cycles, indicating that glycine effectively elutes bound antibodies while preserving sensor activity. In summary, the A-DNAFL presented in this study represents a novel antibody-directed immobilization material that serves as a promising alternative to protein A. It offers several advantages, including high modifiability, low production cost, and a relatively small molecular weight. These features collectively contribute to its broad application potential in biosensing, antibody purification, and other areas of life science research. Full article

Enzyme-based biosensors have emerged as a transformative technology, leveraging the specificity and catalytic efficiency of enzymes across various domains, including medical diagnostics, environmental monitoring, food safety, and industrial processes. These biosensors integrate biological recognition elements with advanced transduction mechanisms to provide highly sensitive, selective, and portable solutions for real-time analysis. This review explores the key components, detection mechanisms, applications, and future trends in enzyme-based biosensors. Artificial enzymes, such as nanozymes, play a crucial role in enhancing enzyme-based biosensors by mimicking natural enzyme activity while offering improved stability, cost-effectiveness, and scalability. Their integration can significantly boost sensor performance by increasing the catalytic efficiency and durability. Additionally, lab-on-a-chip and microfluidic devices enable the miniaturization of biosensors, allowing for the development of compact, portable devices that require minimal sample volumes for complex diagnostic tests. The functionality of enzyme-based biosensors is built on three essential components: enzymes as biocatalysts, transducers, and immobilization techniques. Enzymes serve as the biological recognition elements, catalyzing specific reactions with target molecules to produce detectable signals. Transducers, including electrochemical, optical, thermal, and mass-sensitive types, convert these biochemical reactions into measurable outputs. Effective immobilization strategies, such as physical adsorption, covalent bonding, and entrapment, enhance the enzyme stability and reusability, enabling consistent performance. In medical diagnostics, they are widely used for glucose monitoring, cholesterol detection, and biomarker identification. Environmental monitoring benefits from these biosensors by detecting pollutants like pesticides, heavy metals, and nerve agents. The food industry employs them for quality control and contamination monitoring. Their advantages include high sensitivity, rapid response times, cost-effectiveness, and adaptability to field applications. Enzyme-based biosensors face challenges such as enzyme instability, interference from biological matrices, and limited operational lifespans. Addressing these issues involves innovations like the use of synthetic enzymes, advanced immobilization techniques, and the integration of nanomaterials, such as graphene and carbon nanotubes. These advancements enhance the enzyme stability, improve sensitivity, and reduce detection limits, making the technology more robust and scalable. Full article

Polyphenol oxidase (PPO) is a metalloproteinase widely present in plant organelles that plays crucial roles in photosynthesis, pest and disease resistance, growth and development, and flower color formation. Due to the high cost and reuse difficulties of plant PPO in applications, immobilization has emerged as a key technology to improve its stability, recyclability, and reusability. Immobilized plant PPO has been widely used in environmental and detection fields. This review examines different immobilization methods and carrier materials for plant PPO and summarizes its applications in wastewater treatment, biosensor detection, food preservation, and theaflavin synthesis. Finally, current challenges and future opportunities for immobilized plant PPO are discussed. Full article

Metal–organic framework (MOF)-based nanozymes represent a groundbreaking frontier in advanced microbial biosensing, offering unparalleled catalytic precision and structural tunability to mimic natural enzymes with superior stability and specificity. By engineering the structural features and forming composites, MOFs are precisely tailored to amplify nanozymatic activity, enabling the highly sensitive, rapid, and cost-effective detection of a broad spectrum of microbial pathogens critical to biomedical diagnostics and environmental monitoring. These advanced biosensors surpass traditional enzyme systems in robustness and reusability, integrating seamlessly with smart diagnostic platforms for real-time, on-site microbial identification. This review highlights cutting-edge developments in MOF nanozyme design, composite engineering, and signal transduction integration while addressing pivotal challenges such as biocompatibility, complex matrix interference, and scalable manufacturing. Looking ahead, the convergence of multifunctional MOF nanozymes with portable technologies and optimized in vivo performance will drive transformative breakthroughs in early disease detection, antimicrobial resistance surveillance, and environmental pathogen control, establishing a new paradigm in next-generation smart biosensing. Full article

Breast cancer is a leading cause of cancer-related mortality worldwide, requiring efficient diagnostic tools for early detection and monitoring. Human epidermal growth factor receptor 2 (HER2) is a key biomarker for breast cancer classification, typically assessed using immunohistochemistry (IHC). However, IHC requires invasive biopsies and time-intensive laboratory procedures. In this study, we present a biosensor integrated with a reusable printed circuit board (PCB) and functionalized glucose test strips designed for rapid and non-invasive HER2 detection in saliva. The biosensor achieved a limit of detection of 10⁻¹⁵ g/mL, 4 to 5 orders of magnitude more sensitive than the enzyme-linked immunosorbent assay (ELISA), with a sensitivity of 95/dec and a response time of 1 s. In addition to HER2, the biosensor also detects cancer antigen 15-3 (CA15-3), another clinically relevant breast cancer biomarker. The CA15-3 test demonstrated an equally low limit of detection, 10⁻¹⁵ g/mL, and a higher sensitivity, 190/dec, further validated using human saliva samples. Clinical validation using 29 saliva samples confirmed our biosensor’s ability to distinguish between healthy, in situ breast cancer, and invasive breast cancer patients. The system, which integrates a Bluetooth Low-Energy (BLE) module, enables remote monitoring, reduces hospital visits, and enhances accessibility for point-of-care and mobile screening applications. This ultra-sensitive, rapid, and portable biosensor can serve as a promising alternative for breast cancer detection and monitoring, particularly in rural and underserved communities. Full article

Polymer coatings as thin films stand out as a commonly used strategy to modify biosensor surfaces for improving detection performance; however, nonspecific biomolecule interactions and the limited degree of ligand conjugation on the surface have necessitated the development of innovative methods for surface modification. To this end, methacrylated tethered telechelic polyethylene glycol (PEG-diMA) chains of three different molecular weights (2, 6, and 10 kDa) were synthesized herein and used for obtaining thiolated nanoparticles (NPs) upon adding excess amounts of a tetra-thiol crosslinker. Characterized according to their size, surface charge, morphology, and thiol amounts, these nanoparticles were immobilized on gold surfaces that mimicked gold-coated mass sensor platforms. The PEG-based nanoparticles, prepared especially by PEG6K-diMA polymers, were shown to result in the preparation of a monolayer and smooth coating of 80–120 nm thickness. Cysteine-modified NTS(8–13) peptide (RRPYIL) was conjugated to thiolated NP with reversible disulfide bonds and it was demonstrated that its cleavage with a reducing agent such as dithiothreitol (DTT) restores the NP-immobilized gold surface for at least two cycles. Together with its binding studies to NTSR2 antibodies, it was revealed that the peptide-conjugated NP-modified gold surface could be employed as a model for a reusable sensor surface for the detection of biomarkers of same or different types. Full article

Depression and anxiety are two common mental health issues that require serious attention, as they have significant impacts on human well-being, with both being emotionally and physically reflected in the increasing number of suicide cases globally. The World Health Organization (WHO) estimated that about 322 million people around the world experienced mental illnesses in 2017, and this number continues to increase. Cortisol is a major stress-controlled hormone that is regulated by the hypothalamic–pituitary–adrenal (HPA) axis. The HPA axis has three main components, including the hypothalamus, pituitary gland, and adrenal gland, where cortisol, the primary stress hormone, is released. It plays crucial roles in responding to stress, energy balance, and the immune system. The cortisol level in the bloodstream usually increases when stress develops. Molecularly imprinted polymers (MIPs) have been highlighted in terms of creating artificial bioreceptors by mimicking the shape of detected biomolecules, making natural bioreceptor molecules no longer required. MIPs can overcome the limitations of chemicals and physical properties reducing over time and the short-time shelf life of natural bioreceptors. MIPs’ benefits are reflected in their ease of use, high sensitivity, high specificity, reusability, durability, and the lack of requirement for complicated sample preparation before use. Moreover, MIPs incur low costs in manufacturing, giving them a favorable budget for the market with simple utilization. MIPs can be formulated by only three key steps, including formation, the polymerization of functional monomers, and the creation of three-dimensional cavities mimicking the shape and size of targeting molecules. MIPs have a high potential as biosensors, especially working as bioanalytics for protein, anti-body, antigen, or bacteria detection. Herein, this research proposes an MIP-based cortisol biosensor in which cortisol is imprinted on methyl methacrylate (MMA) and methacrylic acid (MAA) produced by UV polymerization. This MIP-based biosensor may be an alternative method with which to detect and monitor the levels of hormones in biological samples such as serum, saliva, or urine due to its rapid detection ability, which would be of benefit for diagnosing depression and anxiety and prescribing treatment. In this study, quantitative detection was performed using an electrochemical technique to measure the changes in electrical signals in different concentrations of a cortisol solution ranging from 0.1 to 1000 pg/mL. The MIP-based biosensor, as derived by calculation, achieved its best detection limit of 1.035 pg/mL with a gold electrode. Tests were also performed on molecules with a similar molecular structure, including Medroxyprogesterone acetate and drospirenone, to ensure the sensitivity and accuracy of the sensors, demonstrating a low sensitivity and low linear response. Full article

Tetracyclines (TCs) are antibiotics used extensively in medicine, veterinary science, and animal husbandry. Their overuse and the widespread presence of their residues in the environment contribute to intensifying the phenomenon of antibiotic resistance (ABR). The efforts are being made to reduce the spread of antibiotics and control the phenomenon of ABR, and one of the key methods is monitoring the presence of antibiotic residues in the environment and food of animal origin. Herein, we provide the overview of the recent developments in electrochemical (bio)sensing of tetracyclines in different types of samples. The review presents a comprehensive view of such aspects of the practical (bio)sensor application as sample preparation, the reusability of (bio)sensors, and the possibility of determining antibiotics at levels required by regulations. Advances, existing challenges, and future trends in the development of novel (bio)electrochemical methods of tetracycline quantification were discussed. Full article

Glycine is an essential amino acid involved in synthesizing a variety of important biomolecules, and its concentration can influence numerous biochemical processes, including the severity of symptoms in a wide range of conditions in humans, such as cancer, schizophrenia, major depression, and diabetes. While a few costly or labour-intensive methods are currently available, we have developed a new enzymatic biosensor that can accurately measure glycine levels with remarkable simplicity. By employing immobilized laccase enzymes in combination with a surface plasmon resonance (SPR) device, our system achieved a limit of detection (LOD) of 9.95 mM and a limit of quantification (LOQ) of 33.19 mM. In addition, it demonstrated a recovery rate of 97.64 ± 7.71%. Moreover, the biosensor maintained consistent signal intensity over 21 days and supported a total of 60 analyses using the same immobilized enzyme setup, demonstrating excellent reusability. Notably, this study marks the first time glycine has been determined using an enzymatic SPR-based platform. Full article

Biosensors are transforming point-of-care diagnostics by simplifying the detection process and enabling rapid, accurate testing. This study introduces a novel, reusable biosensor designed for direct viral RNA detection from unfiltered saliva, targeting SARS-CoV-2. Unlike conventional methods requiring filtration, our biosensor leverages a unique electrode design that prevents interference from saliva debris, allowing precise measurements. The biosensor is based on electrochemical principles, employing oligonucleotide probes immobilized on a hydrophobic-coated electrode, which prevents air bubbles and salt crystal formation. During validation, the biosensor demonstrated a sensitivity and specificity of 100%, accurately identifying SARS-CoV-2 in saliva samples without false positives or negatives. Cross-validation with RT-qPCR, the gold standard for COVID-19 diagnostics, confirmed the reliability of our device. The biosensor’s performance was tested on 60 participants, yielding 12 true positive results and 48 true negatives, aligning perfectly with RT-qPCR outcomes. This reusable, easy-to-use biosensor offers significant potential for point-of-care applications in various healthcare settings, providing a fast, efficient, and cost-effective method for detecting viral infections such as COVID-19. Its robust design, minimal sample preparation requirements, and multiple-use capability mark a significant advancement in biosensing technology. Full article

Vascular endothelial growth factor (VEGF) is a critical angiogenesis biomarker associated with various pathological conditions, including cancer. This study leverages pre-biotinylated FcγRI interactions with IgG1-type monoclonal antibodies to develop a sensitive VEGF detection method. Utilizing surface plasmon resonance (SPR) technology, we characterized the binding dynamics of immobilized biotinylated FcγRI to an IgG1-type antibody, Bevacizumab (AVT), through kinetic studies and investigated suitable conditions for sensor surface regeneration. Subsequently, we characterized the binding of FcγRI-captured AVT to VEGF, calculating kinetic constants and binding affinity. A calibration curve was established to analyze the VEGF quantification capacity and accuracy of the biosensor, computing the limits of blank, detection, and quantification at a 95% confidence interval. Additionally, the specificity of the biosensor for VEGF over other protein analytes was assessed. This innovative biomimetic approach enabled FcγRI-mediated site-specific AVT capture, establishing a stable and reusable platform for detecting and accurately quantifying VEGF. The results indicate the effectiveness of the plasmonic sensor platform for VEGF detection, making it suitable for research applications and, potentially, clinical diagnostics. Utilizing FcγRI-IgG1 antibody binding, this study highlights the industrial and clinical value of advanced biosensing technologies, offering insights to enhance therapeutic monitoring and improve outcomes in anti-VEGF therapies. Full article