Search Results (716)

The phytohormone abscisic acid (ABA) plays a central role in organizing adaptive responses in plants to various abiotic stresses, helping the plant minimize the negative impact on growth and development. Rapid and direct detection of ABA is valuable for investigating plant responses to abiotic stress. In this work, we propose a novel label-free, non-competitive immunoassay for detecting and quantifying ABA easily and rapidly using a surface plasmon resonance (SPR) biosensor. The SPR sensor chip was functionalized with a commercial anti-ABA antibody, characterized for its affinity, binding kinetics, and specificity using the same platform. The direct assay demonstrated high specificity and sensitivity, with a calculated limit of detection of 1.36 ng/mL in buffer. The new immunosensor was applied to determine ABA concentrations directly in xylem sap samples from tomato plants subjected to abiotic stress (drought and high salinity) and was able to accurately reflect ABA levels corresponding to the applied stress. The results were comparable to the reference method, ultra-performance liquid chromatography coupled with tandem mass spectrometry (UPLC-MS/MS), establishing this new immunosensor as a novel detection method for rapid and reliable monitoring of ABA levels associated with abiotic stress in tomato plants. Full article

Acute Myocardial Infarction (AMI) is a critical cardiac condition that poses a substantial threat to myocardial function. Expedient diagnosis of AMI is paramount and relies on serological assays for rapid and accurate quantification of relevant biomarkers. Electrochemical sensors have emerged as promising candidates for this application, owing to their accessibility, operational simplicity, and high specificity. In this study, we developed a paper-based electrochemical immunosensor to detect cardiac troponin I in serum and saliva specimens. The electrode was fabricated using screen-printing technology with photographic paper as the substrate, employing graphite-based ink, nail polish, and acetone as the solvent. A quasi-reference electrode was constructed using silver powder-based ink, nail polish, and acetone. The immunosensor was prepared by modifying the working electrode with gold nanoparticles (AuNP) functionalized with cardiac troponin I antibodies (anti-cTnI) and bovine serum albumin (BSA). This modified electrode was subsequently used to detect the troponin I antigen. The analyses were performed in 0.1 mol L⁻¹ phosphate buffer medium, pH 7.00, in the presence of 5.0 mmol L⁻¹ of the potassium ferrocyanide probe. The immunosensor exhibited a sensitivity of 0.006 µA/fg mL⁻¹, a limit of detection of 9.83 fg mL⁻¹, and a limit of quantification of 32.79 fg mL⁻¹. Specificity studies conducted in the presence of other macromolecules demonstrated minimal interference, with relative standard deviations (RSD) below 5.00%, indicating a specific interaction with troponin I. Furthermore, the immunosensor demonstrated excellent reproducibility and stability. Upon application to serum and saliva samples, the immunosensor presented recoveries of approximately 99–105%, suggesting its potential applicability in clinical analyses. Full article

In this study, a colorimetric–photothermal dual-mode immunosensor based on Fe(Ⅲ)–gallic acid composite Prussian blue nanozyme (FGAN@PB NPs) was developed for the highly sensitive detection of malachite green (MG) in aquatic products. This strategy addresses the stability limitations associated with conventional horseradish peroxidase (HRP). In the colorimetric mode, the immunosensor exhibited an IC₅₀ of 7.56 ng/mL with a linear detection range of 2.21–25.84 ng/mL. In the photothermal mode, the linear range was 0.262–25.6 ng/mL, with a detection limit (LOD) of 0.31 ng/mL. The results from the two detection modes were mutually corroborative. Moreover, the detection of the proposed immunosensor was strongly correlated with the LC-MS/MS, offering a promising approach for the rapid on-site screening of MG and improving its applicability in complex sample matrices. Full article

Herein, we propose an ultrasensitive electrochemical immunosensor based on glucose oxidase labeling and enzyme-catalyzed Au staining. In brief, the primary antibody (Ab₁), bovine serum albumin, an antigen and then a bionanocomposite that contains a second antibody (Ab₂), poly(3-anilineboronic acid) (PABA), Au nanoparticles (AuNPs) and glucose oxidase (GOx) are modified on a glassy carbon electrode coated with multiwalled carbon nanotubes, yielding a corresponding sandwich-type immunoelectrode. In the presence of glucose, a chemical reduction of NaAuCl₄ by enzymatically generated H₂O₂ can precipitate a lot of gold on the Ab₂-PABA-AuNPs-GOx immobilized immunoelectrode. In situ anodic stripping voltammetry (ASV) detection of gold in 8 μL 1.0 M aqueous HBr-Br₂ is conducted for the antigen assay, and the ASV detection process takes approximately 6 min. This method is employed for the assay of human immunoglobulin G (IgG) and human carbohydrate antigen 125 (CA125), which demonstrates exceptional sensitivity, high selectivity and fewer required reagents/samples. The achieved limits of detection (S/N = 3) by the method are 0.25 fg mL⁻¹ for IgG (approximately equivalent to containing 1 IgG molecule in the 1 microlitre of the analytical solution) and 0.1 nU mL⁻¹ for CA125, which outperforms many previously reported results. Full article

Aflatoxin (AF) contamination in pistachios remains a critical food safety and trade challenge, given the potent carcinogenicity of AF-B₁ and the nut’s high susceptibility to Aspergillus infection throughout production and storage. Traditional decontamination methods such as roasting, irradiation, ozonation, and acid/alkaline treatments can reduce AF levels but often degrade sensory and nutritional quality, implying the need for more sustainable approaches. In recent years, innovative nonthermal interventions, including pulsed light, cold plasma, nanomaterial-based adsorbents, and bioactive coatings, have demonstrated significant potential to decrease fungal growth and AF accumulation while preserving product quality. Biosensing technologies such as electrochemical immunosensors, aptamer-based systems, and optical or imaging tools are advancing rapid, portable, and sensitive detection capabilities. Combining these experimental strategies with artificial intelligence (AI) and machine learning (ML) models can increasingly be applied to integrate spectral, sensor, and imaging data for predicting fungal development and AF risk in real time. This review brings together progress in nonthermal reduction strategies, biosensing innovations, and data-driven approaches, presenting a comprehensive perspective on emerging tools that could transform pistachio safety management and strengthen compliance with global regulatory standards. Full article

Monitoring contamination in soil and food systems remains vital for ensuring environmental and public health, particularly in agriculture-intensive regions. Existing laboratory-based techniques are often time-consuming, equipment-dependent, and impractical for rapid on-site screening. In this study, we present a portable, non-faradaic electrochemical impedance-based sensing platform capable of simultaneously detecting Salmonella Typhimurium (S. Typhi) and Aflatoxin B1 in spiked soil run-off samples. The system employs ZnO-coated electrodes functionalized with crosslinker for covalent antibody immobilization, facilitating selective, label-free detection using just 5 µL of sample. The platform achieves a detection limit of 1 CFU/mL for S. Typhi over a linear range of 10–10⁵ CFU/mL and 0.001 ng/mL for Aflatoxin B1 across a dynamic range of 0.01–40.96 ng/mL. Impedance measurements captured with a handheld potentiostat were strongly correlated with benchtop results (R² > 0.95), validating its reliability in field settings. The duplex sensor demonstrates high precision with recovery rates above 80% and coefficient of variation below 15% in spiked samples. Furthermore, machine learning classification of safe versus contaminated samples yielded an ROC-AUC > 0.8, enhancing its decision-making capability. This duplex sensing platform offers a robust, user-friendly solution for real-time environmental and food safety surveillance. Full article

Foodborne illnesses remain a global challenge, requiring rapid and sensitive detection platforms. We developed a magnetosome-based electrochemical immunosensor for lipopolysaccharide (LPS) antigens from Escherichia coli and Salmonella typhimurium. Magnetosomes isolated from Magnetospirillum sp. RJS1 were characterized by HR-TEM and functionalized with antibodies (2 CFU mL⁻¹), with FTIR confirming successful conjugation. The antibody–magnetosome complexes were immobilized on a chitosan/glutaraldehyde-modified glassy carbon electrode. AFM revealed globular (200–700 nm) and island-like (1–3 µm) features after antigen binding. Electrochemical impedance spectroscopy showed stepwise increases in charge-transfer resistance upon electrode modification and antigen interaction. The sensor exhibited high sensitivity toward E. coli (3–7 CFU mL⁻¹) and Salmonella (3–8 CFU mL⁻¹), achieving an immune sensitivity of 36.24 Ω/CFU mL⁻¹ and a detection limit of 1 CFU mL⁻¹. These results demonstrate the potential of magnetosome-based immunosensors as portable, efficient platforms for the rapid detection of foodborne pathogens in real samples. Full article

Carbendazim is a benzimidazole fungicide widely used in the prevention and control of vegetable diseases. However, if misused, it may result in residues in agricultural products, not only reducing vegetable quality but also posing potential risks to human health. Currently, the on-site rapid detection technology for carbendazim still faces challenges, including insufficient antibody specificity and low sensitivity, which hinder its ability to meet practical regulatory requirements. Therefore, this study screened a rational hapten structure by applying a computer-aided hapten design and obtained a specific antibody. Compared to previous studies, the cross-reactivity rate of the antibody with thiabendazole-methyl was less than 0.1%, and the cross-reactivity rate with 2-aminobenzimidazole was 52.7% lower than that of the existing reported antibodies, which significantly improved the detection specificity of the method. Based on a high-specificity antibody, a gold nanoparticle-based lateral flow immunoassay (AuNPs-LFIA) for carbendazim was established. The detection limits of green beans and leeks are 3.80 μg/kg and 1.80 μg/kg, respectively, which still maintain high specificity in complex samples. Good agreement was also demonstrated between the results of blind samples detected by AuNPs-LFIA and LC-MS/MS, respectively. The establishment of AuNPs-LFIA provides an effective solution for the rapid and specific detection of carbendazim. Full article

The real-time, cost-effective detection of marine toxins like tetrodotoxin (TTX) remains a significant challenge for the scientific community. Traditional methods, including cell-based assays (CBAs), high-performance liquid chromatography (HPLC), and automated patch clamp (APC), are time-consuming, requiring expensive lab-based equipment and highly trained personnel. Enzyme-linked immunosorbent assays (ELISAs), lateral flow assays (LFAs), and immunosensors may not be suitable for toxin analogues. Thus, a simplified approach has been developed in this study, which involves the electrophysiological and electrochemical interrogation of N2a cells grown on ITO-coated glass electrodes by measuring extracellular field potentials (EFP) in conjunction with whole-cell patch clamp recordings and electrochemical impedance spectroscopy (EIS) measurements both before and after incubation with TTX. The ITO substrate proved biocompatible and non-toxic for N2a cells. TTX exposure caused 102% inhibition in EFP values at 300 nM, confirmed by sodium current inhibition of 93% at 300 nM and 22% at 1 nM in patch clamp studies (IC₅₀ = 6.7 nM). EIS measurements indicated concentration-dependent impedance changes in the range of 6–300 nM. This research aims to provide a proof-of-concept for integration of electrophysiological and electrochemical approaches to simplify toxin detection systems. Full article

Lateral flow immunoassays (LFAs) are widely recognized as a powerful and versatile analytical platform. Nevertheless, the development of multiplex formats remains a distinct challenge. The aim of this study was to develop a multiplex LFA using gold nanoparticles (GNPs) as a label, selected for their ease of synthesis and functionalization with biomolecules. We provide practical recommendations regarding protein–hapten synthesis, membrane selection, application buffer composition, and methods to improve the long-term stability of the freeze-dried gold conjugate. The developed assay shows good tolerance to high-fat milk, stability at elevated temperatures, and promising sensitivity, with visual detection limits of 4–100 ng/mL for $\beta$-lactams, 1–10 ng/mL for tetracyclines, 50 ng/mL for streptomycin, and 0.3 ng/mL for chloramphenicol. Full article

Carbyne-containing materials offer significant potential for biosensor applications due to their unique chemical and mechanical properties. In this study, carbyne-enriched carbon coatings deposited on SiO₂/Si chips using ion-assisted pulse-plasma deposition were evaluated for the first time as substrates for optical biosensing. At first, the carbyne-enriched coatings were characterized by X-ray photoelectron spectroscopy, Raman spectroscopy, Atomic Force Microscopy, and the sessile drop method to assess their composition, structure, and wettability. After that, chips with carbyne-enriched coatings were modified with biomolecules through physical absorption or covalent bonding, and the respective biomolecular interactions were monitored in real-time by White Light Reflectance Spectroscopy (WLRS). In both cases, SiO₂/Si chips modified with an aminosilane were used as reference substrates. Physical adsorption was tested through immobilization of an antibody against C-reactive protein (CRP) to enable its immunochemical detection, whereas covalent bonding was tested through coupling of biotin and monitoring its reaction with streptavidin. It was found that the carbyne-enriched carbon-coated chips retained both their antibody adsorption capability and their covalent bonding ability for over 18 months, while the modified with aminosilane SiO₂/Si chips lost 90% of their antibody adsorption capacity and covalent bonding ability after two months of storage. These findings highlight the strong potential of carbyne-enriched carbon-coated chips as robust biosensing substrates, with applications extending beyond WLRS. Full article

We report herein the development of a polyclonal antibody against ochratoxin A (OTA) using a specially designed synthetic OTA derivative as the immunizing hapten. This OTA derivative contains a tetrapeptide linker (glycyl-glycyl-glycyl-lysine, GGGK), through which it can be linked to a carrier protein and form an immunogenic conjugate. The OTA derivative (OTA-glycyl-glycyl-glycyl-lysine, OTA-GGGK) has been synthesized on a commercially available resin via the well-established Fmoc-based solid-phase peptide synthesis (Fmoc-SPPS) strategy; overall, this approach has allowed us to avoid tedious liquid-phase synthesis protocols, which are often characterized by multiple steps, several intermediate products and low overall yield. Subsequently, OTA-GGGK was conjugated to bovine thyroglobulin through glutaraldehyde, and the conjugate was used in an immunization protocol. The antiserum obtained was evaluated with a simple-format ELISA in terms of its titer and capability of recognizing the natural free hapten; the anti-OTA antibody, as a whole IgG fragment, was successfully applied to three different immunoanalytical systems for determining OTA in various food materials and wine samples, i.e., a multi-mycotoxin microarray bio-platform, an optical immunosensor, and a biotin–streptavidin ELISA, which has proved the analytical effectiveness and versatility of the anti-OTA antibody developed. The same approach may be followed for developing antibodies against other low-molecular-weight toxins and hazardous substances. Full article

A competitive electrochemical immunosensor, using screen-printed carbon electrodes (SPCEs), was developed for the determination of total fumonisins (sum of FB1, FB2 and FB3) extracted with a simple solvent extraction and dilution method, without clean up, from maize flour and maize tortillas. The optimized biosensor has a linear range of 0.25 to 50 µg/L with 3% and 2% reproducibility for FB1 and (FB1 + FB2), respectively, and a linear range of 0.25 to 10 µg/L with 2% reproducibility for (FB1 + FB2 + FB3). The limits of detection and quantification in PBS buffer for total fumonisins are 0.12 µg/L and 0.39 µg/L, respectively. These values in the maize matrix are 6.07 µg/kg and 20.25 µg/kg, respectively. In addition, the stability and the selectivity of the sensor were studied. The immunosensor was validated with liquid chromatography–tandem mass spectrometry. This novel biosensor is more rapid, simpler and cheaper than current methods, and can also be used at the point of need. Full article

Since carbohydrate antigen 19-9 (CA 19-9) is a significant biomarker for the clinical diagnosis and treatment of pancreatic cancer, a sensitive sandwich-type immunosensor was proposed with an epoxy functionalized covalent organic framework (EP-COF_TTA-DHTA) as the antibody carrier and an electroactive COF_{TTA-2,6-NA(OH)2} as the signal amplification probe for the sensitive detection of CA 19-9. The flexible covalent linkage between the epoxy-functionalized EP-COF_TTA-DHTA and the antibodies was employed to improve the dynamics of the antigen–antibody interaction significantly. Meanwhile, AuNPs@COF_{TTA-2,6-NA(OH)2} with abundant electroactive sites enhanced the current response of the immunoreaction significantly. After optimizing the incubation time and concentration of the antibody, CA 19-9 was quantitatively detected by differential pulse voltammetry (DPV) based on the sensitive sandwich-type immunosensor with a low detection limit of 0.0003 U/mL and a wide linear range of 0.0009–100 U/mL. The electrochemical immunosensor exhibits high specificity, stability and repeatability, and it provides a feasible and efficient method for the pathologic analysis and treatment of tumor markers. Full article