Recent Advances in Development of Biosensors for Monitoring of Airborne Microorganisms.

Background: The early detection of infectious microorganisms is crucial for preventing and controlling the transmission of diseases. This article provides a comprehensive review of biosensors based on various diagnostic methods for measuring airborne pathogens. Objective: This article aims to explore recent advancements in the field of biosensors tailored for the detection and monitoring of airborne microorganisms, offering insights into emerging technologies and their potential applications in environmental surveillance and public health management. Materials and Methods: The study summarizes the research conducted on novel methods of detecting airborne microorganisms using different biological sensors, as well as the application of signal amplification technologies such as polymerase chain reaction (PCR), immunoassay reactions, molecular imprinted polymers (MIP) technique, lectin and cascade reactions, and nanomaterials. Results: Antibody and PCR detection methods are effective for specific microbial strains, but they have limitations including limited stability, high cost, and the need for skilled operators with basic knowledge of the target structure. Biosensors based on MIP and lectin offer a low-cost, stable, sensitive, and selective alternative to antibodies and PCR. However, challenges remain, such as the detection of small gas molecules by MIP and the lower sensitivity of lectins compared to antibodies. Additionally, achieving high sensitivity in complex environments poses difficulties for both methods. Conclusion: The development of sensitive, reliable, accessible, portable, and inexpensive biosensors holds great potential for clinical and environmental applications, including disease diagnosis, treatment monitoring, and point-of-care testing, offering a promising future in this field. This review presents an overview of biosensor detection principles, covering component identification, energy conversion principles, and signal amplification. Additionally, it summarizes the research and applications of biosensors in the detection of airborne microorganisms. The latest advancements and future trends in biosensor detection of airborne microorganisms are also analyzed.

Bioelectrochemical biosensors for water quality assessment and wastewater monitoring.

Bioelectrochemical biosensors offer a promising approach for real-time monitoring of industrial bioprocesses. Many bioelectrochemical biosensors do not require additional labelling reagents for target molecules. This simplifies the monitoring process, reduces costs, and minimizes potential contamination risks. Advancements in materials science and microfabrication technologies are paving the way for smaller, more portable bioelectrochemical biosensors. This opens doors for integration into existing bioprocessing equipment and facilitates on-site, real-time monitoring capabilities. Biosensors can be designed to detect specific heavy metals such as lead, mercury, or chromium in wastewater. Early detection allows for the implementation of appropriate removal techniques before they reach the environment. Despite these challenges, bioelectrochemical biosensors offer a significant leap forward in wastewater monitoring. As research continues to improve their robustness, selectivity, and cost-effectiveness, they have the potential to become a cornerstone of efficient and sustainable wastewater treatment practices.

An Integrated All-Natural Conductive Supramolecular Hydrogel Wearable Biosensor with Enhanced Biocompatibility and Antibacterial Properties.

Conductive hydrogels exhibit tremendous potential for wearable bioelectronics, biosensing, and health monitoring applications, yet concurrently enhancing their biocompatibility and antimicrobial properties remains a long-standing challenge. Herein, we report an all-natural conductive supramolecular hydrogel (GT5-DACD2-B) prepared via the Schiff base reaction between the biofriendly dialdehyde cyclodextrin and gelatin. The potent antibacterial agent fusidic acid (FA) is incorporated through host-guest inclusion, enabling 100% inhibition of Staphylococcus aureus proliferation. The biocompatibility of our hydrogel is bolstered with tannic acid (TA) facilitating antibacterial effects through interactions with gelatin, while borax augments conductivity. This supramolecular hydrogel not only exhibits stable conductivity and rapid response characteristics but also functions as a flexible sensor for monitoring human movement, facial expressions, and speech recognition. Innovatively integrating biocompatibility, antimicrobial activity, and conductivity into a single system, our work pioneers a paradigm for developing multifunctional biosensors with integrated antibacterial functionalities, paving the way for advanced wearable bioelectronics with enhanced safety and multifunctionality.

pH-responsive persistent luminescence nanoprobes biosensor for autofluorescence-free determination of glucose level in human samples and fingerprint encryption.

Glucose level is an important indicator of diabetes, and maintaining an appropriate physiological concentration of glucose is important for human health. However, traditional optical sensors are interfered by the interference of strong background autofluorescence and natural responsive luminescence, which severely limits their application in complex biological samples. Herein, as a novel glucose biosensing probe, green-emitting Zn2GeO4:Mn2+, Eu3+ (ZGME) persistent luminescence nanoparticles (PLNPs) with pH stimulus-responsive was prepared by a facile one-pot hydrothermal method. We also investigated the pH stimulus-responsive luminescence behaviour of ZGME over a range of pH values from 2.8 to 8.0. Taking advantage of the interesting property that ZGME photoluminescence intensity has a pH response, within an extraordinarily narrow pH range of 5.0-6.5 for highly selectivity and sensitive determination of glucose level in human samples by acid-responsive quenching and persistent luminescent performance. The detection results show high accuracy of the measured values of glucose in serum with a wide detection range (2.5 µg L-1-10 mg L-1) and low detection limit (0.5 µg L-1). Finally, the pH-responsive persistent luminescence also makes ZGME promising for high-level fingerprint information encryption. Hence, the established pH stimulation-responsive PLNPs-based biosensing probe offers excellent performance with high selective, accuracy and signal-to-noise ratio for detection of glucose level in human samples.

Optical Based Surface Plasmon Resonance Sensor for the Detection of the Various Kind of Cancerous Cell.

This research explores a novel biosensor design that exhibits much higher sensitivity compared to conventional biosensors. The biosensor's uniqueness originated from its innovative structure, which incorporates N-FK51A/Ag/AlON/BlueP materials, as well as its cutting-edge fabrication method. The refractive index component was considered when designing the SPR biosensor, which was developed from the angular analysis of the attenuated total reflection (ATR) approach for cancer detection. For instance, the resonance angle shifts by 15.57 deg when the refractive index changes from 1.360 to 1.401, demonstrating the sensor's responsiveness to variation in the refractive index. The sensitivities for skin (basal), cervical (HeLa), blood (Jurkat), adrenal gland (PC12), and breast (MDA-MB-231 and MCF-7) cancer cells were 197.65, 243.66, 255.36, 302.71, 372.57, and 416.85 deg/RIU, respectively. Also, the detection accuracy (DA), the figure of merit (FoM), and the quality factor were 0.37/deg, 155.94 (deg/RIU), and 26.71 RIU-1. We also examine the effects of substituting the noble, dielectric, 2D material layer with conventional biosensor materials for six cancers. Each time, the Ag/AION/BlueP layered structure performed best in distinguishing cancer cells from healthy cells. We also study the prism effects. The proposed biosensor, with a RI of 1.29-1.40, has a linear regression coefficient of R2 of 0.96094.

Disposable glutamate biosensor based on platinum nanoparticles, carbon quantum dots and poly-L-aspartic acid.

This study reports the design and development of a disposable amperometric biosensor for the determination of L-glutamate. Glutamate oxidase (GlOx) was immobilized onto a screen-printed carbon electrode (SPE) modified with poly-L-Aspartic acid (PAsp), carbon quantum dots (CQD), and platinum nanoparticles (PtNP) for the construction of the biosensor. The surface composition of the modified SPE was optimized using the one variable at a time method. The morphological properties of the biosensor were characterized by scanning electron microscopy and energy-dispersive X-ray spectroscopy. The electrochemical behavior of the modified electrodes was studied by cyclic voltammetry. Under the optimized experimental conditions the linear working range, detection limit and sensitivity of the GlOx/PtNP/CQD/PAsp/SPE were found to be 1.0 - 140 µM, 0.3 µM and 0.002 µA µM-1, respectively. The GlOx/PtNP/CQD/PAsp/SPE biosensor also exhibited good measurement repeatability. The as-developed biosensor was applied for the determination of L-glutamate in spiked serum samples and the average analytical recovery of added glutamate was 98.9 ± 3.9%.

Systematic Study of Various Functionalization Steps for Ultrasensitive Detection of SARS-CoV-2 with Direct Laser-Functionalized Au-LIG Electrochemical Sensors.

The 2019 coronavirus (COVID-19) pandemic impaired global health, disrupted society, and slowed the economy. Early detection of the infection using highly sensitive diagnostics is crucial in preventing the disease's spread. In this paper, we demonstrate electrochemical sensors based on laser induced graphene (LIG) functionalized directly with gold (Au) nanostructures for the detection of SARS-CoV-2 with an outstanding limit of detection (LOD) of ∼1.2 ag·mL-1. To achieve the optimum performance, we explored various functionalization parameters to elucidate their impact on the LOD, sensitivity, and linearity. Specifically, we investigated the effect of (i) gold precursor concentration, (ii) cross-linker chemistry, (iii) cross-linker and antibody incubation conditions, and (iv) antigen-sensor interaction (diffusion-dominated incubation vs pipette-mixing), as there is a lack of a systematic study of these parameters. Our benchmarking analysis highlights the critical role of the antigen-sensor interaction and cross-linker chemistry. We showed that pipette-mixing enhances sensitivity and LOD by more than 1.6- and 5.5-fold, respectively, and also enables multimodal readout compared to diffusion-dominated incubation. Moreover, the PBA/Sulfo-NHS: EDC cross-linker improves the sensitivity and LOD compared to PBASE. The sensors demonstrate excellent selectivity against other viruses, including HCoV-229E, HCoV-OC43, HCoV-NL63, and influenza H5N1. Beyond the ability to detect antigen fragments, our sensors enable the detection of antigen-coated virion mimics (which are a better representative of the real infection) down to an ultralow concentration of ∼5 particles·mL-1.

Enhanced enzymatic electrochemical detection of an organophosphate Pesticide: Achieving Wide linearity and femtomolar detection via gold nanoparticles growth within polypyrrole films.

The indiscriminate use of pesticides in agriculture demands the development of devices capable of monitoring contaminations in food supplies, in the environment and biological fluids. Simplicity, easy handling, high sensitivities, and low limits-of-detection (LOD) and quantification are some of the required properties for these devices. In this work, we evaluated the effect of incorporating gold nanoparticles into indigo carmine-doped polypyrrole during the electropolymerization of films for use as an acetylcholinesterase (AChE) enzyme-based biosensor. As proof of concept, the pesticide methyl parathion was tested towards the inhibition of AChE. The enzyme was immobilized simply by drop-casting a solution, eliminating the need for any prior surface modification. The biosensors were characterized with cyclic voltammetry, scanning electron microscopy, transmission electron microscopy, and Raman spectroscopy. The assays for the detection of methyl parathion with films containing polypyrrole, indigo carmine and AChE (PPy-IC-AChE) presented a sensitivity of 5.7 µA cm-2 g-1 mL and a LOD of 12 nmol L-1 (3.0 ng L-1) with a linear range from 1.3 x 10-7 mol L-1 to 1.0 x 10-5 mol L-1. The introduction of gold nanoparticles (AuNP) into the film (PPy-IC-AuNP-AChE) led to remarkable improvements on the overall performance, such as a lower redox potential for the enzymatic reaction, a 145 % increase in sensitivity (14 µA cm-2 g-1 mL), a wider detection dynamic range (from 1.3x10-7 to 1.0x10-3 mol L-1), and a very low LOD of 24 fmol L-1 (64 ag mL-1). These findings underscore the potential of using AuNPs to improve the enzymatic performance of biosensor devices.

Pumpless microfluidic sweat sensing yarn.

Textile sweat sensors possess immense potential for non-invasive health monitoring. Rapid in-situ sweat capture and prevention of its evaporation are crucial for accurate and stable real-time monitoring. Herein, we introduce a unidirectional, pump-free microfluidic sweat management system to tackle this challenge. A nanofiber sheath layer on micrometer-scale sensing filaments enables this pumpless microfluidic design. Utilizing the capillary effect of the nanofibers allows for the swift capture of sweat, while the differential configuration of the hydrophilic and hydrophobic properties of the sheath and core yarns prevents sweat evaporation. The Laplace pressure difference between the cross-scale fibers facilitates the management system to ultimately expulse sweat. This results in microfluidic control of sweat without the need for external forces, resulting in rapid (<5 s), sensitive (19.8 nA µM-1), and stable (with signal noise and drift suppression) sweat detection. This yarn sensor can be easily integrated into various fabrics, enabling the creation of health monitoring smart garments. The garments maintain good monitoring performance even after 20 washes. This work provides a solution for designing smart yarns for high-precision, stable, and non-invasive health monitoring.

Multiplex reverse transcription recombinase polymerase amplification combined with lateral flow biosensor for simultaneous detection of three viral pathogens in cattle.

Bovine viral diarrhea virus (BVDV), bovine epidemic fever virus (BEFV), and bovine respiratory syncytial virus (BRSV) cause respiratory symptoms in cattle. The absence of rapid, precise, and easily accessible diagnostic methods poses difficulties for herders and veterinary epidemiologists during outbreaks of major infectious animal diseases. Considering the mixed infection of viruses, a multiple-detection method, reverse transcription recombinase polymerase amplification (mRT-RPA) combined with a lateral flow biosensor (LFB), was established to simultaneously detect the three pathogens. This technique is based on the specific binding of three differently labeled RT-RPA products (DNA sequences) to antibodies on the three test lines of the LFB, achieving multiplex detection through the presence or absence of coloration on the LFB test lines. The fluorescence values of the LFB test lines are recorded by a test strip reader. The mRT-RPA-LFB assay completes detection at a constant temperature of 41 °C within 33 min. The limits of detection (LODs) for BVDV, BEFV and BRSV were 2.62 × 101, 2.42 × 101 and 2.56 × 101 copies/µL, respectively. No cross-reactivity was observed with the other six bovine viruses. The developed method showed satisfactory intra- and inter-assay precision, and the average coefficients of variation were ranged from 2.92 % to 3.99 %. The diagnostic sensitivity and specificity were 98.11 % and 100 %, respectively, which were highly consistent with the RT-qPCR assay, and the kappa value was 0.988 (95 % confidence interval, CI). In general, the mRT-RPA-LFB assay has the potential to become a powerful tool for rapid screening of cattle diseases because of its advantages such as fast detection speed, convenient operation, strong specificity, and high sensitivity.

Protein-driven interaction enhanced electrochemiluminescence biosensor of hydrogen-bonded biohybrid organic frameworks for sensitive immunoassay of disease markers.

The oriented design of reticular materials as emitters can significantly enhance the sensitivity of electrochemiluminescence (ECL) sensing analysis for disease markers. However, due to the structural fragility of hydrogen bonds, relational research on hydrogen-bonded organic frameworks (HOFs) has not been thoroughly conducted. Additionally, the modulation of luminescence behavior through HOFs has been rarely reported. In view of this, hydrogen-bonded biohybrid organic frameworks (HBOFs) were synthesized and recruited for ECL immunoassay applications. HBOFs was easily prepared using 6,6',6″,6‴-(pyrene-1,3,6,8-tetrayl)tetrakis(2-naphthoic acid) as linkers via bovine serum albumin (BSA) activated hydrogen-bonded cross-linking. The material exhibited good fluorescence emission characteristics. And the highly ordered topological structure and molecular motion limitation mediated by BSA overcome aggregation-caused quenching and generate strong aggregation induced emission, expressing hydrogen-bond interaction enhanced ECL (HIE-ECL) activity with the participation of tri-n-propylamine. Furthermore, a sandwich immunosensor was constructed employing cobalt-based metal-phenolic network (CMPN) coated ferrocene nanoparticles (FNPs) as quenchers (CMPN@FNPs). Signal closure can be achieved by annihilating the excited state through electron transfer from both CMPN and FNPs. Using a universal disease marker, carcinoembryonic antigen, as the analysis model, the signal-off sensor obtained a detection limit of 0.47 pg/mL within the detection range of 1 pg/mL - 50 ng/mL. The synthesis and application of highly stable HBOFs triggered by proteins provide a reference for the development of new reticular ECL signal labels, and electron transfer model provides flexible solutions for more sensitive sensing analysis.

Graphitic carbon nitride-based electrochemical sensors: A comprehensive review of their synthesis, characterization, and applications.

Graphitic carbon nitride (g-C3N4) has garnered much attention as a promising 2D material in the realm of electrochemical sensors. It contains a polymeric matrix that can serve as an economical and non-toxic electrode material for the detection of a diverse range of analytes. However, its performance is impeded by a relatively limited active surface area and inherent instability. Although electrochemistry involving metal-doped g-C3N4 nanomaterials is rapidly progressing, it remains relatively unexplored. The metal doping of g-C3N4 augments the electrochemically active surface area of the resulting electrode, which has the potential to significantly enhance electrode kinetics and bolster catalytic activity. Consequentially, the main objective of this review is to provide insight into the intricacies of synthesizing and characterizing metal-doped g-C3N4. Furthermore, we comprehensively delve into the fundamental attributes of electrochemical sensors based on metal-doped g-C3N4, with a specific focus on healthcare and environmental applications. These applications encompass a meticulous exploration of detecting biomolecules, drug molecules, and organic pollutants.

Metal Halide Perovskite Nanocrystals-Intermediated Hydrogel for Boosting the Biosensing Performance.

Metal-halide perovskites have become attractive nanomaterials for advanced biosensors, yet the structural design remains challenging due to the trade-off between environmental stability and sensing sensitivity. Herein, a trinity strategy is proposed to address this issue by integrating Mn (II) substitution with CsPb2Cl5 inert shell and NH2-PEG-COOH coating for designing Mn2+-doped CsPbCl3/CsPb2Cl5 core/shell hetero perovskite nanocrystals (PMCP PNCs). The trinity strategy isolates the emissive Mn2+-doped CsPbCl3 core from water and the Mn2+ d-d transition generates photoluminescence with a long lifetime, endowing the NH2-PEG-COOH capped Mn2+-doped CsPbCl3/CsPb2Cl5 PNCs with robust water stability and oxygen-sensitive property. Given the structural integration, photoluminescent hydrogel biosensors are designed by embedding the PMCP PNCs into the hydrogel system to deliver on-site pesticide information on food products. Impressively, benefiting from the dual enzyme triggered-responsive property of PMCP PNCs, the hydrogel biosensor is endowed with ultra-high sensitivity toward chlorpyrifos pesticide at the nanogram per milliliter level. Such a robust PMCP PNCs-based hydrogel sensor can provide accurate pesticide information while guiding the construction of photoluminescent biosensors for upcoming on-site applications.

Sensitive colorimetric immunosensor using AuNP-functionalized polymer film for picogram-level detection of Tau protein intermediate aggregates.

Here we demonstrate for the first time that an antibody-gold nanoparticles (AuNPs)-polymer conjugate thin-film biosensor can easily be fabricated to selectively capture Tau protein. Gold nanoparticles (AuNPs) are employed as sensing elements, thus capitalizing on their propensity to undergo assembly or disassembly in response to the adsorption or conjugation of various biomolecules on their surface, thereby forming robust interactions with the target analyte. We show that the Tau protein in its different aggregation phases can be detected, by restricting the reaction area on the solid thin polymer film and thus reducing the diffusion effects usually encountered in immunosensors. A limit of detection (LOD) of 460 pg/mL was reached, demonstrating a great potential for detecting Tau in aggregation states. This sensor based on thin polymer film could open new routes for sensing and monitoring Tau protein in biological assays and biomedical diagnosis.

Electrochemical detection of MMP-2 with enhanced sensitivity: Utilizing T7 RNA polymerase and CRISPR-Cas12a for neuronal studies.

This study introduces a novel electrochemical biosensor for detecting Matrix Metalloproteinase-2 (MMP-2), a key biomarker in cancer diagnostics and tissue remodeling. The biosensor is based on a dual-amplification strategy utilizing T7 RNA polymerase isothermal amplification and CRISPR-Cas12a technology. The principle involves the release of a DNA template in the presence of MMP-2, leading to RNA synthesis by T7 RNA polymerase. This RNA activates CRISPR-Cas12a, which cleaves a DNA probe on the electrode surface, resulting in a measurable electrochemical signal.The biosensor demonstrated exceptional sensitivity, with a detection limit of 2.62 fM for MMP-2. This high sensitivity was achieved through the combination of transcriptional amplification and the collateral cleavage activity of CRISPR-Cas12a, which amplifies the signal. The sensor was able to detect MMP-2 across a wide dynamic range from 2 fM to 1 nM, showing a strong linear correlation between MMP-2 concentration and the electrochemical signal. In practical applications, the biosensor accurately detected elevated levels of MMP-2 in cell culture supernatants from HepG2 liver cancer cells, distinguishing them from normal LO2 liver cells. The use of an MMP-2 inhibitor confirmed the specificity of the detection. These results underscore the biosensor's potential for clinical diagnostics, particularly in early cancer detection and monitoring of tissue remodeling activities. The biosensor's design allows for rapid, point-of-care testing without the need for complex laboratory equipment, making it a promising tool for personalized healthcare and diagnostic applications.

Comparative analysis of electrophoresis and interferometric optical detection method for molecular weight determination of proteins.

Analytical detection methods play a pivotal role in scientific research, enabling the identification and quantification of specific analytes in various disciplines. This scientific report aims to compare two very different methodologies for determining the Molecular Mass (MM, also known as Molecular Weight, MW) of proteins: electrophoresis gel and the Interferometric Optical Detection Method (IODM). For this purpose, several proteins with different MM were selected. The electrophoresis technique was employed to validate the structure and MM of different parts or fragments of the Matrix Metallopeptidase 9 antibody (anti-MMP9), antibody against S100 calcium binding protein A6 (anti-S100A6) and Cystatin S4 antibody (anti-CST4) by examining the presence of bands with expected sizes. The IODM was applied to study the above-mentioned proteins (part of the antibodies) together with the protein G, as a reference to correlate the MM and protein sizes with the measured signal. We report the evidence of IODM as a competitive analytical approach for the determination of the MM of proteins for the first time. This innovative method allows for accurate MM determination using minimal sample volumes and concentrations, employing a simple experimental procedure that eliminates the requirement for protein denaturation.

Designing an Electrochemical Biosensor Based on Voltammetry for Measurement of Human Chorionic Gonadotropin.

Background: Human chorionic gonadotropin (hCG) is a polypeptide hormone synthesized during pregnancy and is also upregulated in some pathologic conditions such as certain tumors. Its measurement is essential for diagnosing pregnancy and malignancies. Despite numerous attempts to introduce an accurate method capable of detecting hCG levels, several limitations are found in previous techniques. This study aimed to address the limitations of current hCG assay methods by designing an electrochemical biosensor based on voltammetry for the rapid, selective, inexpensive, and sensitive measurement of hCG levels. Methods: A carbon paste electrode was prepared and functionalized by para-aminobenzoic acid. The primary anti-ß-hCG monoclonal antibody was immobilized on the electrode surface by activating the carboxyl groups with 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide and N-hydroxysuccinimide solutions. The study also involved optimizing parameters such as the time for primary antibody fixation, the time for hCG attachment, and the pH of the hydrogen peroxide solution to maximize the biosensor response. Different concentrations of hCG hormone were prepared and loaded on the electrode surface, the secondary antibody labeled with HRP enzyme was applied, thionine in phosphate-buffered saline solution was placed on the electrode surface, and the differential pulse electrical signal was recorded. Results: The linear range ranged from 5 to 100 mIU/ml, and the limit of detection was calculated as 0.11 mIU. The relative standard deviation was 3% and 2% for five repeated measurements of commercial standard samples with concentrations of 2 and 20 mIU/mL, respectively. The percent recovery was obtained from 98.3% to 101.5%. Conclusion: The sensor represents a promising advancement in hCG level measurement, offering a potential solution to overcome the existing limitations in current diagnostic strategies. Simple and inexpensive design, detecting hCG in its important clinical range during early pregnancy, and successful measurement of hCG in real serum samples are the advantages of this sensor.

Cerium oxide mimetic enzyme based colorimetric detection of potential oesophageal cancer biomarkers.

Oesophageal cancer (OC) is a prevalent malignant tumor that poses a significant threat to individuals. Current mainstream detection method is endoscopy, which requires professional operators and expensive instruments. Therefore, it is crucial to develop a rapid, easy-to-operate, and low-cost detection method. In this study, an RNA colorimetric biosensor was successfully constructed using cerium oxide mimetic enzyme. The sensor is constructed on 96-well plates, which are immobilized with DNA-RNA-DNA complexes in microtiter wells when target RNA is present. This immobilization is based on the principle of base complementary pairing. The CeO2 immobilized has the unique advantage of catalyzing the bluing of 3,3',5,5'-tetramethylbenzidine (TMB) directly without the need any additional oxidant in microtiter wells. This property allows for the detection of RNA and enables the visualization of multiple sample assays. Furthermore, the RNA colorimetric sensor demonstrates good selectivity, immunity to interference, and high stability. Under optimal conditions, the sensor exhibited linearity in the range of 10-13 to 10-9 M with a detection limit of 33.26 fM. Therefore, this study presents a new detection method for oesophageal cancer screening.

Diagnosis of viral infectious diseases through sensitive detection of human serum antibodies using a modified label-free electrochemical biosensor.

Rapid virus identification is crucial for preventing outbreaks. The COVID-19 pandemic has highlighted the critical nature of rapid virus detection. Here, we designed a label-free electrochemical biosensor modified with gold nanoparticles (AuNPs) to detect IgG antibodies from human serum, enabling rapid point-of-care diagnostics. AuNPs were synthesized and characterized. A multivariate optimization was carried out to determine the optimal condition for functionalizing AuNPs with anti-IgG. Subsequently, using a glassy carbon electrode (GCE), a modified AuNPs/GCE electrochemical biosensor was developed for IgG detection. The results indicated that AuNPs displayed a spherical morphology with a size distribution of 19.54 nm. Additionally, the zeta potential was recorded at -7.84 mV. Central composite design (CCD) analysis determined the optimal conditions for functionalizing AuNPs to be an anti-IgG concentration of 320 µg mL-1, a temperature of 25 °C, and pH of 7.4. The characterization study confirmed the successful synthesis and functionalization of AuNPs. Through electrochemical impedance spectroscopy measurement, the biosensor demonstrated a limit of detection (LOD) of 0.2 ng mL-1 and limit of quantification (LOQ) of 0.8 ng mL-1. Furthermore, tests in real samples showed the interaction between IgG antibodies in serum samples and AuNPs/GCE, confirming the biosensor's ability to detect and quantify IgG in clinical samples.

A Modular Biosensor Design for Quantitative Measurement of Free Nedd8.

The objective of our study was to develop a genetically encoded biosensor for quantification of Nedd8, a post-translational modifier that regulates cellular signals through conjugation to other proteins. Perturbations in the balance of free (i.e., unconjugated) and conjugated Nedd8 caused by defects in Nedd8 enzymes or cellular stress are implicated in various diseases. Despite the biological and biomedical importance of Nedd8 dynamics, no method exists for direct quantification of free Nedd8, hindering the study of Nedd8 and activities of its associated enzymes. Genetically encoded biosensors are established as tools to study other dynamic systems, but limitations of current biosensor design methods make them poorly suited for free Nedd8 quantification. We have developed a modular method to design genetically encoded biosensors that employs a target binding domain and two reporter domains positioned on opposite sides of the target binding site. Target quantification is based on competition between target binding and the interaction of the reporter domains. We applied our design strategy to free Nedd8 quantification by developing a selective binder for free Nedd8 and combining it with fluorescent or split nanoluciferase reporters. Our sensors produced quantifiable and specific signals for free Nedd8 and enabled real-time monitoring of deneddylation by DEN1 with a physiological substrate. Our sensor design will be useful for high-throughput screening for deneddylation inhibitors, which have potential in treatment of cancers such as acute lymphoblastic leukemia. The modular design strategy can be extended to develop genetically encoded quantitative biosensors for other proteins of interest.