Upconverting Nanoparticle-based Enhanced Luminescence Lateral-Flow Assay for Urinary Biomarker Monitoring.

Development of efficient portable sensors for accurately detecting biomarkers is crucial for early disease diagnosis, yet remains a significant challenge. To address this need, we introduce the enhanced luminescence lateral-flow assay, which leverages highly luminescent upconverting nanoparticles (UCNPs) alongside a portable reader and a smartphone app. The sensor's efficiency and versatility were shown for kidney health monitoring as a proof of concept. We engineered Er3+- and Tm3+-doped UCNPs coated with multiple layers, including an undoped inert matrix shell, a mesoporous silica shell, and an outer layer of gold (UCNP@mSiO2@Au). These coatings synergistically enhance emission by over 40-fold and facilitate biomolecule conjugation, rendering UCNP@mSiO2@Au easy to use and suitable for a broad range of bioapplications. Employing these optimized nanoparticles in lateral-flow assays, we successfully detected two acute kidney injury-related biomarkers─kidney injury molecule-1 (KIM-1) and neutrophil gelatinase-associated lipocalin (NGAL)─in urine samples. Using our sensor platform, KIM-1 and NGAL can be accurately detected and quantified within the range of 0.1 to 20 ng/mL, boasting impressively low limits of detection at 0.28 and 0.23 ng/mL, respectively. Validating our approach, we analyzed clinical urine samples, achieving biomarker concentrations that closely correlated with results obtained via ELISA. Importantly, our system enables biomarker quantification in less than 15 min, underscoring the performance of our novel UCNP-based approach and its potential as reliable, rapid, and user-friendly diagnostics.

1D topological photonic crystal based nanosensor for tuberculosis detection.

In this study, we present a nanosized biosensor based on the photobiological properties of one-dimensional (1D) topological photonic crystals (PCs). A topological structure had been designed by combining two PC structures (PC 1 and PC 2) comprised of functional material layers, Si and SiO2. These two, PC 1 and PC 2, differ in terms of the thickness and arrangement of these dielectric materials. We carried out a comparison between two distinct topological PCs: one using random PCs, and the other featuring a mirror heterostructure. Tuberculosis may be diagnosed by inserting a sensor layer into 1D topological PCs. The sensing process is based on the refractive indexes of the analytes in the sensor layer. When the 1D-topological heterostructure-based PC and its mirror-image structures are stacked together, the sensor becomes more efficient for analyte detection than the conventional PCs. The random-based topological PC outperformed the heterostructure-based topological PC in analyte sensing. Photonic media witness notable blue shifts due to the analytes' variations in refractive index. The numerical results of the sensor are computed using the transfer matrix approach. Effective results are achieved by optimizing the thicknesses of the sensor layer and dielectric layers; number of periods and incident angle. In normal incident light, the developed sensor shows a high sensitivity of 1500 nm RIU-1with a very low limit of detection in the order of 2.2 × 10-06RIU and a high-quality factor of 30 659.54.

The Horizons of Medical Mineralogy: Structure-Bioactivity Relationship and Biomedical Applications of Halloysite Nanoclay.

Medical mineralogy explores the interactions between natural minerals and living organisms such as cells, tissues, and organs and develops therapeutic and diagnostic applications in drug delivery, medical devices, and healthcare materials. Many minerals (especially clays) have been recognized for pharmacological activities and therapeutic potential. Halloysite clay (Chinese medicine name: Chishizhi), manifested as one-dimensional aluminum silicate nanotubes (halloysite nanotubes, HNTs), has gained applications in hemostasis, wound repair, gastrointestinal diseases, tissue engineering, detection and sensing, cosmetics, and daily chemicals formulations. Various biomedical applications of HNTs are derived from hollow tubular structures, high mechanical strength, good biocompatibility, bioactivity, and unique surface characteristics. This natural nanomaterial is safe, abundantly available, and may be processed with environmentally safe green chemistry methods. This review describes the structure and physicochemical properties of HNTs relative to bioactivity. We discuss surface area, porosity and surface defects, hydrophilicity, heterogeneity and charge of external and internal surfaces, as well as biosafety. The paper provides comprehensive guidance for the development of this tubule nanoclay and its advanced biomedical applications for clinical diagnosis and therapy.

A novel phosphotriesterase hybrid nanoflower-hydrogel sensor equipped with a smartphone detector for real-time on-site monitoring of organophosphorus pesticides.

Designing efficient and rapid methods for the detection of organophosphorus pesticides (OPs) residue is a prerequisite to mitigate their negative health impacts. In this study, we propose the concept of an enzyme catalysis system-based hydrogel kit integrated with a smartphone detector for in-field screening of OPs. Here, we rapidly prepared phosphotriesterase hybrid nanoflowers (PTE-HNFs) using a self-assembly strategy by adding external energy and embedded the nanocomposite in sodium alginate (SA) hydrogel to construct a target-responsive hydrogel kit. The color response of the kit is induced by catalyzing methyl parathion (MP) to produce p-nitrophenol. For on-site quantification, the color variations of the portable kit are converted into digital information through a smartphone, which exhibits an applicable linear range towards OPs. The hydrogel sensing platform demonstrates a wide linear range (1-150 µM) and low detection limit (0.15 µM) for MP while maintaining high reliability, excellent long-term stability, and ease of operation. Overall, the PTE-HNFs-based SA hydrogel kit provides a useful strategy for simple and sensitive detection of MP and holds great potential for applications in detecting OPs in food and environmental water.

Modular Fabrication of Microfluidic Graphene FET for Nucleic Acids Biosensing.

Graphene field-effect transistors (GFETs) are widely used in biosensing due to their excellent properties in biomolecular signal amplification, exhibiting great potential for high-sensitivity and point-of-care testing in clinical diagnosis. However, difficulties in complicated fabrication steps are the main limitations for the further studies and applications of GFETs. In this study, a modular fabrication technique is introduced to construct microfluidic GFET biosensors within 3 independent steps. The low-melting metal electrodes and intricate flow channels are incorporated to maintain the structural integrity of graphene and facilitate subsequent sensing operations. The as-fabricated GFET biosensor demonstrates excellent long-term stability, and performs effectively in various ion environments. It also exhibits high sensitivity and selectivity for detecting single-stranded nucleic acids at a 10 fm concentration. Furthermore, when combined with the CRISPR/Cas12a system, it facilitates amplification-free and rapid detection of nucleic acids at a concentration of 1 fm. Thus, it is believed that this modular-fabricated microfluidic GFET may shed light on further development of FET-based biosensors in various applications.

Nano-Photonic Crystal D-Shaped Fiber Devices for Label-Free Biosensing at the Attomolar Limit of Detection.

Maintaining both high sensitivity and large figure of merit (FoM) is crucial in regard to the performance of optical devices, particularly when they are intended for use as biosensors with extremely low limit of detection (LoD). Here, a stack of nano-assembled layers in the form of 1D photonic crystal, deposited on D-shaped single-mode fibers, is created to meet these criteria, resulting in the generation of Bloch surface wave resonances. The increase in the contrast between high and low refractive index (RI) nano-layers, along with the reduction of losses, enables not only to achieve high sensitivity, but also a narrowed resonance bandwidth, leading to a significant enhancement in the FoM. Preliminary testing for bulk RI sensitivity is carried out, and the effect of an additional nano-layer that mimics a biological layer where binding interactions occur is also considered. Finally, the biosensing capability is assessed by detecting immunoglobulin G in serum at very low concentrations, and a record LoD of 70 aM is achieved. An optical fiber biosensor that is capable of attaining extraordinarily low LoD in the attomolar range is not only a remarkable technical outcome, but can also be envisaged as a powerful tool for early diagnosis of diseases.

[Research progress in biosensors for adiponectin].

Adiponectin, a cytokine associated with adipose tissue, is a recently defined adipocytokine involved in insulin, glucose, and adipocyte metabolism. Reduced adiponectin levels can increase the risk of developing metabolic syndrome (MS). Adiponectin is considered an important target for the treatment of type 2 diabetes mellitus (T2DM) and MS due to its anti-atherosclerotic and insulin-sensitizing effects. Therefore, the accurate determination of adiponectin concentrations in human plasma is necessary for the management of both T2DM and MS. A variety of biosensors have been developed for the detection of biomarkers such as adiponectin. This paper reviews the applications of electrochemical sensors, surface-enhanced Raman scattering sensors, and microfluidic chip-based chemiluminescence sensors in the detection of adiponectin and the recent research progress in the sensors for the detection of adiponectin, aiming to provide a reference for the research and application of sensors for adiponectin in the medical field.

A novel dual-recognition fluorescent biosensor for sialyl-Lewisx sensitive detection.

Sialyl-Lewisx (SLex) is a tetrasugar, which plays an important role in initial inflammation and cancer cell metastasis, and can be used as a marker for cancer diagnosis and prognosis or a therapeutic target. Detecting SLex from complex biological media remains a significant challenge. Herein, a single-stranded DNA aptamer of SLex was screened based on the double-stranded DNA library-modified magnetic bead (MB)-SELEX technology. After 14 rounds of screening, 12,639 sequences were obtained and divided into nine families. Three representative sequences were selected based on the number of sequence repeats and Gibbs binding free energy, and the aptamer SLex-Apt2 with 80 nt length (Kd = 23.01 nM) had the best affinity and relatively high specificity for targeting SLex. Then, a novel dual-recognition fluorescent biosensor for SLex-sensitive detection based on aptamer SLex-Apt2 bio-dots and 3-aminobenzoboric acid-modified MB was developed. This method can detect SLex as low as 32 µM and has a good linear response in the range 100 µM to 2 mM. It has the advantages of low preparation cost, good targeting, and avoiding the occurrence of false-positive and false-negative detection results, which makes the biosensor more valuable in biological detection and clinical diagnosis.

Quantitative detection of pre-ovulatory luteinizing hormone surges in urine using the microfluidic vertical agitation approach.

Identifying the time of ovulation is an important process for women seeking and avoiding pregnancy. Luteinizing hormone (LH) plays an important role in ovulation, which is very important in the reproductive mechanism. Therefore, detecting the LH level is of great importance in monitoring ovulation. In this study, sensitive, rapid and selective electrochemical biosensors were developed to detect LH quantitatively from human urine samples and to monitor the ovulation period. Isopotential region and current density optimization studies revealed that sensors with an electrode width and spacing of 1 mm had the optimum performance. Electrochemical impedance spectra evidenced immobilization of DSP self-assembled monolayers and anti-LH-beta antibody on the surface. While the mobile phone vibrator led to a 3.5-fold enhancement in response signals, the agitation system developed resulted in a 10-fold improvement. The sensors displayed detection limits of 1.02 and 1.53 mIU/ml in the range of 0-40 mIU/ml LH concentration obtained using two statistical approaches. Additionally, the sensors showed no cross-reactivity to hCG, which is very similar in structure and is widely reported to have high cross-reactivity.

Ultrafast and highly sensitive detection of SARS-CoV-2 spike protein by field-effect transistor graphene-based biosensors.

The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), etiological agent for the coronavirus disease 2019 (COVID-19), has resulted in over 775 million global infections. Early diagnosis remains pivotal for effective epidemiological surveillance despite the availability of vaccines. Antigen-based assays are advantageous for early COVID-19 detection due to their simplicity, cost-effectiveness, and suitability for point-of-care testing (PoCT). This study introduces a graphene field-effect transistor-based biosensor designed for high sensitivity and rapid response to the SARS-CoV-2 spike protein. By functionalizing graphene with monoclonal antibodies and applying short-duration gate voltage pulses, we achieve selective detection of the viral spike protein in human serum within 100 µs and at concentrations as low as 1 fg/mL, equivalent to 8 antigen molecules per µL of blood. Furthermore, the biosensor estimates spike protein concentrations in serum from COVID-19 patients. Our platform demonstrates potential for next-generation PoCT antigen assays, promising fast and sensitive diagnostics for COVID-19 and other infectious diseases.

An overview of signal amplification strategies and construction methods on phage-based biosensors.

Phages are a class of viruses that specifically infect host bacteria. Compared to other recognition elements, phages offer several advantages such as high specificity, easy to obtain and good environmental tolerance, etc. These advantages underscore the potential of phages as recognition elements in the construction of biosensors. Therefore, the phage-based biosensors are currently garnering widespread attention for detecting pathogens in recent years. However, the test performance such as detection limit, sensitivity and stability of exicting phage-based biosensors require enhancement. In the design of sensors, the selection of various materials and construction methods significantly influences the test performance of the sensor, and employing appropriate signal amplification strategies and construction methods to devise biosensors based on different principles is an effective strategy to enhance sensor performance. The manuscript primarily focuses on the signal amplification strategies and construction methods employed in phage-based biosensors recent ten years, and summarizes the advantages and disadvantages of different signal amplification strategies and construction methods. Meanwhile, the manuscript discusses the relationship between sensor performance and various materials and construction methods, and reviews the application progress of phage-based electrochemical biosensors in the detection of foodborne bacteria. Furthermore, the manuscript points out the present limitations and the future research direction for the field of phage-based biosensors, so as to provide the reference for developing high-performance phage-based biosensors.

Investigating and Engineering an 1,2-Propanediol-Responsive Transcription Factor-Based Biosensor.

Transcription factor (TF)-based biosensors have arisen as powerful tools in the advancement of metabolic engineering. However, with the emergence of numerous bioproduction targets, the variety of applicable TF-based biosensors remains severely limited. In this study, we investigated and engineered an 1,2-propanediol (1,2-PD)-responsive transcription activator, PocR, from Salmonella typhimurium to enrich the current biosensor repertoire. Heterologous characterization of PocR in E. coli revealed a significantly limited operational range and dynamic range, primarily attributed to the leaky binding between PocR and its corresponding promoters in the absence of the 1,2-PD inducer. Promiscuity characterization uncovered the minor responsiveness of PocR toward glycerol and 1,2-butanediol (1,2-BD). Using AlphaFold-predicted structure and protein mutagenesis, we preliminarily explored the underlying mechanism of PocR. Based on the investigated mechanism, we engineered a PcoR-F46R/G105D variant with an altered inducer specificity to glycerol, as well as a PocR-ARE (Q107A/S192R/A203E) variant with nearly a 4-fold higher dynamic range (6.7-fold activation) and a 20-fold wider operational range (0-20 mM 1,2-PD). Finally, we successfully converted PocR to a repressor through promoter engineering. Integrating the activation and repression functions established a versatile 1,2-PD-induced bifunctional regulation system based on PocR-ARE. Our work showcases the exploration and exploitation of an underexplored type of transcriptional activator capable of recruiting RNA polymerase. It also expands the biosensor toolbox by providing a 1,2-PD-responsive bifunctional regulator and glycerol-responsive activator.

A colorimetric/electrochemical microfluidic biosensor using target-triggered DNA hydrogels for organophosphorus detection.

Organophosphorus compounds are widely distributed and highly toxic to the environment and living organisms. The current detection of organophosphorus compounds is based on a single-mode method, which makes it challenging to achieve good portability, accuracy, and sensitivity simultaneously. This study designed a multifunctional microfluidic chip to develop a dual-mode biosensor employing a DNA hydrogel as a carrier and aptamers as recognition probes for the colorimetric/electrochemical detection of malathion, an organophosphorus compound. The biosensor balanced portability and stability by combining a microfluidic chip and target-triggered DNA hydrogel-sensing technologies. Moreover, the biosensor based on target-triggered DNA hydrogel modified microfluidic developed in this study exhibited a dual-mode response to malathion, providing both colorimetric and electrochemical signals. The colorimetric mode enables rapid visualization and qualitative detection and, when combined with a smartphone, allows on-site quantitative analysis with a detection limit of 56 nM. The electrochemical mode offers a broad linear range (0.01-3000 µM) and high sensitivity (a limit of detection of 5 nM). The two modes could validate each other and improve the accuracy of detection. The colorimetric/electrochemical dual-mode biosensor based on target-triggered DNA hydrogel modified microfluidic chip offers a portable, simple, accurate, and sensitive strategy for detecting harmful environmental and food substances.

Ligify: Automated Genome Mining for Ligand-Inducible Transcription Factors.

Prokaryotic transcription factors can be repurposed into biosensors for the ligand-inducible control of gene expression, but the landscape of chemical ligands for which biosensors exist is extremely limited. To expand this landscape, we developed Ligify, a web application that leverages information in enzyme reaction databases to predict transcription factors that may be responsive to user-defined chemicals. Candidate transcription factors are then incorporated into automatically generated plasmid sequences that are designed to express GFP in response to the target chemical. Our benchmarking analyses demonstrated that Ligify correctly predicted 31/100 previously validated biosensors and highlighted strategies for further improvement. We then used Ligify to build a panel of genetic circuits that could induce a 47-fold, 5-fold, 9-fold, and 27-fold change in fluorescence in response to D-ribose, L-sorbose, isoeugenol, and 4-vinylphenol, respectively. Ligify should enhance the ability of researchers to quickly develop biosensors for an expanded range of chemicals and is publicly available at https://ligify.groov.bio.

A sensitive aptasensor mediated by gold nanoparticles/metal organic framework lattice for detection of Pb2+ ion in marine products.

Herein, an enzyme-free fluorescent aptasensor was introduced for the ultrasensitive quantification of lead (Pb2+) ion as a hazardous pollutant of the environment and foodstuffs. A nanocomposite of zeolitic imidazolate frameworks-8 and gold nanoparticles (ZIF-8@AuNPs) was utilized as an efficient quencher of the fluorescence intensity of carboxyfluorescein (FAM) signal reporter. The establishment of a hybrid structure between attached aptamer on ZIF-8@AuNPs nanocomposite, and its FAM-tagged complementary (CP) strand decreased the fluorescence response. The preferential binding between the aptamer and Pb2+ released CP strands, which retrieved the fluorescence signal. The aptasensor could assess Pb2+ in the linear concentration range of 1 pM-1 nM with a detection limit (LOD) of 0.24 pM. Besides, it could quantify Pb2+ in various samples, including fish, shrimp, tap water, milk, and serum samples. The developed aptasensor with the superiorities of easiness, cost-effectiveness, easy-to-operate, and rapidness is promising for controlling marine foodstuff safety.

Cyanobacterial Phycocyanin-Based Electrochemical Biosensor for the Detection of the Free Radical Hydrogen Peroxide.

Cyanobacteria are photosynthetic prokaryotes that inhabit extreme environments by modifying their photosensitive chemoreceptors called cyanobacteriochromes (CBCRs) which are linear tetrapyrrole-linked phycobilin molecules. These light-sensitive phycobilin from Spirulina platensis is recognized as a potential photoreceptor tool in optogenetics for monitoring cellular morphogenesis. We prepared and extracted highly fluorescent cyanobacterial phycocyanin (C-PC) by irradiating the culture with ambient red light. The crude phycocyanin was subjected to ion exchange chromatography, and its purity was monitored using UV-visible, fluorescence, and FT-IR spectroscopy methods. In the conventional method, red light-induced C-PC exhibited strong antioxidant activity against H2O2, with 88.7% in vitro scavenging activity without requiring any other preservatives. Interestingly, this red light-acclimated phycocyanin was applied as a biosensing material for the detection of the free radical hydrogen peroxide (H2O2) using the enzyme horseradish peroxidase (HRP) as a mediator. The modified C-PC-HRP glassy carbon electrode (GCE) can detect H2O2 from 0.1 to 1600 µM. The lowest possible detection limit of the electrode for H2O2 was 19 nM. This electrode was used to detect free radical H2O2 in blood serum samples. The microstructure of the lyophilized PC under SEM showed a flat crystal pattern, which enabled the immobilization of HRP on the electrode surface and electron transfer.

Non-Faradaic Impedimetric Detection of Heavy Metal Ions via a Hybrid Nanoparticle-DNAzyme Biosensor.

Due to rapid industrialization, novel water-quality monitoring techniques for the detection of highly toxic and hazardous heavy metal ions are essential. Herein, a hybrid noble nanoparticle/DNAzyme electrochemical biosensor is proposed for the simultaneous and label-free detection of Pb2+ and Cr3+ in aqueous solutions. The sensor is based on the combination of a two-dimensional naked-platinum nanoparticle film and DNAzymes, whose double-helix configuration disassembles into smaller fragments in the presence of target-specific heavy metal ions. The electrochemical behavior of the fabricated sensor was investigated with non-faradaic electrochemical impedance spectroscopy (EIS), resulting in the successful detection of Pb2+ and Cr3+ well below their maximum permitted levels in tap water. So far, there has been no report on the successful detection of heavy metal ions utilizing the non-faradaic electrochemical impedance spectroscopy technique based on advanced nanomaterials paired with DNAzymes. This is also one of the few reports on the successful detection of chromium (III) via a sensor incorporating DNAzymes.

A 3D-Printed Do-It-Yourself ELISA Plate Reader as a Biosensor Tested on TNFα Assay.

Simple analytical devices suitable for the analysis of various biochemical and immunechemical markers are highly desirable and can provide laboratory diagnoses outside standard hospitals. This study focuses on constructing an easily reproducible do-it-yourself ELISA plate reader biosensor device, assembled from generally available and inexpensive parts. The colorimetric biosensor was based on standard 96-well microplates, 3D-printed parts, and a smartphone camera as a detector was utilized here as a tool to replace the ELISA method, and its function was illustrated in the assay of TNFα as a model immunochemical marker. The assay provided a limit of detection of 19 pg/mL when the B channel of the RGB color model was used for calibration. The assay was well correlated with the ELISA method, and no significant matrix effect was observed for standard biological samples or interference of proteins expected in a sample. The results of this study will inform the development of simple analytical devices easily reproducible by 3D printing and found on generally available electronics.

Label-Free Electrochemical Dopamine Biosensor Based on Electrospun Nanofibers of Polyaniline/Carbon Nanotube Composites.

The development of conducting polymer incorporated with carbon materials-based electrochemical biosensors has been intensively studied due to their excellent electrical, optical, thermal, physical and chemical properties. In this work, a label-free electrochemical dopamine (DA) biosensor based on polyaniline (PANI) and its aminated derivative, i.e., poly(3-aminobenzylamine) (PABA), composited with functionalized multi-walled carbon nanotubes (f-CNTs), was developed to utilize a conducting polymer as a transducing material. The electrospun nanofibers of the composites were fabricated on the surface of fluorine-doped tin oxide (FTO)-coated glass substrate under the optimized condition. The PANI/f-CNTs and PABA/f-CNTs electrospun nanofibers were characterized by attenuated total reflectance-Fourier transform infrared (ATR-FTIR) spectroscopy, X-ray photoelectron spectroscopy (XPS), scanning electron microscopy (SEM) and transmission electron microscopy (TEM), which confirmed the existence of f-CNTs in the composites. The electroactivity of the electrospun nanofibers was investigated in phosphate buffer saline solution using cyclic voltammetry (CV) before being employed for label-free electrochemical detection of DA using differential pulse voltammetry (DPV). The sensing performances including sensitivity, selectivity, stability, repeatability and reproducibility of the fabricated electrospun nanofiber films were also electrochemically evaluated. The electrochemical DA biosensor based on PANI/f-CNTs and PABA/f-CNTs electrospun nanofibers exhibited a sensitivity of 6.88 µA·cm-2·µM-1 and 7.27 µA·cm-2·µM-1 in the linear range of 50-500 nM (R2 = 0.98) with a limit of detection (LOD) of 0.0974 µM and 0.1554 µM, respectively. The obtained DA biosensor showed great stability, repeatability and reproducibility with precious selectivity under the common interferences, i.e., glucose, ascorbic acid and uric acid. Moreover, the developed electrochemical DA biosensor also showed the good reliability under detection of DA in artificial urine.

An Efficient Bio-Receptor Layer Combined with a Plasmonic Plastic Optical Fiber Probe for Cortisol Detection in Saliva.

Cortisol is a clinically validated stress biomarker that takes part in many physiological and psychological functions related to the body's response to stress factors. In particular, it has emerged as a pivotal tool for understanding stress levels and overall well-being. Usually, in clinics, cortisol levels are monitored in blood or urine, but significant changes are also registered in sweat and saliva. In this work, a surface plasmon resonance probe based on a D-shaped plastic optical fiber was functionalized with a glucocorticoid receptor exploited as a highly efficient bioreceptor specific to cortisol. The developed plastic optical fiber biosensor was tested for cortisol detection in buffer and artificial saliva. The biosensor response showed very good selectivity towards other hormones and a detection limit of about 59 fM and 96 fM in phosphate saline buffer and artificial saliva, respectively. The obtained detection limit, with a rapid detection time (about 5 min) and a low-cost sensor system, paved the way for determining the cortisol concentration in saliva samples without any extraction process or sample pretreatment via a point-of-care test.