Biosensors

19 pages, 3040 KiB

Open AccessArticle

Identification of Podoplanin Aptamers by SELEX for Protein Detection and Inhibition of Platelet Aggregation Stimulated by C-Type Lectin-like Receptor 2

by Hui-Ju Tsai, Kai-Wen Cheng, Jou-Chen Li, Tsai-Xiang Ruan, Ting-Hsin Chang, Jin-Ru Wang and Ching-Ping Tseng

Biosensors 2024, 14(10), 464; https://doi.org/10.3390/bios14100464 - 27 Sep 2024

Abstract

Tumor cell-induced platelet aggregation (TCIPA) is a mechanism for the protection of tumor cells in the bloodstream and the promotion of tumor progression and metastases. The platelet C-type lectin-like receptor 2 (CLEC-2) can bind podoplanin (PDPN) on a cancer cell surface to facilitate [...] Read more.

Tumor cell-induced platelet aggregation (TCIPA) is a mechanism for the protection of tumor cells in the bloodstream and the promotion of tumor progression and metastases. The platelet C-type lectin-like receptor 2 (CLEC-2) can bind podoplanin (PDPN) on a cancer cell surface to facilitate TCIPA. Selective blockage of PDPN-mediated platelet–tumor cell interaction is a plausible strategy for inhibiting metastases. In this study, we aimed to screen for aptamers, which are the single-stranded DNA oligonucleotides that form a specific three-dimensional structure, bind to specific molecular targets with high affinity and specificity, bind to PDPN, and interfere with PDPN/CLEC-2 interactions. The systematic evolution of ligands by exponential enrichment (SELEX) was employed to enrich aptamers that recognize PDPN. The initial characterization of ssDNA pools enriched by SELEX revealed a PDPN aptamer designated as A1 displaying parallel-type G-quadruplexes and long stem-and-loop structures and binding PDPN with a material with a dissociation constant (K_d) of 1.3 ± 1.2 nM. The A1 aptamer recognized both the native and denatured form of PDPN. Notably, the A1 aptamer was able to quantitatively detect PDPN proteins in Western blot analysis. The A1 aptamer could interfere with the interaction between PDPN and CLEC-2 and inhibit PDPN-induced platelet aggregation in a concentration-dependent manner. These findings indicated that the A1 aptamer is a candidate for the development of biosensors in detecting the levels of PDPN expression. The action by A1 aptamer could result in the prevention of tumor cell metastases, and if so, could become an effective pharmacological agent in treating cancer patients. Full article

(This article belongs to the Special Issue Design and Application of Novel Nucleic Acid Probe)

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13 pages, 2703 KiB

Open AccessArticle

Portable Electrochemical System and Platform with Point-of-Care Determination of Urine Albumin-to-Creatinine Ratio to Evaluate Chronic Kidney Disease and Cardiorenal Syndrome

by Shuenn-Yuh Lee, Ding-Siang Ciou, Hao-Yun Lee, Ju-Yi Chen, Yi-Chieh Wei and Meng-Dar Shieh

Biosensors 2024, 14(10), 463; https://doi.org/10.3390/bios14100463 - 27 Sep 2024

Abstract

Abstract: The urine albumin (Alb)-to-creatinine (Crn) ratio (UACR) is a sensitive and early indicator of chronic kidney disease (CKD) and cardiorenal syndrome. This study developed a portable and wireless electrochemical-sensing platform for the sensitive and accurate determination of UACR. The developed platform consists [...] Read more.

Abstract: The urine albumin (Alb)-to-creatinine (Crn) ratio (UACR) is a sensitive and early indicator of chronic kidney disease (CKD) and cardiorenal syndrome. This study developed a portable and wireless electrochemical-sensing platform for the sensitive and accurate determination of UACR. The developed platform consists of a carbon nanotube (CNT)-2,2′-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid)(ABTS)-based modified UACR sensor, a miniaturised potentiostat, a cup holder embedded with a magnetic stirrer and a smartphone app. The UACR sensing electrode is composed of two screen-printed carbon working electrodes, one screen-printed carbon counter electrode and a screen-printed AgCl reference electrode. The miniaturised potentiostat, which is controlled by the developed app, performs cyclic voltammetry and amperometry to detect Alb and Crn, respectively. Clinical trials of the proposed system by using spot urine samples from 30 diabetic patients indicate that it can accurately classify all three CKD risk statuses within 30 min. The high accuracy of our proposed sensing system exhibits satisfactory agreement with the commercial biochemical analyser TBA-25FR (Y = 0.999X, R² = 0.995). The proposed UACR sensing system offers a convenient, reliable and affordable solution for personal mobile health monitoring and point-of-care urinalysis. Full article

(This article belongs to the Special Issue Electrochemical Biosensors for Disease Detection)

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11 pages, 2694 KiB

Open AccessArticle

Microfluidic Detection Platform for Determination of Ractopamine in Food

by Cheng-Xue Yu, Kuan-Hsun Huang, To-Lin Chen, Chan-Chiung Liu and Lung-Ming Fu

Biosensors 2024, 14(10), 462; https://doi.org/10.3390/bios14100462 - 26 Sep 2024

Abstract

A novel microfluidic ractopamine (RAC) detection platform consisting of a microfluidic RAC chip and a smart analysis device is proposed for the determination of RAC concentration in meat samples. This technology utilizes gold nanoparticles (AuNPs) modified with glutamic acid (GLU) and polyethyleneimine (PEI) [...] Read more.

A novel microfluidic ractopamine (RAC) detection platform consisting of a microfluidic RAC chip and a smart analysis device is proposed for the determination of RAC concentration in meat samples. This technology utilizes gold nanoparticles (AuNPs) modified with glutamic acid (GLU) and polyethyleneimine (PEI) to measure RAC concentration in food products. When RAC is present, AuNPs aggregate through hydrogen bonding, causing noticeable changes in their optical properties, which are detected using a self-built UV–visible micro-spectrophotometer. Within the range of 5 to 80 ppb, a linear relationship exists between the absorbance ratio (A_693nm/A_518nm) (Y) and RAC concentration (X), expressed as Y = 0.0054X + 0.4690, with a high coefficient of determination (R² = 0.9943). This method exhibits a detection limit of 1.0 ppb and achieves results within 3 min. The practical utility of this microfluidic assay is exemplified through the evaluation of RAC concentrations in 50 commercially available meat samples. The variance between concentrations measured using this platform and those determined via liquid chromatography–tandem mass spectrometry (LC-MS/MS) is less than 8.33%. These results underscore the viability of the microfluidic detection platform as a rapid and cost-effective solution for ensuring food safety and regulatory compliance within the livestock industry. Full article

(This article belongs to the Special Issue Biosensors Based on Microfluidic Devices—2nd Edition)

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3 pages, 615 KiB

Open AccessCorrection

Correction: Hu et al. Ultrasensitive Silicon Nanowire Biosensor with Modulated Threshold Voltages and Ultra-Small Diameter for Early Kidney Failure Biomarker Cystatin C. Biosensors 2023, 13, 645

by Jiawei Hu, Yinglu Li, Xufang Zhang, Yanrong Wang, Jing Zhang, Jiang Yan, Junjie Li, Zhaohao Zhang, Huaxiang Yin, Qianhui Wei, Qifeng Jiang, Shuhua Wei and Qingzhu Zhang

Biosensors 2024, 14(10), 461; https://doi.org/10.3390/bios14100461 - 26 Sep 2024

Abstract

In the original publication [...] Full article

(This article belongs to the Special Issue Biosensors Based on Transistors)

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21 pages, 3297 KiB

Open AccessReview

Harnessing CRISPR/Cas Systems for DNA and RNA Detection: Principles, Techniques, and Challenges

by Heyjin Son

Biosensors 2024, 14(10), 460; https://doi.org/10.3390/bios14100460 - 26 Sep 2024

Abstract

The emergence of CRISPR/Cas systems has revolutionized the field of molecular diagnostics with their high specificity and sensitivity. This review provides a comprehensive overview of the principles and recent advancements in harnessing CRISPR/Cas systems for detecting DNA and RNA. Beginning with an exploration [...] Read more.

The emergence of CRISPR/Cas systems has revolutionized the field of molecular diagnostics with their high specificity and sensitivity. This review provides a comprehensive overview of the principles and recent advancements in harnessing CRISPR/Cas systems for detecting DNA and RNA. Beginning with an exploration of the molecular mechanisms of key Cas proteins underpinning CRISPR/Cas systems, the review navigates the detection of both pathogenic and non-pathogenic nucleic acids, emphasizing the pivotal role of CRISPR in identifying diverse genetic materials. The discussion extends to the integration of CRISPR/Cas systems with various signal-readout techniques, including fluorescence, electrochemical, and colorimetric, as well as imaging and biosensing methods, highlighting their advantages and limitations in practical applications. Furthermore, a critical analysis of challenges in the field, such as target amplification, multiplexing, and quantitative detection, underscores areas requiring further refinement. Finally, the review concludes with insights into the future directions of CRISPR-based nucleic acid detection, emphasizing the potential of these systems to continue driving innovation in diagnostics, with broad implications for research, clinical practice, and biotechnology. Full article

(This article belongs to the Special Issue CRISPR/Cas System-Based Biosensors)

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15 pages, 6187 KiB

Open AccessReview

Assessment of Microvascular Function Based on Flowmotion Monitored by the Flow-Mediated Skin Fluorescence Technique

by Andrzej Marcinek, Joanna Katarzynska, Katarzyna Cypryk, Agnieszka Los-Stegienta, Jolanta Slowikowska-Hilczer, Renata Walczak-Jedrzejowska, Jacek Zielinski and Jerzy Gebicki

Biosensors 2024, 14(10), 459; https://doi.org/10.3390/bios14100459 - 25 Sep 2024

Abstract

This review summarizes studies dedicated to the assessment of microvascular function based on microcirculatory oscillations monitored by the Flow-Mediated Skin Fluorescence (FMSF) technique. Two approaches are presented. The first approach uses oscillatory parameters measured under normoxic conditions, expressed as flowmotion (FM), vasomotion (VM), [...] Read more.

This review summarizes studies dedicated to the assessment of microvascular function based on microcirculatory oscillations monitored by the Flow-Mediated Skin Fluorescence (FMSF) technique. Two approaches are presented. The first approach uses oscillatory parameters measured under normoxic conditions, expressed as flowmotion (FM), vasomotion (VM), and the normoxia oscillatory index (NOI). These parameters have been used for the identification of impaired microcirculatory oscillations associated with intense physical exercise, post-COVID syndrome, psychological stress, and erectile dysfunction. The second approach involves characterization of the microcirculatory response to hypoxia based on the measurement of hypoxia sensitivity (HS). The HS parameter is used to characterize microvascular complications in diabetes, such as diabetic kidney disease and diabetic foot ulcers. Based on research conducted by the authors of this review, the FMSF parameter ranges characterizing microvascular function are presented. The diagnostic approach to assessing microvascular function based on flowmotion monitored by the FMSF technique has a wide range of applications and the potential to be integrated into widespread medical practice. Full article

(This article belongs to the Section Biosensors and Healthcare)

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15 pages, 3580 KiB

Open AccessArticle

NIR-Sensitive Squaraine Dye—Peptide Conjugate for Trypsin Fluorogenic Detection

by Priyanka Balyan, Shekhar Gupta, Sai Kiran Mavileti, Shyam S. Pandey and Tamaki Kato

Biosensors 2024, 14(10), 458; https://doi.org/10.3390/bios14100458 - 25 Sep 2024

Abstract

Trypsin enzyme has gained recognition as a potential biomarker in several tumors, such as colorectal, gastric, and pancreatic cancer, highlighting its importance in disease diagnosis. In response to the demand for rapid, cost-effective, and real-time detection methods, we present an innovative strategy utilizing [...] Read more.

Trypsin enzyme has gained recognition as a potential biomarker in several tumors, such as colorectal, gastric, and pancreatic cancer, highlighting its importance in disease diagnosis. In response to the demand for rapid, cost-effective, and real-time detection methods, we present an innovative strategy utilizing the design and synthesis of NIR-sensitive dye–peptide conjugate (SQ-3 PC) for the sensitive and selective monitoring of trypsin activity by fluorescence ON/OFF sensing. The current research deals with the design and synthesis of three unsymmetrical squaraine dyes SQ-1, SQ-2, and SQ-3 along with a dye–peptide conjugate SQ-3-PC as a trypsin-specific probe followed by their photophysical characterizations. The absorption spectral investigation conducted on both the dye alone and its corresponding dye–peptide conjugates in water, utilizing SQ-3 and SQ-3 PC respectively, reveals enhanced dye aggregation and pronounced fluorescence quenching compared to observations in DMSO solution. The absorption spectral investigation conducted on dye only and corresponding dye–peptide conjugates in water utilizing SQ-3 and SQ-3 PC, respectively, reveals not only the enhanced dye aggregation but also pronounced fluorescence quenching compared to that observed in the DMSO solution. The trypsin-specific probe SQ-3 PC demonstrated a fluorescence quenching efficiency of 61.8% in water attributed to the combined effect of aggregation-induced quenching (AIQ) and fluorescence resonance energy transfer (FRET). FRET was found to be dominant over AIQ. The trypsin-mediated hydrolysis of SQ-3 PC led to a rapid and efficient recovery of quenched fluorescence (5-fold increase in 30 min). Concentration-dependent changes in the fluorescence at the emission maximum of the dyes reveal that SQ-3 PC works as a trypsin enzyme-specific fluorescence biosensor with linearity up to 30 nM along with the limit of detection and limit of quantification of 1.07 nM and 3.25 nM, respectively. Full article

(This article belongs to the Special Issue Photonics for Bioapplications: Sensors and Technology)

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11 pages, 3081 KiB

Open AccessArticle

Ratiometric Electrochemical Detection of Interleukin-6 Using Electropolymerized Methylene Blue and a Multi-Walled Carbon-Nanotube-Modified Screen-Printed Carbon Electrode

by Zhuo Liu, Fengyu Liu, Chaofan Wang, Hongjuan Li, Yongqian Xu and Shiguo Sun

Biosensors 2024, 14(10), 457; https://doi.org/10.3390/bios14100457 - 25 Sep 2024

Abstract

Herein, we report a ratio-based electrochemical biosensor for the detection of interleukin-6 (IL-6). We electropolymerized methylene blue (MB) on the surface of screen-printed carbon electrodes; introduced an internal reference signal probe; modified the carboxylate multi-walled carbon nanotubes on the electrode surface to increase [...] Read more.

Herein, we report a ratio-based electrochemical biosensor for the detection of interleukin-6 (IL-6). We electropolymerized methylene blue (MB) on the surface of screen-printed carbon electrodes; introduced an internal reference signal probe; modified the carboxylate multi-walled carbon nanotubes on the electrode surface to increase the electrochemically active area; and finally linked the amino-modified IL-6 aptamer to the electrode surface through the Schiff base reaction, with bovine serum albumin (BSA) added to mask non-specific adsorption. After adding IL-6 to the samples, the signal of I_MB remained almost unchanged, while the signal of I[Fe(CN)₆]^3−/4− decreased with increasing IL-6 concentration. Thus, a novel ratiometric electrochemical sensor with a linear range of 0.001~1000.0 ng/mL and a low detection limit of 0.54 pg/mL was successfully developed. The sensor had high repeatability, stability, sensitivity, and practicability. It provides a new method for constructing proportional electrochemical sensors and detecting IL-6. Full article

(This article belongs to the Special Issue State-of-the-Art Biosensors in China (2nd Edition))

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13 pages, 3367 KiB

Open AccessArticle

Recovery and Analysis of Bacterial Membrane Vesicle Nanoparticles from Human Plasma Using Dielectrophoresis

by Jason P. Ware, Delaney K. Shea, Shelby L. Nicholas, Ella A. Stimson, Jessica L. Riesterer and Stuart D. Ibsen

Biosensors 2024, 14(10), 456; https://doi.org/10.3390/bios14100456 - 25 Sep 2024

Abstract

Bacterial membrane vesicle (BMV) nanoparticles are secreted naturally by bacteria throughout their lifecycle and are a rich source of biomarkers from the parent bacteria, but they are currently underutilized for clinical diagnostic applications, such as pathogen identification, due to the time-consuming and low-yield [...] Read more.

Bacterial membrane vesicle (BMV) nanoparticles are secreted naturally by bacteria throughout their lifecycle and are a rich source of biomarkers from the parent bacteria, but they are currently underutilized for clinical diagnostic applications, such as pathogen identification, due to the time-consuming and low-yield nature of traditional recovery methods required for analysis. The recovery of BMVs is particularly difficult from complex biological fluids. Here, we demonstrate a recovery method that uses dielectrophoretic (DEP) forces generated on electrokinetic microfluidic chips to isolate and analyze BMVs from human plasma. DEP takes advantage of the natural difference in dielectric properties between the BMVs and the surrounding plasma fluid to quickly and consistently collect these particles from as little as 25 µL of plasma. Using DEP and immunofluorescence staining of the LPS biomarker carried on BMVs, we have demonstrated a lower limit of detection of 4.31 × 10⁹ BMVs/mL. The successful isolation of BMVs from human plasma using DEP, and subsequent on-chip immunostaining for biomarkers, enables the development of future assays to identify the presence of specific bacterial species by analyzing BMVs from small amounts of complex body fluid. Full article

(This article belongs to the Special Issue Microfluidic Systems and Computational Imaging Methods in Lab-on-a-Chip Technologies)

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13 pages, 1758 KiB

Open AccessArticle

High Throughput Screening of Transcription Factor LysG for Constructing a Better Lysine Biosensor

by Qinggang Li, Haojie Ren, Zhenjiang Liao, Shuchang Xia and Xue Sun

Biosensors 2024, 14(10), 455; https://doi.org/10.3390/bios14100455 - 25 Sep 2024

Abstract

The biosensors based on transcription factors (TFs) are widely used in high throughput screening of metabolic overproducers. The unsatisfactory performances (narrow detection and dynamic ranges) of biosensors limit their practical application and need more improvement. In this study, using the TF LysG (sensing [...] Read more.

The biosensors based on transcription factors (TFs) are widely used in high throughput screening of metabolic overproducers. The unsatisfactory performances (narrow detection and dynamic ranges) of biosensors limit their practical application and need more improvement. In this study, using the TF LysG (sensing lysine) as an example, a biosensor optimization method was constructed by growth-coupled screening of TF random mutant libraries. The better the performance of the biosensor, the faster the strain grows under screening pressure. A LysG^{E15D, A54D, and I164V}-based biosensors were obtained, which were about 2-fold of the control in the detection and dynamic ranges. A lysine high-producer was screened effectively using the optimized biosensor with the production at 1.51 ± 0.30 g/L in flasks (2.22-fold of the original strain). This study provided a promising strategy for optimizing TF-based biosensors and was of high potential to be applied in the lysine high-producers screening process. Full article

(This article belongs to the Section Biosensor and Bioelectronic Devices)

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10 pages, 1706 KiB

Open AccessArticle

MNAzyme-Assisted Nucleic Acid Lateral Flow Assay for Cost-Effective, On-Site Mercury Detection

by Seok Hyeon Kim, Yujun Kim, Seokjoon Kim, Eun Sung Lee, Byung Seok Cha and Ki Soo Park

Biosensors 2024, 14(10), 454; https://doi.org/10.3390/bios14100454 - 25 Sep 2024

Abstract

Mercury ions (Hg²⁺) are toxic heavy metals present in the environment that pose significant health risks. An advanced detection system could allow for a prompt response and alleviate serious damage to humans. In this study, we developed a cost-effective, on-site detection [...] Read more.

Mercury ions (Hg²⁺) are toxic heavy metals present in the environment that pose significant health risks. An advanced detection system could allow for a prompt response and alleviate serious damage to humans. In this study, we developed a cost-effective, on-site detection method for Hg²⁺ using a multicomponent nucleic acid enzyme (MNAzyme)-assisted nucleic acid lateral flow assay (NALFA). The MNAzyme, which was engineered to contain thymine–thymine mismatches, is responsive only to the presence of Hg²⁺ and exerts efficient cleavage activity on substrates that can be captured by the NALFA strip, and thus the proposed system enables the visual detection of Hg²⁺ in the NALFA strip. Our assay demonstrated sufficient detection sensitivity and specificity to meet the WHO standards, offering a good practical alternative for rapid environmental and public health monitoring. Full article

(This article belongs to the Special Issue Probes for Biosensing and Bioimaging)

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42 pages, 14006 KiB

Open AccessReview

Black Silicon Surface-Enhanced Raman Spectroscopy Biosensors: Current Advances and Prospects

by Yaraslau Padrez and Lena Golubewa

Biosensors 2024, 14(10), 453; https://doi.org/10.3390/bios14100453 - 24 Sep 2024

Abstract

Black silicon was discovered by accident and considered an undesirable by-product of the silicon industry. A highly modified surface, consisting of pyramids, needles, holes, pillars, etc., provides high light absorption from the UV to the NIR range and gives black silicon its color—matte [...] Read more.

Black silicon was discovered by accident and considered an undesirable by-product of the silicon industry. A highly modified surface, consisting of pyramids, needles, holes, pillars, etc., provides high light absorption from the UV to the NIR range and gives black silicon its color—matte black. Although black silicon has already attracted some interest as a promising material for sensitive sensors, the potential of this material has not yet been fully exploited. Over the past three decades, black silicon has been actively introduced as a substrate for surface-enhanced Raman spectroscopy (SERS)—a molecule-specific vibrational spectroscopy technique—and successful proof-of-concept experiments have been conducted. This review focuses on the current progress in black silicon SERS biosensor fabrication, the recent advances in the design of the surface morphology and an analysis of the relation of surface micro-structuring and SERS efficiency and sensitivity. Much attention is paid to problems of non-invasiveness of the technique and biocompatibility of black silicon, its advantages over other SERS biosensors, cost-effectiveness and reproducibility, as well as the expansion of black silicon applications. The question of existing limitations and ways to overcome them is also addressed. Full article

(This article belongs to the Special Issue Advanced SERS Biosensors for Detection and Analysis)

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Biosensors, Volume 14, Issue 10 (October 2024) – 12 articles

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