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Bioassays and Biosensors for Rapid Detection and Analysis

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A special issue of Biosensors (ISSN 2079-6374). This special issue belongs to the section "Biosensors and Healthcare".

Deadline for manuscript submissions: closed (31 July 2023) | Viewed by 29251

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Prof. Dr. Zhen Zhang

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School of the Environment and Safety Engineering, Jiangsu University, Zhenjiang 212013, China
Interests: monoclonal antibody and nano-antibody of new pollutants; rapid analysis; immunoassays; biosensors and electrochemistry; signal amplification; cell imaging based on DNA functional materials
Special Issues, Collections and Topics in MDPI journals

Special Issue Information

Dear Colleagues,

The main topic of this Special Issue is the development of bioassays and biosensors and their applications for rapid detection and analysis, based on which the Special Issue is dedicated to collecting research articles that offer great innovation in principle design, technology and strategy application, and process simplification. Additionally, reviews reflecting current hotspots, new challenges, and future perspectives of bioanalysis and biosensor technology for rapid detection are particularly welcome.

Rapid detection and analysis require that the whole process from sample preparation to test results can be completed in a short time. It is usually simplified in sample treatment, test preparation, operational procedures and automation. Rapid detection and analysis technology, as a powerful tool for food safety monitoring, product quality control, toxic and harmful substance analysis, and acute disease diagnosis, is of great significance to improve the efficiency and strength of supervision, reduce the cost of sample testing, and ensure health safety and product quality.

With the rapid development of biotechnology in recent years, bioanalysis and biosensor technology has been widely applied in food, environment, medicine, military, and other fields. Due to its low cost, simple operation and equipment, quick analysis, and other advantages, it has great competitiveness in rapid detection and analysis. Moreover, various new technologies, such as nanotechnology, molecular imprinting, information science, and material science, provide a broad space for its development.

Prof. Dr. Zhen Zhang
Guest Editor

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Keywords

bioassays
biosensors
rapid detection
biomaterials
nanotechnology
immunoassays
wearable/portable devices
high throughput analysis

Published Papers (11 papers)

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Editorial

Jump to: Research, Review

2 pages, 178 KiB

Open AccessEditorial

Recent Progress in Biosensors Based on Biorecognition Molecules

by Zhen Zhang

Biosensors 2023, 13(9), 842; https://doi.org/10.3390/bios13090842 - 24 Aug 2023

Cited by 1 | Viewed by 644

Abstract

Biosensors are considered a popular technology to rapidly detect targets, and are generally composed of biorecognition molecules that specifically capture analytes and signal elements [...] Full article

(This article belongs to the Special Issue Bioassays and Biosensors for Rapid Detection and Analysis)

Research

Jump to: Editorial, Review

11 pages, 1584 KiB

Open AccessArticle

Biosensing Platform for the Detection of Biomarkers for ALI/ARDS in Bronchoalveolar Lavage Fluid of LPS Mice Model

by Nuha Khalid Alekhmimi, Dana Cialla-May, Qasem Ramadan, Shimaa Eissa, Jürgen Popp, Khaled Al-Kattan and Mohammed Zourob

Biosensors 2023, 13(7), 676; https://doi.org/10.3390/bios13070676 - 25 Jun 2023

Cited by 3 | Viewed by 1283

Abstract

Acute respiratory distress syndrome (ARDS) is a worldwide health concern. The pathophysiological features of ALI/ARDS include a pulmonary immunological response. The development of a rapid and low-cost biosensing platform for the detection of ARDS is urgently needed. In this study, we report the development of a paper-based multiplexed sensing platform to detect human NE, PR3 and MMP-2 proteases. Through monitoring the three proteases in infected mice after the intra-nasal administration of LPS, we showed that these proteases played an essential role in ALI/ARDS. The paper-based sensor utilized a colorimetric detection approach based on the cleavage of peptide–magnetic nanoparticle conjugates, which led to a change in the gold nanoparticle-modified paper sensor. The multiplexing of human NE, PR3 and MMP-2 proteases was tested and compared after 30 min, 2 h, 4 h and 24 h of LPS administration. The multiplexing platform of the three analytes led to relatively marked peptide cleavage occurring only after 30 min and 24 h. The results demonstrated that MMP-2, PR3 and human NE can provide a promising biosensing platform for ALI/ARDS in infected mice at different stages. MMP-2 was detected at all stages (30 min–24 h); however, the detection of human NE and PR3 can be useful for early- (30 min) and late-stage (24 h) detection of ALI/ARDS. Further studies are necessary to apply these potential diagnostic biosensing platforms to detect ARDS in patients. Full article

(This article belongs to the Special Issue Bioassays and Biosensors for Rapid Detection and Analysis)

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11 pages, 5438 KiB

Open AccessCommunication

Development and Validation of Aptasensor Based on MnO₂ for the Detection of Sulfadiazine Residues

by Xiaoling Zheng, Lulan Yang, Qi Sun, Lei Zhang and Tao Le

Biosensors 2023, 13(6), 613; https://doi.org/10.3390/bios13060613 - 03 Jun 2023

Cited by 3 | Viewed by 1261

Abstract

The monitoring of sulfadiazine (SDZ) is of great significance for food safety, environmental protection, and human health. In this study, a fluorescent aptasensor based on MnO₂ and FAM-labeled SDZ aptamer (FAM-SDZ30-1) was developed for the sensitive and selective detection of SDZ in food and environmental samples. MnO₂ nanosheets adsorbed rapidly to the aptamer through its electrostatic interaction with the base, providing the basis for an ultrasensitive SDZ detection. Molecular dynamics was used to explain the combination of SMZ1S and SMZ. This fluorescent aptasensor exhibited high sensitivity and selectivity with a limit of detection of 3.25 ng/mL and a linear range of 5–40 ng/mL. The recoveries ranged from 87.19% to 109.26% and the coefficients of variation ranged from 3.13% to 13.14%. In addition, the results of the aptasensor showed an excellent correlation with high-performance liquid chromatography (HPLC). Therefore, this aptasensor based on MnO₂ is a potentially useful methodology for highly sensitive and selective detection of SDZ in foods and environments. Full article

(This article belongs to the Special Issue Bioassays and Biosensors for Rapid Detection and Analysis)

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13 pages, 2738 KiB

Open AccessArticle

Enhanced Competitive Immunomagnetic Beads Assay Assisted with PAMAM-Gold Nanoparticles Multi-Enzyme Probes for Detection of Deoxynivalenol

by Kun Zeng, Jian Yang, Hao Su, Sheng Yang, Xinkai Gu, Zhen Zhang and Hongjun Zhao

Biosensors 2023, 13(5), 536; https://doi.org/10.3390/bios13050536 - 10 May 2023

Cited by 1 | Viewed by 1607

Abstract

Contamination of deoxynivalenol (DON) in grains has attracted widespread concern. It is urgently needed to develop a highly sensitive and robust assay for DON high-throughput screening. Antibody against DON was assembled on the surface of immunomagnetic beads orientationally by the aid of Protein G. AuNPs were obtained under the scaffolding of poly(amidoamine) dendrimer (PAMAM). DON-horseradish peroxidase (HRP) was combined on the periphery of AuNPs/PAMAM by a covalent link to develop DON-HRP/AuNPs/PAMAM. Magnetic immunoassay based on DON-HRP/AuNPs/PAMAM was optimized and that based on DON-HRP/AuNPs and DON-HRP was adopted as comparison. The limits of detection (LODs) were 0.447 ng/mL, 0.127 ng/mL and 0.035 ng/mL for magnetic immunoassays based on DON-HRP, DON-HRP/Au and DON-HRP/Au/PAMAM, respectively. Magnetic immunoassay based on DON-HRP/AuNPs/PAMAM displayed higher specificity towards DON and was utilized to analyze grain samples. The recovery for the spiked DON in grain samples was 90.8–116.2% and the method presented a good correlation with UPLC/MS. It was found that the concentration of DON was in the range of ND-3.76 ng/mL. This method allows the integration of dendrimer–inorganic NPs with signal amplification properties for applications in food safety analysis. Full article

(This article belongs to the Special Issue Bioassays and Biosensors for Rapid Detection and Analysis)

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7 pages, 492 KiB

Open AccessCommunication

A Comparison Study of the Detection Limit of Omicron SARS-CoV-2 Nucleocapsid by Various Rapid Antigen Tests

by Daniela Dobrynin, Iryna Polischuk and Boaz Pokroy

Biosensors 2022, 12(12), 1083; https://doi.org/10.3390/bios12121083 - 27 Nov 2022

Cited by 6 | Viewed by 2912

Abstract

Rapid antigen tests (RATs) are widely used worldwide to detect SARS-CoV-2 since they are an easy-to-use kit and offer rapid results. The RAT detects the presence of the nucleocapsid protein, which is located inside the virus. However, the sensitivity of the different RATs varies between commercially available kits. The test result might change due to various factors, such as the variant type, infection date, swab’s surface, the manner in which one performs the testing and the mucus components. Here, we compare the detection limit of seven commercially available RATs by introducing them to known SARS-CoV-2 nucleocapsid protein amounts from the Omicron variant. It allows us to determine the detection limit, disregarding the influences of other factors. A lower detection limit of the RAT is necessary since earlier detection will help reduce the spread of the virus and allow faster treatment, which might be crucial for the population at risk. Full article

(This article belongs to the Special Issue Bioassays and Biosensors for Rapid Detection and Analysis)

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12 pages, 1639 KiB

Open AccessArticle

Two-Way Detection of COVID-19 Spike Protein and Antibody Using All-Dielectric Metasurface Fluorescence Sensors

by Masanobu Iwanaga and Wanida Tangkawsakul

Biosensors 2022, 12(11), 981; https://doi.org/10.3390/bios12110981 - 07 Nov 2022

Cited by 6 | Viewed by 1698

Abstract

COVID-19 (or SARS-CoV-2) has deeply affected human beings worldwide for over two years, and its flexible mutations indicate the unlikeliness of its termination in a short time. Therefore, it is important to develop a quantitative platform for direct COVID-19 detection and human status monitoring. Such a platform should be simpler than nucleic acid amplification techniques such as polymerase chain reaction, and more reliable than the disposable test kits that are based on immunochromatography. To fulfill these requirements, we conducted proof-of-concept experiments for the quantitative detection of spike glycoprotein peptides and antibodies in one platform, i.e., all-dielectric metasurface fluorescence (FL) sensors. The high capability to enhance FL intensity enabled us to quantitatively measure the glycoproteins and antibodies more efficiently compared with the previous methods reported to date. Furthermore, the intrinsic limit of detection in the metasurface FL sensors was examined via confocal microscopy and found to be less than 0.64 pg/mL for glycoprotein peptides. Moreover, the sensors had a dynamic range more than five orders that of the target concentrations, indicating extremely high sensitivity. These two-way functions of the metasurface FL sensors can be helpful in reducing daily loads in clinics and in providing quantitative test values for proper diagnosis and cures. Full article

(This article belongs to the Special Issue Bioassays and Biosensors for Rapid Detection and Analysis)

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20 pages, 1994 KiB

Open AccessArticle

ImmunoDisk—A Fully Automated Bead-Based Immunoassay Cartridge with All Reagents Pre-Stored

by Benita Johannsen, Desirée Baumgartner, Lena Karkossa, Nils Paust, Michal Karpíšek, Nagihan Bostanci, Roland Zengerle and Konstantinos Mitsakakis

Biosensors 2022, 12(6), 413; https://doi.org/10.3390/bios12060413 - 14 Jun 2022

Cited by 4 | Viewed by 7541

Abstract

In this paper, we present the ImmunoDisk, a fully automated sample-to-answer centrifugal microfluidic cartridge, integrating a heterogeneous, wash-free, magnetic- and fluorescent bead-based immunoassay (bound-free phase detection immunoassay/BFPD-IA). The BFPD-IA allows the implementation of a simple fluidic structure, where the assay incubation, bead separation and detection are performed in the same chamber. The system was characterized using a C-reactive protein (CRP) competitive immunoassay. A parametric investigation on air drying of protein-coupled beads for pre-storage at room temperature is presented. The key parameters were buffer composition, drying temperature and duration. A protocol for drying two different types of protein-coupled beads with the same temperature and duration using different drying buffers is presented. The sample-to-answer workflow was demonstrated measuring CRP in 5 µL of human serum, without prior dilution, utilizing only one incubation step, in 20 min turnaround time, in the clinically relevant concentration range of 15–115 mg/L. A reproducibility assessment over three disk batches revealed an average signal coefficient of variation (CV) of 5.8 ± 1.3%. A CRP certified reference material was used for method verification with a concentration CV of 8.6%. Our results encourage future testing of the CRP-ImmunoDisk in clinical studies and its point-of-care implementation in many diagnostic applications. Full article

(This article belongs to the Special Issue Bioassays and Biosensors for Rapid Detection and Analysis)

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13 pages, 2788 KiB

Open AccessArticle

Rapid Capturing and Chemiluminescent Sensing of Programmed Death Ligand-1 Expressing Extracellular Vesicles

by Adeel Khan, Kaili Di, Haroon Khan, Nongyue He and Zhiyang Li

Biosensors 2022, 12(5), 281; https://doi.org/10.3390/bios12050281 - 28 Apr 2022

Cited by 7 | Viewed by 2430

Abstract

Cancer specific extracellular vesicles (EVs) are of significant clinical relevance, for instance programmed death ligand-1 (PD-L1) expressing EVs (PD-L1@EVs) have been shown to be ideal biomarker for non-invasive diagnosis of cancer and can predate the response of cancer patients to anti-PD-1/PD-L-1 immunotherapy. The development of sensitive and straightforward methods for detecting PD-L1@EVs can be a vital tool for non-invasive diagnosis of cancer. Most of the contemporary methods for EVs detection have limitations such as involvement of long and EV’s loss prone isolation methods prior to detection or they have employed expensive antibodies and instruments to accomplish detection. Therefore, we designed an ultracentrifugation-free and antibody-free sensing assay for PD-L1@EV by integrating Titanium oxide (TiO

_{2}

) coated magnetic beads (Fe

_{3}

_{4}

@TiO

_{2}

) rapid capturing of EVs from undiluted serum with aptamers specificity and chemiluminescence (CL) sensitivity. To accomplish this we used Fe

_{3}

_{4}

@TiO

_{2}

beads to rapidly capture EVs from the undiluted patient serum and added biotin labelled PD-L1 aptamer to specifically recognize PD-L1@EVs. Later, added streptavidin-modified Alkaline phosphates (ALP) taking advantage of biotin-streptavidin strong binding. Addition of CDP-star, a chemiluminescent substrate of ALP, initiates the chemiluminiscense that was recorded using spectrophotometer. The sensing assay showed high sensitivity with limit of detection (LOD) as low as

2.584 \times 10^{5}

EVs/mL and a wider linear correlation of CL intensity (a.u.) with the concentration of PD-L1@EVs from

10^{5}

10^{8}

EVs/mL. To examine the clinical utility of sensing assay we used undiluted serum samples from lung cancer patients and healthy individuals and successfully discern between healthy individuals and lung cancer patients. We are optimistic that the sensing assay can ameliorate our ability to be able to diagnose lung cancer non-invasively and can be helpful to predate the patient’s response to anti-PD-1/PD-L1 immunotherapy. Full article

(This article belongs to the Special Issue Bioassays and Biosensors for Rapid Detection and Analysis)

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9 pages, 2245 KiB

Open AccessCommunication

A Novel Enzyme-Free Ratiometric Fluorescence Immunoassay Based on Silver Nanoparticles for the Detection of Dibutyl Phthalate from Environmental Waters

by Hui Meng, Nannan Yao, Kun Zeng, Nuanfei Zhu, Yue Wang, Biying Zhao and Zhen Zhang

Biosensors 2022, 12(2), 125; https://doi.org/10.3390/bios12020125 - 16 Feb 2022

Cited by 7 | Viewed by 2577

Abstract

A novel ratiometric fluorescent immunoassay was developed based on silver nanoparticles (AgNPs) for the sensitive determination of dibutyl phthalate (DBP). In the detection system, AgNPs were labeled on the secondary antibody (AgNPs@Ab₂) for signal amplification, which aimed to regulate the H₂O₂ concentrations. When AgNPs-Ab₂ and antigen–primary antibody (Ab₁) were linked by specific recognition, the blue fluorescence of Scopoletin (SC) could be effectively quenched by the H₂O₂ added while the red fluorescence of Amplex Red (AR) was generated. Under the optimized conditions, the calculated detection of limit (LOD, 90% inhibition) reached 0.86 ng/mL with a wide linear range of 2.31–66.84 ng/mL, which was approximately eleven times lower than that by HRP-based traditional ELISA with the same antibody. Meanwhile, it could improve the inherent built-in rectification to the environment by the combination of the dual-output ratiometric fluorescence assays with ELISA, which also enhanced the accuracy and precision (recoveries, 87.20–106.62%; CV, 2.57–6.54%), indicating it can be applied to investigate the concentration of DBP in water samples. Full article

(This article belongs to the Special Issue Bioassays and Biosensors for Rapid Detection and Analysis)

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15 pages, 2521 KiB

Open AccessArticle

Bacterial Lighthouses—Real-Time Detection of Yersinia enterocolitica by Quorum Sensing

by Julia Niehues, Christopher McElroy, Alexander Croon, Jan Pietschmann, Martin Frettlöh and Florian Schröper

Biosensors 2021, 11(12), 517; https://doi.org/10.3390/bios11120517 - 16 Dec 2021

Cited by 1 | Viewed by 3237

Abstract

Foodborne zoonotic pathogens have a severe impact on food safety. The demand for animal-based food products (meat, milk, and eggs) is increasing, and therefore faster methods are necessary to detect infected animals or contaminated food before products enter the market. However, conventional detection is based on time-consuming microbial cultivation methods. Here, the establishment of a quorum sensing-based method for detection of foodborne pathogens as Yersinia enterocolitica in a co-cultivation approach using a bacterial biosensor carrying a special sensor plasmid is described. We combined selective enrichment with the simultaneous detection of pathogens by recording autoinducer-1-induced bioluminescent response of the biosensor. This new approach enables real-time detection with a calculated sensitivity of one initial cell in a sample after 15.3 h of co-cultivation, while higher levels of initial contamination can be detected within less than half of the time. Our new method is substantially faster than conventional microbial cultivation and should be transferrable to other zoonotic foodborne pathogens. As we could demonstrate, quorum sensing is a promising platform for the development of sensitive assays in the area of food quality, safety, and hygiene. Full article

(This article belongs to the Special Issue Bioassays and Biosensors for Rapid Detection and Analysis)

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Review

Jump to: Editorial, Research

17 pages, 3571 KiB

Open AccessReview

Strategies for Enhancing the Sensitivity of Electrochemiluminescence Biosensors

by Yueyue Huang, Yuanyuan Yao, Yueliang Wang, Lifen Chen, Yanbo Zeng, Lei Li and Longhua Guo

Biosensors 2022, 12(9), 750; https://doi.org/10.3390/bios12090750 - 11 Sep 2022

Cited by 12 | Viewed by 2798

Abstract

Electrochemiluminescence (ECL) has received considerable attention as a powerful analytical technique for the sensitive and accurate detection of biological analytes owing to its high sensitivity and selectivity and wide dynamic range. To satisfy the growing demand for ultrasensitive analysis techniques with high efficiency and accuracy in complex real sample matrices, considerable efforts have been dedicated to developing ECL strategies to improve the sensitivity of bioanalysis. As one of the most effective approaches, diverse signal amplification strategies have been integrated with ECL biosensors to achieve desirable analytical performance. This review summarizes the recent advances in ECL biosensing based on various signal amplification strategies, including DNA-assisted amplification strategies, efficient ECL luminophores, surface-enhanced electrochemiluminescence, and ratiometric strategies. Sensitivity-enhancing strategies and bio-related applications are discussed in detail. Moreover, the future trends and challenges of ECL biosensors are discussed. Full article

(This article belongs to the Special Issue Bioassays and Biosensors for Rapid Detection and Analysis)

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