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Novel Electrochemical Biosensors for Clinical Assays

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A special issue of Sensors (ISSN 1424-8220). This special issue belongs to the section "Biosensors".

Deadline for manuscript submissions: closed (31 March 2021) | Viewed by 52918

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Special Issue Editor

Prof. Dr. Antonio Guerrieri

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Dipartimento di Scienze, Università degli Studi della Basilicata, Potenza, Italy
Interests: amperometric biosensors; protein/ biomaterial interactions; enhanced sensing devices; enzyme immobilization; electrosynthesis of thin polymeric films; HPLC and FIA detection

Special Issue Information

Dear Colleagues,

Biosensors, i.e., devices where biological molecules or bio(mimetic)structures are intimately coupled to a chemo/physical transducer for converting a biorecognition event into a measurable signal, have recently gained a wide (if not huge) academic and practical interest for the multitude of their applications in analysis, especially in the field of bioanalysis, medical diagnostics, and clinical assays. Indeed, thanks to their very simple use (permitting sometimes their application at home), the minimal sample pretreatment requirement, the higher selectivity, and sensitivity, biosensors are an essential tool in the detection and monitoring of a wide range of medical conditions from glycemia to Alzheimer’s disease as well as in the monitoring of drug responses. Soon, we expect that their importance and use in clinical diagnostics will expand rapidly so as to be of critical importance to public health in the coming years.

In this context, electrochemical biosensors definitely play an innovative and quite promising role, particularly due to their clear advantages over, e.g., the spectroscopic methods, since they can be used even when the clinical samples are turbid or coloured. More important, electrochemical biosensors and the relevant instrumentations are cheap, easy to use, and usable in field analysis with a minimum or no sample pretreatment. Of course, to be effective in clinical assays, the biorecognition event of the desired analyte needs to be developed and optimized so further efforts are required in choosing/designing the required biomolecule/structure and its efficient coupling to the transducer; not less important, the flow of the bio/chemical information starting from the biorecognition event to the chemo/physical detection needs to be studied and optimized as well. On the other hand, electrochemical detection is not without its drawbacks since it is barely selective, so novel and more effective electrochemical detection approaches need to be studied and developed to assure an interferent-free detection of analytes in clinical samples without sample pretreatment.

This Special Issue focuses on research and development in the field of biosensors as analytical tools for clinical assays and medical diagnostics. It is a pleasure for me to invite you to participate to this Special Issue by submitting both original research papers and review articles.

Prof. Dr. Antonio Guerrieri
Guest Editor

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Keywords

Electrochemical biosensors
Immobilized bioreceptors
Biorecognition kinetics
Modified electrodes
Polymeric film
Molecular imprinted polymer
Nanomaterial
Sample interference
Clinical analysis
Medical diagnostics

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Published Papers (12 papers)

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12 pages, 3598 KiB

Open AccessArticle

Rapid and Sensitive Detection of miRNA Based on AC Electrokinetic Capacitive Sensing for Point-of-Care Applications

by Nan Wan, Yu Jiang, Jiamei Huang, Rania Oueslati, Shigetoshi Eda, Jayne Wu and Xiaogang Lin

Sensors 2021, 21(12), 3985; https://doi.org/10.3390/s21123985 - 9 Jun 2021

Cited by 7 | Viewed by 2499

Abstract

A sensitive and efficient method for microRNAs (miRNAs) detection is strongly desired by clinicians and, in recent years, the search for such a method has drawn much attention. There has been significant interest in using miRNA as biomarkers for multiple diseases and conditions in clinical diagnostics. Presently, most miRNA detection methods suffer from drawbacks, e.g., low sensitivity, long assay time, expensive equipment, trained personnel, or unsuitability for point-of-care. New methodologies are needed to overcome these limitations to allow rapid, sensitive, low-cost, easy-to-use, and portable methods for miRNA detection at the point of care. In this work, to overcome these shortcomings, we integrated capacitive sensing and alternating current electrokinetic effects to detect specific miRNA-16b molecules, as a model, with the limit of detection reaching 1.0 femto molar (fM) levels. The specificity of the sensor was verified by testing miRNA-25, which has the same length as miRNA-16b. The sensor we developed demonstrated significant improvements in sensitivity, response time and cost over other miRNA detection methods, and has application potential at point-of-care. Full article

(This article belongs to the Special Issue Novel Electrochemical Biosensors for Clinical Assays)

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22 pages, 3144 KiB

Open AccessArticle

A Crosstalk- and Interferent-Free Dual Electrode Amperometric Biosensor for the Simultaneous Determination of Choline and Phosphocholine

by Rosanna Ciriello and Antonio Guerrieri

Sensors 2021, 21(10), 3545; https://doi.org/10.3390/s21103545 - 19 May 2021

Cited by 2 | Viewed by 2528

Abstract

Choline (Ch) and phosphocholine (PCh) levels in tissues are associated to tissue growth and so to carcinogenesis. Till now, only highly sophisticated and expensive techniques like those based on NMR spectroscopy or GC/LC- high resolution mass spectrometry permitted Ch and PCh analysis but very few of them were capable of a simultaneous determination of these analytes. Thus, a never reported before amperometric biosensor for PCh analysis based on choline oxidase and alkaline phosphatase co-immobilized onto a Pt electrode by co-crosslinking has been developed. Coupling the developed biosensor with a parallel sensor but specific to Ch, a crosstalk-free dual electrode biosensor was also developed, permitting the simultaneous determination of Ch and PCh in flow injection analysis. This novel sensing device performed remarkably in terms of sensitivity, linear range, and limit of detection so to exceed in most cases the more complex analytical instrumentations. Further, electrode modification by overoxidized polypyrrole permitted the development of a fouling- and interferent-free dual electrode biosensor which appeared promising for the simultaneous determination of Ch and PCh in a real sample. Full article

(This article belongs to the Special Issue Novel Electrochemical Biosensors for Clinical Assays)

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13 pages, 1820 KiB

Open AccessArticle

Simple and Cost-Effective Electrochemical Method for Norepinephrine Determination Based on Carbon Dots and Tyrosinase

by Sylwia Baluta, Anna Lesiak and Joanna Cabaj

Sensors 2020, 20(16), 4567; https://doi.org/10.3390/s20164567 - 14 Aug 2020

Cited by 30 | Viewed by 4414

Abstract

Although neurotransmitters are present in human serum at the nM level, any dysfunction of the catecholamines concentration may lead to numerous serious health problems. Due to this fact, rapid and sensitive catecholamines detection is extremely important in modern medicine. However, there is no device that would measure the concentration of these compounds in body fluids. The main goal of the present study is to design a simple as possible, cost-effective new biosensor-based system for the detection of neurotransmitters, using nontoxic reagents. The miniature Au-E biosensor was designed and constructed through the immobilization of tyrosinase on an electroactive layer of cysteamine and carbon nanoparticles covering the gold electrode. This sensing arrangement utilized the catalytic oxidation of norepinephrine (NE) to NE quinone, measured with voltammetric techniques: cyclic voltammetry and differential pulse voltammetry. The prepared bio-system exhibited good parameters: a broad linear range (1–200 μM), limit of detection equal to 196 nM, limit of quantification equal to 312 nM, and high selectivity and sensitivity. It is noteworthy that described method was successfully applied for NE determination in real samples. Full article

(This article belongs to the Special Issue Novel Electrochemical Biosensors for Clinical Assays)

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12 pages, 3299 KiB

Open AccessArticle

In Vivo Optogenetic Modulation with Simultaneous Neural Detection Using Microelectrode Array Integrated with Optical Fiber

by Penghui Fan, Yilin Song, Shengwei Xu, Yuchuan Dai, Yiding Wang, Botao Lu, Jingyu Xie, Hao Wang and Xinxia Cai

Sensors 2020, 20(16), 4526; https://doi.org/10.3390/s20164526 - 13 Aug 2020

Cited by 5 | Viewed by 3076

Abstract

The detection of neuroelectrophysiology while performing optogenetic modulation can provide more reliable and useful information for neural research. In this study, an optical fiber and a microelectrode array were integrated through hot-melt adhesive bonding, which combined optogenetics and electrophysiological detection technology to achieve neuromodulation and neuronal activity recording. We carried out the experiments on the activation and electrophysiological detection of infected neurons at the depth range of 900–1250 μm in the brain which covers hippocampal CA1 and a part of the upper cortical area, analyzed a possible local inhibition circuit by combining opotogenetic modulation and electrophysiological characteristics and explored the effects of different optical patterns and light powers on the neuromodulation. It was found that optogenetics, combined with neural recording technology, could provide more information and ideas for neural circuit recognition. In this study, the optical stimulation with low frequency and large duty cycle induces more intense neuronal activity and larger light power induced more action potentials of neurons within a certain power range (1.032 mW–1.584 mW). The present study provided an efficient method for the detection and modulation of neurons in vivo and an effective tool to study neural circuit in the brain. Full article

(This article belongs to the Special Issue Novel Electrochemical Biosensors for Clinical Assays)

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10 pages, 929 KiB

Open AccessCommunication

Application of a Glucose Dehydrogenase-Fused with Zinc Finger Protein to Label DNA Aptamers for the Electrochemical Detection of VEGF

by Jinhee Lee, Atsuro Tatsumi, Kaori Tsukakoshi, Ellie D. Wilson, Koichi Abe, Koji Sode and Kazunori Ikebukuro

Sensors 2020, 20(14), 3878; https://doi.org/10.3390/s20143878 - 11 Jul 2020

Cited by 13 | Viewed by 3124

Abstract

Aptamer-based electrochemical sensors have gained attention in the context of developing a diagnostic biomarker detection method because of their rapid response, miniaturization ability, stability, and design flexibility. In such detection systems, enzymes are often used as labels to amplify the electrochemical signal. We have focused on glucose dehydrogenase (GDH) as a labeling enzyme for electrochemical detection owing to its high enzymatic activity, availability, and well-established electrochemical principle and platform. However, it is difficult and laborious to obtain one to one labeling of a GDH-aptamer complex with conventional chemical conjugation methods. In this study, we used GDH that was genetically fused to a DNA binding protein, i.e., zinc finger protein (ZF). Fused GDH can be attached to an aptamer spontaneously and site specifically in a buffer by exploiting the sequence-specific binding ability of ZF. Using such a fusion protein, we labeled a vascular endothelial growth factor (VEGF)-binding aptamer with GDH and detected the target electrochemically. As a result, upon the addition of glucose, the GDH labeled on the aptamer generated an amperometric signal, and the current response increased dependent on the VEGF concentration. Eventually, the developed electrochemical sensor proved to detect VEGF levels as low as 105 pM, thereby successfully demonstrating the concept of using ZF-fused GDH to enzymatically label aptamers. Full article

(This article belongs to the Special Issue Novel Electrochemical Biosensors for Clinical Assays)

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14 pages, 2112 KiB

Open AccessArticle

Employment of 1-Methoxy-5-Ethyl Phenazinium Ethyl Sulfate as a Stable Electron Mediator in Flavin Oxidoreductases-Based Sensors

by Maya Fitriana, Noya Loew, Arief Budi Witarto, Kazunori Ikebukuro, Koji Sode and Wakako Tsugawa

Sensors 2020, 20(10), 2825; https://doi.org/10.3390/s20102825 - 15 May 2020

Cited by 7 | Viewed by 4127

Abstract

In this paper, a novel electron mediator, 1-methoxy-5-ethyl phenazinium ethyl sulfate (mPES), was introduced as a versatile mediator for disposable enzyme sensor strips, employing representative flavin oxidoreductases, lactate oxidase (LOx), glucose dehydrogenase (GDH), and fructosyl peptide oxidase (FPOx). A disposable lactate enzyme sensor with oxygen insensitive Aerococcus viridans-derived engineered LOx (AvLOx), with A96L mutant as the enzyme, was constructed. The constructed lactate sensor exhibited a high sensitivity (0.73 ± 0.12 μA/mM) and wide linear range (0–50 mM lactate), showings that mPES functions as an effective mediator for AvLOx. Employing mPES as mediator allowed this amperometric lactate sensor to be operated at a relatively low potential of +0.2 V to 0 V vs. Ag/AgCl, thus avoiding interference from uric acid and acetaminophen. The lactate sensors were adequately stable for at least 48 days of storage at 25 °C. These results indicated that mPES can be replaced with 1-methoxy-5-methyl phenazinium methyl sulfate (mPMS), which we previously reported as the best mediator for AvLOx-based lactate sensors. Furthermore, this study revealed that mPES can be used as an effective electron mediator for the enzyme sensors employing representative flavin oxidoreductases, GDH-based glucose sensors, and FPOx-based hemoglobin A1c (HbA1c) sensors. Full article

(This article belongs to the Special Issue Novel Electrochemical Biosensors for Clinical Assays)

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12 pages, 1973 KiB

Open AccessArticle

A Self-Calibrating IoT Portable Electrochemical Immunosensor for Serum Human Epididymis Protein 4 as a Tumor Biomarker for Ovarian Cancer

by Valentina Bianchi, Monica Mattarozzi, Marco Giannetto, Andrea Boni, Ilaria De Munari and Maria Careri

Sensors 2020, 20(7), 2016; https://doi.org/10.3390/s20072016 - 3 Apr 2020

Cited by 23 | Viewed by 3847

Abstract

Nowadays, analytical techniques are moving towards the development of smart biosensing strategies for the point-of-care accurate screening of disease biomarkers, such as human epididymis protein 4 (HE4), a recently discovered serum marker for early ovarian cancer diagnosis. In this context, the present work represents the first implementation of a competitive enzyme-labelled magneto-immunoassay exploiting a homemade IoT Wi-Fi cloud-based portable potentiostat for differential pulse voltammetry readout. The electrochemical device was specifically designed to be capable of autonomous calibration and data processing, switching between calibration, and measurement modes: in particular, firstly, a baseline estimation algorithm is applied for correct peak computation, then calibration function is built by interpolating data with a four-parameter logistic function. The calibration function parameters are stored on the cloud for inverse prediction to determine the concentration of unknown samples. Interpolation function calibration and concentration evaluation are performed directly on-board, thus reducing the power consumption. The analytical device was validated in human serum, demonstrating good sensing performance for analysis of HE4 with detection and quantitation limits in human serum of 3.5 and 29.2 pM, respectively, reaching the sensitivity that is required for diagnostic purposes, with high potential for applications as portable and smart diagnostic tool for point-of-care testing. Full article

(This article belongs to the Special Issue Novel Electrochemical Biosensors for Clinical Assays)

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17 pages, 2473 KiB

Open AccessArticle

Assay of Phospholipase D Activity by an Amperometric Choline Oxidase Biosensor

by Rosanna Ciriello and Antonio Guerrieri

Sensors 2020, 20(5), 1304; https://doi.org/10.3390/s20051304 - 27 Feb 2020

Cited by 5 | Viewed by 3025

Abstract

A novel electrochemical method to assay phospholipase D (PLD) activity is proposed based on the employment of a choline biosensor realized by immobilizing choline oxidase through co-crosslinking on an overoxidized polypyrrole film previously deposited on a platinum electrode. To perform the assay, an aliquot of a PLD standard solution is typically added to borate buffer containing phosphatidylcholine at a certain concentration and the oxidation current of hydrogen peroxide is then measured at the rotating modified electrode by applying a detection potential of +0.7 V vs. SCE. Various experimental parameters influencing the assay were studied and optimized. The employment of 0.75% (v/v) Triton X-100, 0.2 mM calcium chloride, 5 mM phosphatidylcholine, and borate buffer at pH 8.0, ionic strength (I) 0.05 M allowed to achieve considerable current responses. In order to assure a controlled mass transport and, at the same time, high sensitivity, an electrode rotation rate of 200 rpm was selected. The proposed method showed a sensitivity of 24 (nA/s)⋅(IU/mL)⁻¹, a wide linear range up to 0.33 IU/mL, fast response time and appreciable long-term stability. The limit of detection, evaluated from the linear calibration curve, was 0.005 IU/mL (S/N = 3). Finally, due to the presence of overoxidized polypyrrole film characterized by notable rejection properties towards electroactive compounds, a practical application to real sample analysis can be envisaged. Full article

(This article belongs to the Special Issue Novel Electrochemical Biosensors for Clinical Assays)

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Review

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15 pages, 2787 KiB

Open AccessReview

Determination of Folic Acid Using Biosensors—A Short Review of Recent Progress

by Alessio Di Tinno, Rocco Cancelliere and Laura Micheli

Sensors 2021, 21(10), 3360; https://doi.org/10.3390/s21103360 - 12 May 2021

Cited by 16 | Viewed by 3869

Abstract

Folic acid (FA) is the synthetic surrogate of the essential B vitamin folate, alternatively named folacin, pteroylglutamic acid or vitamin B₉. FA is an electroactive compound that helps our body to create and keep our cells healthy: it acts as the main character in a variety of synthetic biological reactions such as the synthesis of purines, pyrimidine (thus being indirectly implied in DNA synthesis), fixing and methylation of DNA. Therefore, physiological folate deficiency may be responsible for severe degenerative conditions, including neural tube defects in developing embryos and megaloblastic anaemia at any age. Moreover, being a water-soluble molecule, it is constantly lost and has to be reintegrated daily; for this reason, FA supplements and food fortification are, nowadays, extremely diffused and well-established practices. Consequently, accurate, reliable and precise analytical techniques are needed to exactly determine FA concentration in various media. Thus, the aim of this review is to report on research papers of the past 5 years (2016–2020) dealing with rapid and low-cost electrochemical determination of FA in food or biological fluid samples. Full article

(This article belongs to the Special Issue Novel Electrochemical Biosensors for Clinical Assays)

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37 pages, 16224 KiB

Open AccessReview

Electrochemical Nanobiosensors for Detection of Breast Cancer Biomarkers

by Veronika Gajdosova, Lenka Lorencova, Peter Kasak and Jan Tkac

Sensors 2020, 20(14), 4022; https://doi.org/10.3390/s20144022 - 20 Jul 2020

Cited by 45 | Viewed by 8292

Abstract

This comprehensive review paper describes recent advances made in the field of electrochemical nanobiosensors for the detection of breast cancer (BC) biomarkers such as specific genes, microRNA, proteins, circulating tumor cells, BC cell lines, and exosomes or exosome-derived biomarkers. Besides the description of key functional characteristics of electrochemical nanobiosensors, the reader can find basic statistic information about BC incidence and mortality, breast pathology, and current clinically used BC biomarkers. The final part of the review is focused on challenges that need to be addressed in order to apply electrochemical nanobiosensors in a clinical practice. Full article

(This article belongs to the Special Issue Novel Electrochemical Biosensors for Clinical Assays)

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14 pages, 2584 KiB

Open AccessReview

Molecularly Imprinted Polymers and Surface Imprinted Polymers Based Electrochemical Biosensor for Infectious Diseases

by Feiyun Cui, Zhiru Zhou and H. Susan Zhou

Sensors 2020, 20(4), 996; https://doi.org/10.3390/s20040996 - 13 Feb 2020

Cited by 147 | Viewed by 9921

Abstract

Owing to their merits of simple, fast, sensitive, and low cost, electrochemical biosensors have been widely used for the diagnosis of infectious diseases. As a critical element, the receptor determines the selectivity, stability, and accuracy of the electrochemical biosensors. Molecularly imprinted polymers (MIPs) and surface imprinted polymers (SIPs) have great potential to be robust artificial receptors. Therefore, extensive studies have been reported to develop MIPs/SIPs for the detection of infectious diseases with high selectivity and reliability. In this review, we discuss mechanisms of recognition events between imprinted polymers with different biomarkers, such as signaling molecules, microbial toxins, viruses, and bacterial and fungal cells. Then, various preparation methods of MIPs/SIPs for electrochemical biosensors are summarized. Especially, the methods of electropolymerization and micro-contact imprinting are emphasized. Furthermore, applications of MIPs/SIPs based electrochemical biosensors for infectious disease detection are highlighted. At last, challenges and perspectives are discussed. Full article

(This article belongs to the Special Issue Novel Electrochemical Biosensors for Clinical Assays)

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Other

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11 pages, 2286 KiB

Open AccessLetter

Assessment of Gold Bio-Functionalization for Wide-Interface Biosensing Platforms

by Lucia Sarcina, Luisa Torsi, Rosaria Anna Picca, Kyriaki Manoli and Eleonora Macchia

Sensors 2020, 20(13), 3678; https://doi.org/10.3390/s20133678 - 30 Jun 2020

Cited by 14 | Viewed by 3173

Abstract

The continuous improvement of the technical potential of bioelectronic devices for biosensing applications will provide clinicians with a reliable tool for biomarker quantification down to the single molecule. Eventually, physicians will be able to identify the very moment at which the illness state begins, with a terrific impact on the quality of life along with a reduction of health care expenses. However, in clinical practice, to gather enough information to formulate a diagnosis, multiple biomarkers are normally quantified from the same biological sample simultaneously. Therefore, it is critically important to translate lab-based bioelectronic devices based on electrolyte gated thin-film transistor technology into a cost-effective portable multiplexing array prototype. In this perspective, the assessment of cost-effective manufacturability represents a crucial step, with specific regard to the optimization of the bio-functionalization protocol of the transistor gate module. Hence, we have assessed, using surface plasmon resonance technique, a sustainable and reliable cost-effective process to successfully bio-functionalize a gold surface, suitable as gate electrode for wide-field bioelectronic sensors. The bio-functionalization process herein investigated allows to reduce the biorecognition element concentration to one-tenth, drastically impacting the manufacturing costs while retaining high analytical performance. Full article

(This article belongs to the Special Issue Novel Electrochemical Biosensors for Clinical Assays)

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