Electrochemical Immunosensors and Aptasensors

A special issue of Chemosensors (ISSN 2227-9040).

Deadline for manuscript submissions: closed (31 August 2016) | Viewed by 94381

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Department of Molecular Sciences and Nanosystems, University Ca’ Foscari of Venice, via Torino 155, 30172 Venezia Mestre, Italy
Interests: molecular electrochemistry; electrochemosensors and biosensors; environmental electroanalysis; nanoelectrodes and bio-nanoelectrochemistry
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Department of Molecular Sciences and Nanosystems, University Ca' Foscari of Venice, Via Torino 155, 30172 Mestre, Italy
Interests: development of electrochemical sensors and biosensors for electrochemical and biomedical applications; environmental electroanalysis; modified electrodes; nanoelectrodes and arrays of nanoelectrodes; nanostructured electrodes
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Special Issue Information

Dear Colleagues,

The possibility to integrate biorecognition elements into electrochemical detection systems has opened the way to a new class of powerful analytical devices named electrochemical (EC) biosensors. The first EC biosensors have employed enzymes as recognition element, however this has limited their application to redox enzymes and their natural or artificial redox substrates or inhibitors. The widening towards non-electroactive analytes was later possible thanks to the development of affinity sensors in which specific interactions between biomolecules are exploited for developing highly selective and sensitive biosensors. Presently, the combination of the exceptional molecular recognition capabilities of antibodies and aptamers with the sensitivity, low cost, practicality of use and handiness of electrochemical devices is leading to an impressive development of EC immunosensors and aptasensors, potentially suitable to detect a wide range of analytes, mainly (but not only) of a proteic nature, following a path that is moving side by side with the most recent advances in proteomics. Interestingly, together with continuous improvements and refinements in EC immunosensors based on the use of labels, intrinsically electroactive or able to interact with electroactive molecules, a new generation of label-free sensors is being developed.

The aim of this Special Issue of Chemosensors is take stock of the state of the art and identify prospects for EC immuno- and aptasensors, both labeled and label-free. Emphasis will be placed on analytical applications for the rapid detection of markers of diseases (e.g., cancer or heart attack), for toxicological and environmental control, for food safety and cultural heritage diagnostics.

Prof. Dr. Paolo Ugo
Prof. Dr. Ligia Moretto
Guest Editors

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Keywords

  • Electrochemical sensors
  • Biosensors
  • Immuno-test
  • Aptamers
  • Protein detection
  • Voltammetry
  • Amperometry
  • Electrochemical impedance
  • Functional electrodes

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Published Papers (12 papers)

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Editorial

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155 KiB  
Editorial
Electrochemical Immunosensors and Aptasensors
by Paolo Ugo and Ligia M. Moretto
Chemosensors 2017, 5(2), 13; https://doi.org/10.3390/chemosensors5020013 - 2 Apr 2017
Cited by 11 | Viewed by 4746
Abstract
Since the first electrochemical biosensor for glucose detection, pioneered in 1962 by Clark and Lyons [1], research and application in the field has grown at an impressive rate and we are still witnessing a continuing evolution of research on this topic [2].[...] Full article
(This article belongs to the Special Issue Electrochemical Immunosensors and Aptasensors)

Research

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2820 KiB  
Article
Electrochemical Immunosensor for Detection of IgY in Food and Food Supplements
by Chiara Gaetani, Emmanuele Ambrosi, Paolo Ugo and Ligia M. Moretto
Chemosensors 2017, 5(1), 10; https://doi.org/10.3390/chemosensors5010010 - 2 Mar 2017
Cited by 14 | Viewed by 6029
Abstract
Immunoglobulin Y is a water-soluble protein present in high concentration in hen serum and egg yolk. IgY has applications in many fields, e.g., from food stuff to the mass production of antibodies. In this work, we have implemented an electrochemical immunosensor for IgY [...] Read more.
Immunoglobulin Y is a water-soluble protein present in high concentration in hen serum and egg yolk. IgY has applications in many fields, e.g., from food stuff to the mass production of antibodies. In this work, we have implemented an electrochemical immunosensor for IgY based on templated nanoelectrodes ensembles. IgY is captured by the templating polycarbonate and reacted with anti-IgY labeled with horseradish peroxidase. In the presence of H2O2 and methylene blue as the redox mediator, an electrocatalytic current is generated which scales with IgY concentration in the sample. After optimizing the extracting procedure, the immunosensor was applied for analysis of fresh eggs and food integrators. The data obtained with the biosensor were validated by SDS-PAGE and Western blot measurements. Full article
(This article belongs to the Special Issue Electrochemical Immunosensors and Aptasensors)
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942 KiB  
Article
Design of an Affibody-Based Recognition Strategy for Human Epidermal Growth Factor Receptor 2 (HER2) Detection by Electrochemical Biosensors
by Hoda Ilkhani, Andrea Ravalli and Giovanna Marrazza
Chemosensors 2016, 4(4), 23; https://doi.org/10.3390/chemosensors4040023 - 2 Dec 2016
Cited by 21 | Viewed by 5884
Abstract
In this study, we have designed and realized three simple electrochemical bioassays for the detection of the human epidermal growth factor receptor 2 (HER2) cancer biomarker using magnetic beads coupling screen-printed arrays. The different approaches were based on a sandwich format in which [...] Read more.
In this study, we have designed and realized three simple electrochemical bioassays for the detection of the human epidermal growth factor receptor 2 (HER2) cancer biomarker using magnetic beads coupling screen-printed arrays. The different approaches were based on a sandwich format in which affibody (Af) or antibody (Ab) molecules were coupled respectively to streptavidin or protein A-modified magnetic beads. The bioreceptor-modified beads were used to capture the HER2 protein from the sample and sandwich assay was performed by adding the labeled secondary affibody or the antibody. An enzyme-amplified detection scheme based on the coupling of secondary biotinylated bioreceptor with streptavidin-alkaline phosphatase enzyme conjugate was then applied. The enzyme catalyzed the hydrolysis of the electro-inactive 1-naphthyl-phosphate to the electro-active 1-naphthol, which was detected by means of differential pulse voltammetry (DPV). Each developed assay has been studied and optimized. Furthermore, a thorough comparison of the analytical performances of developed assays was performed. Finally, preliminary experiments using serum samples spiked with HER2 protein were also carried out. Full article
(This article belongs to the Special Issue Electrochemical Immunosensors and Aptasensors)
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3255 KiB  
Article
Miniaturized Aptamer-Based Assays for Protein Detection
by Alessandro Bosco, Elena Ambrosetti, Jan Mavri, Pietro Capaldo and Loredana Casalis
Chemosensors 2016, 4(3), 18; https://doi.org/10.3390/chemosensors4030018 - 2 Sep 2016
Cited by 6 | Viewed by 6914
Abstract
The availability of devices for cancer biomarker detection at early stages of the disease is one of the most critical issues in biomedicine. Towards this goal, to increase the assay sensitivity, device miniaturization strategies empowered by the employment of high affinity protein binders [...] Read more.
The availability of devices for cancer biomarker detection at early stages of the disease is one of the most critical issues in biomedicine. Towards this goal, to increase the assay sensitivity, device miniaturization strategies empowered by the employment of high affinity protein binders constitute a valuable approach. In this work we propose two different surface-based miniaturized platforms for biomarker detection in body fluids: the first platform is an atomic force microscopy (AFM)-based nanoarray, where AFM is used to generate functional nanoscale areas and to detect biorecognition through careful topographic measurements; the second platform consists of a miniaturized electrochemical cell to detect biomarkers through electrochemical impedance spectroscopy (EIS) analysis. Both devices rely on robust and highly-specific protein binders as aptamers, and were tested for thrombin detection. An active layer of DNA-aptamer conjugates was immobilized via DNA directed immobilization on complementary single-stranded DNA self-assembled monolayers confined on a nano/micro area of a gold surface. Results obtained with these devices were compared with the output of surface plasmon resonance (SPR) assays used as reference. We succeeded in capturing antigens in concentrations as low as a few nM. We put forward ideas to push the sensitivity further to the pM range, assuring low biosample volume (μL range) assay conditions. Full article
(This article belongs to the Special Issue Electrochemical Immunosensors and Aptasensors)
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1465 KiB  
Article
Unique Properties of Core Shell Ag@Au Nanoparticles for the Aptasensing of Bacterial Cells
by Ezat Hamidi-Asl, Freddy Dardenne, Sanaz Pilehvar, Ronny Blust and Karolien De Wael
Chemosensors 2016, 4(3), 16; https://doi.org/10.3390/chemosensors4030016 - 29 Aug 2016
Cited by 39 | Viewed by 7861
Abstract
In this article, it is shown that the efficiency of an electrochemical aptasensing device is influenced by the use of different nanoparticles (NPs) such as gold nanoparticles (Au), silver nanoparticles (Ag), hollow gold nanospheres (HGN), hollow silver nanospheres (HSN), silver–gold core shell (Ag@Au), [...] Read more.
In this article, it is shown that the efficiency of an electrochemical aptasensing device is influenced by the use of different nanoparticles (NPs) such as gold nanoparticles (Au), silver nanoparticles (Ag), hollow gold nanospheres (HGN), hollow silver nanospheres (HSN), silver–gold core shell (Ag@Au), gold–silver core shell (Au@Ag), and silver–gold alloy nanoparticles (Ag/Au). Among these nanomaterials, Ag@Au core shell NPs are advantageous for aptasensing applications because the core improves the physical properties and the shell provides chemical stability and biocompatibility for the immobilization of aptamers. Self-assembly of the NPs on a cysteamine film at the surface of a carbon paste electrode is followed by the immobilization of thiolated aptamers at these nanoframes. The nanostructured (Ag@Au) aptadevice for Escherichia coli as a target shows four times better performance in comparison to the response obtained at an aptamer modified planar gold electrode. A comparison with other (core shell) NPs is performed by cyclic voltammetry and differential pulse voltammetry. Also, the selectivity of the aptasensor is investigated using other kinds of bacteria. The synthesized NPs and the morphology of the modified electrode are characterized by UV-Vis absorption spectroscopy, scanning electron microscopy, energy dispersive X-ray analysis, and electrochemical impedance spectroscopy. Full article
(This article belongs to the Special Issue Electrochemical Immunosensors and Aptasensors)
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972 KiB  
Article
Simultaneous Determination of the Main Peanut Allergens in Foods Using Disposable Amperometric Magnetic Beads-Based Immunosensing Platforms
by Víctor Ruiz-Valdepeñas Montiel, Rebeca Magnolia Torrente-Rodríguez, Susana Campuzano, Alessandro Pellicanò, Ángel Julio Reviejo, Maria Stella Cosio and José Manuel Pingarrón
Chemosensors 2016, 4(3), 11; https://doi.org/10.3390/chemosensors4030011 - 28 Jun 2016
Cited by 19 | Viewed by 7859
Abstract
In this work, a novel magnetic beads (MBs)-based immunosensing approach for the rapid and simultaneous determination of the main peanut allergenic proteins (Ara h 1 and Ara h 2) is reported. It involves the use of sandwich-type immunoassays using selective capture and detector [...] Read more.
In this work, a novel magnetic beads (MBs)-based immunosensing approach for the rapid and simultaneous determination of the main peanut allergenic proteins (Ara h 1 and Ara h 2) is reported. It involves the use of sandwich-type immunoassays using selective capture and detector antibodies and carboxylic acid-modified magnetic beads (HOOC-MBs). Amperometric detection at −0.20 V was performed using dual screen-printed carbon electrodes (SPdCEs) and the H2O2/hydroquinone (HQ) system. This methodology exhibits high sensitivity and selectivity for the target proteins providing detection limits of 18.0 and 0.07 ng/mL for Ara h 1 and Ara h 2, respectively, with an assay time of only 2 h. The usefulness of the approach was evaluated by detecting the endogenous content of both allergenic proteins in different food extracts as well as trace amounts of peanut allergen (0.0001% or 1.0 mg/kg) in wheat flour spiked samples. The developed platform provides better Low detection limits (LODs) in shorter assay times than those claimed for the allergen specific commercial ELISA kits using the same immunoreagents and quantitative information on individual food allergen levels. Moreover, the flexibility of the methodology makes it readily translate to the detection of other food-allergens. Full article
(This article belongs to the Special Issue Electrochemical Immunosensors and Aptasensors)
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Review

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2112 KiB  
Review
Recent Advances in Electrochemical-Based Sensing Platforms for Aflatoxins Detection
by Atul Sharma, Kotagiri Yugender Goud, Akhtar Hayat, Sunil Bhand and Jean Louis Marty
Chemosensors 2017, 5(1), 1; https://doi.org/10.3390/chemosensors5010001 - 26 Dec 2016
Cited by 38 | Viewed by 8343
Abstract
Mycotoxin are small (MW ~700 Da), toxic secondary metabolites produced by fungal species that readily colonize crops and contaminate them at both pre- and post-harvesting. Among all, aflatoxins (AFs) are mycotoxins of major significance due to their presence in common food commodities and [...] Read more.
Mycotoxin are small (MW ~700 Da), toxic secondary metabolites produced by fungal species that readily colonize crops and contaminate them at both pre- and post-harvesting. Among all, aflatoxins (AFs) are mycotoxins of major significance due to their presence in common food commodities and the potential threat to human health worldwide. Based on the severity of illness and increased incidences of AFs poisoning, a broad range of conventional and analytical detection techniques that could be useful and practical have already been reported. However, due to the variety of structural analogous of these toxins, it is impossible to use one common technique for their analysis. Numerous recent research efforts have been directed to explore alternative detection technologies. Recently, immunosensors and aptasensors have gained promising potential in the area of sample preparation and detection systems. These sensors offer the advantages of disposability, portability, miniaturization, and on-site analysis. In a typical design of an aptasensor, an aptamer (ssDNA or RNA) is used as a bio-recognition element either integrated within or in intimate association with the transducer surface. This review paper is focused on the recent advances in electrochemical immuno- and aptasensing platforms for detection of AFs in real samples. Full article
(This article belongs to the Special Issue Electrochemical Immunosensors and Aptasensors)
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2391 KiB  
Review
Recent Trends in Field-Effect Transistors-Based Immunosensors
by Ana Carolina Mazarin De Moraes and Lauro Tatsuo Kubota
Chemosensors 2016, 4(4), 20; https://doi.org/10.3390/chemosensors4040020 - 21 Oct 2016
Cited by 82 | Viewed by 13819
Abstract
Immunosensors are analytical platforms that detect specific antigen-antibody interactions and play an important role in a wide range of applications in biomedical clinical diagnosis, food safety, and monitoring contaminants in the environment. Field-effect transistors (FET) immunosensors have been developed as promising alternatives to [...] Read more.
Immunosensors are analytical platforms that detect specific antigen-antibody interactions and play an important role in a wide range of applications in biomedical clinical diagnosis, food safety, and monitoring contaminants in the environment. Field-effect transistors (FET) immunosensors have been developed as promising alternatives to conventional immunoassays, which require complicated processes and long-time data acquisition. The electrical signal of FET-based immunosensors is generated as a result of the antigen-antibody conjugation. FET biosensors present real-time and rapid response, require small sample volume, and exhibit higher sensitivity and selectivity. This review brings an overview on the recent literature of FET-based immunosensors, highlighting a diversity of nanomaterials modified with specific receptors as immunosensing platforms for the ultrasensitive detection of various biomolecules. Full article
(This article belongs to the Special Issue Electrochemical Immunosensors and Aptasensors)
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7261 KiB  
Review
Guanine Quadruplex Electrochemical Aptasensors
by Ana-Maria Chiorcea-Paquim and Ana Maria Oliveira-Brett
Chemosensors 2016, 4(3), 13; https://doi.org/10.3390/chemosensors4030013 - 30 Jul 2016
Cited by 15 | Viewed by 7725
Abstract
Guanine-rich nucleic acids are able to self-assemble into G-quadruplex four-stranded secondary structures, which are found at the level of telomeric regions of chromosomes, oncogene promoter sequences and other biologically-relevant regions of the genome. Due to their extraordinary stiffness and biological role, G-quadruples become [...] Read more.
Guanine-rich nucleic acids are able to self-assemble into G-quadruplex four-stranded secondary structures, which are found at the level of telomeric regions of chromosomes, oncogene promoter sequences and other biologically-relevant regions of the genome. Due to their extraordinary stiffness and biological role, G-quadruples become relevant in areas ranging from structural biology to medicinal chemistry, supra-molecular chemistry, nanotechnology and biosensor technology. In addition to classical methodologies, such as circular dichroism, nuclear magnetic resonance or crystallography, electrochemical methods have been successfully used for the rapid detection of the conformational changes from single-strand to G-quadruplex. This review presents recent advances on the G-quadruplex electrochemical characterization and on the design and applications of G-quadruplex electrochemical biosensors, with special emphasis on the G-quadruplex aptasensors and hemin/G-quadruplex peroxidase-mimicking DNAzyme biosensors. Full article
(This article belongs to the Special Issue Electrochemical Immunosensors and Aptasensors)
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4334 KiB  
Review
Aptasensors Based on Stripping Voltammetry
by Wenjing Qi, Di Wu, Guobao Xu, Jacques Nsabimana and Anaclet Nsabimana
Chemosensors 2016, 4(3), 12; https://doi.org/10.3390/chemosensors4030012 - 15 Jul 2016
Cited by 7 | Viewed by 6585
Abstract
Aptasensors based on stripping voltammetry exhibit several advantages, such as high sensitivity and multi-target detection from stripping voltammetric technology, and high selectivity from the specific binding of apamers with targets. This review comprehensively discusses the recent accomplishments in signal amplification strategies based on [...] Read more.
Aptasensors based on stripping voltammetry exhibit several advantages, such as high sensitivity and multi-target detection from stripping voltammetric technology, and high selectivity from the specific binding of apamers with targets. This review comprehensively discusses the recent accomplishments in signal amplification strategies based on nanomaterials, such as metal nanoparticles, semiconductor nanoparticles, and nanocomposite materials, which are detected by stripping voltammetry after suitable dissolution. Focus will be put in discussing multiple amplification strategies that are widely applied in aptasensors for small biomolecules, proteins, disease markers, and cancer cells. Full article
(This article belongs to the Special Issue Electrochemical Immunosensors and Aptasensors)
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1419 KiB  
Review
Aptamer-Based Electrochemical Sensing of Lysozyme
by Alina Vasilescu, Qian Wang, Musen Li, Rabah Boukherroub and Sabine Szunerits
Chemosensors 2016, 4(2), 10; https://doi.org/10.3390/chemosensors4020010 - 14 Jun 2016
Cited by 50 | Viewed by 12095
Abstract
Protein analysis and quantification are required daily by thousands of laboratories worldwide for activities ranging from protein characterization to clinical diagnostics. Multiple factors have to be considered when selecting the best detection and quantification assay, including the amount of protein available, its concentration, [...] Read more.
Protein analysis and quantification are required daily by thousands of laboratories worldwide for activities ranging from protein characterization to clinical diagnostics. Multiple factors have to be considered when selecting the best detection and quantification assay, including the amount of protein available, its concentration, the presence of interfering molecules, as well as costs and rapidity. This is also the case for lysozyme, a 14.3-kDa protein ubiquitously present in many organisms, that has been identified with a variety of functions: antibacterial activity, a biomarker of several serious medical conditions, a potential allergen in foods or a model of amyloid-type protein aggregation. Since the design of the first lysozyme aptamer in 2001, lysozyme became one of the most intensively-investigated biological target analytes for the design of novel biosensing concepts, particularly with regards to electrochemical aptasensors. In this review, we discuss the state of the art of aptamer-based electrochemical sensing of lysozyme, with emphasis on sensing in serum and real samples. Full article
(This article belongs to the Special Issue Electrochemical Immunosensors and Aptasensors)
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Other

1390 KiB  
Technical Note
A Low-Cost Label-Free AFB1 Impedimetric Immunosensor Based on Functionalized CD-Trodes
by Marcos Vinicius Foguel, Gabriela Furlan Giordano, Célia Maria De Sylos, Iracilda Zeppone Carlos, Antonio Aparecido Pupim Ferreira, Assis Vicente Benedetti and Hideko Yamanaka
Chemosensors 2016, 4(3), 17; https://doi.org/10.3390/chemosensors4030017 - 1 Sep 2016
Cited by 10 | Viewed by 5060
Abstract
This work describes the investigation of a label-free immunosensor for the detection of aflatoxin B1 (AFB1). CD-trodes (electrodes obtained from recordable compact disks) were used as low-cost and disposable transducers after modification with a self-assembled monolayer (SAM) of lipoic acid. [...] Read more.
This work describes the investigation of a label-free immunosensor for the detection of aflatoxin B1 (AFB1). CD-trodes (electrodes obtained from recordable compact disks) were used as low-cost and disposable transducers after modification with a self-assembled monolayer (SAM) of lipoic acid. The anti-aflatoxin B1 antibody was immobilized via EDC/NHS activation, followed by blocking with bovine serum albumin and immunoassays with AFB1. The optimization of analytical parameters and the detection were carried out using electrochemical impedance measurements. Using chemometric tools, the best conditions for the immunosensor development were defined as: anti-AFB1 antibody at 1:2000 dilution and surface blocking with 0.5% bovine serum albumin, both incubated for 1 h, and antibody–antigen immunoreaction for 30 min. The impedimetric immunosensor showed a linear range from 5 × 10−9 to 1 × 10−7 mol·L−1 (1.56–31.2 ng·mL−1), limit of detection and limit of quantification, respectively, 3.6 × 10−10 and 1.1 × 10−9mol·L−1 (0.11 and 0.34 ng·mL−1). The proposed immunosensor was applied to analyze peanut samples. Full article
(This article belongs to the Special Issue Electrochemical Immunosensors and Aptasensors)
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