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Special Issue "Enzymatic Biosensors"

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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 January 2013)

Special Issue Editor

Guest Editor
Dr. Jan Halámek

Department of Chemistry and Biomolecular Science, 214 Science Center, Clarkson University, Potsdam, NY 13699-5810, USA
E-Mail
Phone: 1-315-268-2354
Fax: +1 315 268 6610
Interests: enzymatic sensors; affinity sensors; bioelectronics; bionanotechnology; biosensors; bioelectrochemistry

Special Issue Information

Dear Colleagues,

The glucose biosensor, a device that changed the lives of millions affected by diabetes, is a one remarkable example of enzyme-based biosensor technology. Enzyme sensors are a major part of biosensorics - technology, which currently represents a mature analogue to instrumental analytical techniques in areas of clinical diagnostics and is the leading technology in point-of-care analysis.

Enzyme biosensors employ the affinity and selectivity of catalytically active proteins, towards their target molecules. Typically, (enzyme,usually immobilized on/within the surface of transducer - acts as a catalyst when interacting with the analyte, represented by its substrate, inhibitor, co-substrate or co-factor, while the enzyme itself remains unchanged. The transducer converts the effect created by the interaction of enzyme with the analyte, usually into an electrical signal. Depending on the assay type, two fundamental classes of enzyme sensors can be distinguished. First, the enzyme detects the presence of a substrate, or co-substrate/co-factor. This is then, by way of a transducer, used to monitor the increase of enzymatic activity. A typical example is a glucose biosensor. The second group is based on the detection of inhibitors in the presence of a substrate. With this system the decrease of signal (caused by enzyme inhibition) is monitored. The most common example of this approach is the detection of organophosphate compounds used as pesticides or warfare nerve agents. The mode of signal transduction can be electrochemical, optical, resonant (acoustic), thermal etc. The major advantage of all of these approaches is the high sensitivity and specificity of the biorecognition of a single selected analyte.

There have been significant improvements in the field of enzymatic biosensors; the usage of new, genetically engineered enzymes has allowed for improved performance characteristics of current biosensors for the detection of established analytes (glucose, pesticides etc….). Advancement has been the utilization of genetically modified enzymes to detect novel markers. An additional group of improvements is the usage of “non-traditional” transducer materials, e.g. carbon nanotubes (CNT), or different conductive polymers. Remarkable structural and electrical property advancements have enabled new options mainly in the area of electrochemical sensing technologies.

Developments are not always based on novel materials, but also on new, original approaches. One example, of such an innovation is the recently emerging field of biomolecular computing (Biocomputing). This is where enzymatic sensing systems are used to perform various computing operations that mimic processes typical of electronic computing devices. Such approaches, with their sophisticated biomolecular design, have resulted in reversible, reconfigurable, and resettable “bio-logic” sensing architectures (gates) for processing chemical information. This is especially useful in enzymatic biosensor applications where, until now, the use of biosensor or bioassay arrays have been required for a simultaneous analysis of several different species

The technology of enzymatic biosensors offers a potent combination of performance and analytical features not available in any other bioanalytical system. The listing of just a few options in this overview can encourage future development, which could yield new generations of enzymatic biosensors for a wide range of applications in clinical, environmental or industrial diagnostics.

Dr. Jan Halámek
Guest Editor

Submission

Manuscripts should be submitted online at www.mdpi.com by registering and logging in to this website. Once you are registered, click here to go to the submission form. Manuscripts can be submitted until the deadline. Papers will be published continuously (as soon as accepted) and will be listed together on the special issue website. Research articles, review articles as well as communications are invited. For planned papers, a title and short abstract (about 100 words) can be sent to the Editorial Office for announcement on this website.

Submitted manuscripts should not have been published previously, nor be under consideration for publication elsewhere (except conference proceedings papers). All manuscripts are refereed through a peer-review process. A guide for authors and other relevant information for submission of manuscripts is available on the Instructions for Authors page. Sensors is an international peer-reviewed Open Access monthly journal published by MDPI.

Please visit the Instructions for Authors page before submitting a manuscript. The Article Processing Charge (APC) for publication in this open access journal is 1800 CHF (Swiss Francs).


Published Papers (8 papers)

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Research

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Open AccessArticle Optimization of Hydrogen Peroxide Detection for a Methyl Mercaptan Biosensor
Sensors 2013, 13(4), 5028-5039; doi:10.3390/s130405028
Received: 11 March 2013 / Revised: 3 April 2013 / Accepted: 7 April 2013 / Published: 15 April 2013
Cited by 7 | PDF Full-text (282 KB) | HTML Full-text | XML Full-text
Abstract
Several kinds of modified carbon screen printed electrodes (CSPEs) for amperometric detection of hydrogen peroxide (H2O2) are presented in order to propose a methyl mercaptan (MM) biosensor. Unmodified, carbon nanotubes (CNTs), cobalt phthalocyanine (CoPC), Prussian blue (PB), and Os-wired
[...] Read more.
Several kinds of modified carbon screen printed electrodes (CSPEs) for amperometric detection of hydrogen peroxide (H2O2) are presented in order to propose a methyl mercaptan (MM) biosensor. Unmodified, carbon nanotubes (CNTs), cobalt phthalocyanine (CoPC), Prussian blue (PB), and Os-wired HRP modified CSPE sensors were fabricated and tested to detect H2O2, applying a potential of +0.6 V, +0.6 V, +0.4 V, −0.2 V and −0.1 V (versus Ag/AgCl), respectively. The limits of detection of these electrodes for H2O2 were 3.1 μM, 1.3 μM, 71 nM, 1.3 μM, 13.7 nM, respectively. The results demonstrated that the Os-wired HRP modified CSPEs gives the lowest limit of detection (LOD) for H2O2 at a working potential as low as −0.1 V. Os-wired HRP is the optimum choice for establishment of a MM biosensor and gives a detection limit of 0.5 μM. Full article
(This article belongs to the Special Issue Enzymatic Biosensors)
Open AccessArticle Carbon Based Electrodes Modified with Horseradish Peroxidase Immobilized in Conducting Polymers for Acetaminophen Analysis
Sensors 2013, 13(4), 4841-4854; doi:10.3390/s130404841
Received: 30 January 2013 / Revised: 25 March 2013 / Accepted: 8 April 2013 / Published: 11 April 2013
Cited by 8 | PDF Full-text (343 KB) | HTML Full-text | XML Full-text
Abstract
The development and optimization of new biosensors with horseradish peroxidase immobilized in carbon nanotubes-polyethyleneimine or polypyrrole nanocomposite film at the surface of two types of transducer is described. The amperometric detection of acetaminophen was carried out at −0.2 V versus Ag/AgCl using carbon
[...] Read more.
The development and optimization of new biosensors with horseradish peroxidase immobilized in carbon nanotubes-polyethyleneimine or polypyrrole nanocomposite film at the surface of two types of transducer is described. The amperometric detection of acetaminophen was carried out at −0.2 V versus Ag/AgCl using carbon based-screen printed electrodes (SPEs) and glassy carbon electrodes (GCEs) as transducers. The electroanalytical parameters of the biosensors are highly dependent on their configuration and on the dimensions of the carbon nanotubes. The best limit of detection obtained for acetaminophen was 1.36 ± 0.013 μM and the linear range 9.99–79.01 μM for the HRP-SWCNT/PEI in GCE configuration. The biosensors were successfully applied for the detection of acetaminophen in several drug formulations. Full article
(This article belongs to the Special Issue Enzymatic Biosensors)
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Open AccessArticle Internal Calibration Förster Resonance Energy Transfer Assay: A Real-Time Approach for Determining Protease Kinetics
Sensors 2013, 13(4), 4553-4570; doi:10.3390/s130404553
Received: 18 February 2013 / Revised: 11 March 2013 / Accepted: 25 March 2013 / Published: 8 April 2013
Cited by 4 | PDF Full-text (880 KB) | HTML Full-text | XML Full-text
Abstract
Förster resonance energy transfer (FRET) technology has been widely used in biological and biomedical research. This powerful tool can elucidate protein interactions in either a dynamic or steady state. We recently developed a series of FRET-based technologies to determine protein interaction dissociation constant
[...] Read more.
Förster resonance energy transfer (FRET) technology has been widely used in biological and biomedical research. This powerful tool can elucidate protein interactions in either a dynamic or steady state. We recently developed a series of FRET-based technologies to determine protein interaction dissociation constant and for use in high-throughput screening assays of SUMOylation. SUMO (small ubiquitin-like modifier) is conjugated to substrates through an enzymatic cascade. This important posttranslational protein modification is critical for multiple biological processes. Sentrin/SUMO-specific proteases (SENPs) act as endopeptidases to process the pre-SUMO or as isopeptidases to deconjugate SUMO from its substrate. Here, we describe a novel quantitative FRET-based protease assay for determining the kinetics of SENP1. Our strategy is based on the quantitative analysis and differentiation of fluorescent emission signals at the FRET acceptor emission wavelengths. Those fluorescent emission signals consist of three components: the FRET signal and the fluorescent emissions of donor (CyPet) and acceptor (YPet). Unlike our previous method in which donor and acceptor direct emissions were excluded by standard curves, the three fluorescent emissions were determined quantitatively during the SENP digestion process from onesample. New mathematical algorithms were developed to determine digested substrate concentrations directly from the FRET signal and donor/acceptor direct emissions. The kinetic parameters, kcat, KM, and catalytic efficiency (kcat/KM) of SENP1 catalytic domain for pre-SUMO1/2/3 were derived. Importantly, the general principles of this new quantitative methodology of FRET-based protease kinetic determinations can be applied to other proteases in a robust and systems biology approach. Full article
(This article belongs to the Special Issue Enzymatic Biosensors)
Open AccessArticle DNA-Based Sensor for Real-Time Measurement of the Enzymatic Activity of Human Topoisomerase I
Sensors 2013, 13(4), 4017-4028; doi:10.3390/s130404017
Received: 1 February 2013 / Revised: 16 February 2013 / Accepted: 19 March 2013 / Published: 25 March 2013
Cited by 7 | PDF Full-text (293 KB) | HTML Full-text | XML Full-text | Supplementary Files
Abstract
Sensors capable of quantitative real-time measurements may present the easiest and most accurate way to study enzyme activities. Here we present a novel DNA-based sensor for specific and quantitative real-time measurement of the enzymatic activity of the essential human enzyme, topoisomerase I. The
[...] Read more.
Sensors capable of quantitative real-time measurements may present the easiest and most accurate way to study enzyme activities. Here we present a novel DNA-based sensor for specific and quantitative real-time measurement of the enzymatic activity of the essential human enzyme, topoisomerase I. The basic design of the sensor relies on two DNA strands that hybridize to form a hairpin structure with a fluorophore-quencher pair. The quencher moiety is released from the sensor upon reaction with human topoisomerase I thus enabling real-time optical measurement of enzymatic activity. The sensor is specific for topoisomerase I even in raw cell extracts and presents a simple mean of following enzyme kinetics using standard laboratory equipment such as a qPCR machine or fluorimeter. Human topoisomerase I is a well-known target for the clinically used anti-cancer drugs of the camptothecin family. The cytotoxic effect of camptothecins correlates directly with the intracellular topoisomerase I activity. We therefore envision that the presented sensor may find use for the prediction of cellular drug response. Moreover, inhibition of topoisomerase I by camptothecin is readily detectable using the presented DNA sensor, suggesting a potential application of the sensor for first line screening for potential topoisomerase I targeting anti-cancer drugs. Full article
(This article belongs to the Special Issue Enzymatic Biosensors)
Open AccessArticle Nanobiosensors Based on Chemically Modified AFM Probes: A Useful Tool for Metsulfuron-Methyl Detection
Sensors 2013, 13(2), 1477-1489; doi:10.3390/s130201477
Received: 20 November 2012 / Revised: 6 January 2013 / Accepted: 6 January 2013 / Published: 24 January 2013
Cited by 9 | PDF Full-text (931 KB) | HTML Full-text | XML Full-text
Abstract
The use of agrochemicals has increased considerably in recent years, and consequently, there has been increased exposure of ecosystems and human populations to these highly toxic compounds. The study and development of methodologies to detect these substances with greater sensitivity has become extremely
[...] Read more.
The use of agrochemicals has increased considerably in recent years, and consequently, there has been increased exposure of ecosystems and human populations to these highly toxic compounds. The study and development of methodologies to detect these substances with greater sensitivity has become extremely relevant. This article describes, for the first time, the use of atomic force spectroscopy (AFS) in the detection of enzyme-inhibiting herbicides. A nanobiosensor based on an atomic force microscopy (AFM) tip functionalised with the acetolactate synthase (ALS) enzyme was developed and characterised. The herbicide metsulfuron-methyl, an ALS inhibitor, was successfully detected through the acquisition of force curves using this biosensor. The adhesion force values were considerably higher when the biosensor was used. An increase of ~250% was achieved relative to the adhesion force using an unfunctionalised AFM tip. This considerable increase was the result of a specific interaction between the enzyme and the herbicide, which was primarily responsible for the efficiency of the nanobiosensor. These results indicate that this methodology is promising for the detection of herbicides, pesticides, and other environmental contaminants. Full article
(This article belongs to the Special Issue Enzymatic Biosensors)
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Open AccessArticle Indirect Determination of Mercury Ion by Inhibition of a Glucose Biosensor Based on ZnO Nanorods
Sensors 2012, 12(11), 15063-15077; doi:10.3390/s121115063
Received: 7 September 2012 / Revised: 17 October 2012 / Accepted: 2 November 2012 / Published: 6 November 2012
Cited by 17 | PDF Full-text (707 KB) | HTML Full-text | XML Full-text
Abstract
A potentiometric glucose biosensor based on immobilization of glucose oxidase (GOD) on ZnO nanorods (ZnO-NRs) has been developed for the indirect determination of environmental mercury ions. The ZnO-NRs were grown on a gold coated glass substrate by using the low temperature aqueous chemical
[...] Read more.
A potentiometric glucose biosensor based on immobilization of glucose oxidase (GOD) on ZnO nanorods (ZnO-NRs) has been developed for the indirect determination of environmental mercury ions. The ZnO-NRs were grown on a gold coated glass substrate by using the low temperature aqueous chemical growth (ACG) approach. Glucose oxidase in conjunction with a chitosan membrane and a glutaraldehyde (GA) were immobilized on the surface of the ZnO-NRs using a simple physical adsorption method and then used as a potentiometric working electrode. The potential response of the biosensor between the working electrode and an Ag/AgCl reference electrode was measured in a 1mM phosphate buffer solution (PBS). The detection limit of the mercury ion sensor was found to be 0.5 nM. The experimental results provide two linear ranges of the inhibition from 0.5 × 10−6 mM to 0.5 × 10−4 mM, and from 0.5 × 10−4 mM to 20 mM of mercury ion for fixed 1 mM of glucose concentration in the solution. The linear range of the inhibition from 10−3 mM to 6 mM of mercury ion was also acquired for a fixed 10 mM of glucose concentration. The working electrode can be reactivated by more than 70% after inhibition by simply dipping the used electrode in a 10 mM PBS solution for 7 min. The electrodes retained their original enzyme activity by about 90% for more than three weeks. The response to mercury ions was highly sensitive, selective, stable, reproducible, and interference resistant, and exhibits a fast response time. The developed glucose biosensor has a great potential for detection of mercury with several advantages such as being inexpensive, requiring minimum hardware and being suitable for unskilled users. Full article
(This article belongs to the Special Issue Enzymatic Biosensors)
Open AccessArticle Catalytic and Inhibitory Kinetic Behavior of Horseradish Peroxidase on the Electrode Surface
Sensors 2012, 12(11), 14556-14569; doi:10.3390/s121114556
Received: 2 August 2012 / Revised: 29 August 2012 / Accepted: 17 September 2012 / Published: 29 October 2012
Cited by 1 | PDF Full-text (480 KB) | HTML Full-text | XML Full-text
Abstract
Enzymatic biosensors are often used to detect trace levels of some specific substance. An alternative methodology is applied for enzymatic assays, in which the electrocatalytic kinetic behavior of enzymes is monitored by measuring the faradaic current for a variety of substrate and inhibitor
[...] Read more.
Enzymatic biosensors are often used to detect trace levels of some specific substance. An alternative methodology is applied for enzymatic assays, in which the electrocatalytic kinetic behavior of enzymes is monitored by measuring the faradaic current for a variety of substrate and inhibitor concentrations. Here we examine a steady-state and pre-steady-state reduction of H2O2 on the horseradish peroxidase electrode. The results indicate the substrate-concentration dependence of the steady-state current strictly obeys Michaelis-Menten kinetics rules; in other cases there is ambiguity, whereby he inhibitor-concentration dependence of the steady-state current has a discontinuity under moderate concentration conditions. For pre-steady-state phases, both catalysis and inhibition show an abrupt change of the output current. These anomalous phenomena are universal and there might be an underlying biochemical or electrochemical rationale. Full article
(This article belongs to the Special Issue Enzymatic Biosensors)

Review

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Open AccessReview Immobilization Techniques in the Fabrication of Nanomaterial-Based Electrochemical Biosensors: A Review
Sensors 2013, 13(4), 4811-4840; doi:10.3390/s130404811
Received: 1 March 2013 / Revised: 2 April 2013 / Accepted: 9 April 2013 / Published: 11 April 2013
Cited by 71 | PDF Full-text (688 KB) | HTML Full-text | XML Full-text
Abstract
The evolution of 1st to 3rd generation electrochemical biosensors reflects a simplification and enhancement of the transduction pathway. However, in recent years, modification of the transducer with nanomaterials has become increasingly studied and imparts many advantages. The sensitivity and overall performance of enzymatic
[...] Read more.
The evolution of 1st to 3rd generation electrochemical biosensors reflects a simplification and enhancement of the transduction pathway. However, in recent years, modification of the transducer with nanomaterials has become increasingly studied and imparts many advantages. The sensitivity and overall performance of enzymatic biosensors has improved tremendously as a result of incorporating nanomaterials in their fabrication. Given the unique and favorable qualities of gold nanoparticles, graphene and carbon nanotubes as applied to electrochemical biosensors, a consolidated survey of the different methods of nanomaterial immobilization on transducer surfaces and enzyme immobilization on these species is beneficial and timely. This review encompasses modification of enzymatic biosensors with gold nanoparticles, carbon nanotubes, and graphene. Full article
(This article belongs to the Special Issue Enzymatic Biosensors)

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