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24 pages, 4215 KiB

Open AccessArticle

The Scavenging Effect of Myoglobin from Meat Extracts toward Peroxynitrite Studied with a Flow Injection System Based on Electrochemical Reduction over a Screen-Printed Carbon Electrode Modified with Cobalt Phthalocyanine: Quantification and Kinetics

by Ioana Silvia Hosu, Diana Constantinescu-Aruxandei, Florin Oancea and Mihaela Doni

Biosensors 2021, 11(7), 220; https://doi.org/10.3390/bios11070220 - 02 Jul 2021

Viewed by 2971

Abstract

The scavenging activity of myoglobin toward peroxynitrite (PON) was studied in meat extracts, using a new developed electrochemical method (based on cobalt phthalocyanine-modified screen-printed carbon electrode, SPCE/CoPc) and calculating kinetic parameters of PON decay (such as half-time and apparent rate constants). As reactive [...] Read more.

The scavenging activity of myoglobin toward peroxynitrite (PON) was studied in meat extracts, using a new developed electrochemical method (based on cobalt phthalocyanine-modified screen-printed carbon electrode, SPCE/CoPc) and calculating kinetic parameters of PON decay (such as half-time and apparent rate constants). As reactive oxygen/nitrogen species (ROS/RNS) affect the food quality, the consumers can be negatively influenced. The discoloration, rancidity, and flavor of meat are altered in the presence of these species, such as PON. Our new highly thermically stable, cost-effective, rapid, and simple electrocatalytical method was combined with a flow injection analysis system to achieve high sensitivity (10.843 nA µM⁻¹) at a nanomolar level LoD (400 nM), within a linear range of 3–180 µM. The proposed biosensor was fully characterized using SEM, FTIR, Raman spectroscopy, Cyclic Voltammetry (CV), Differential Pulse Voltammetry (DPV), and Linear Sweep Voltammetry (LSV). These achievements were obtained due to the CoPc-mediated reduction of PON at very low potentials (around 0.1 V vs. Ag/AgCl pseudoreference). We also proposed a redox mechanism involving two electrons in the reduction of peroxynitrite to nitrite and studied some important interfering species (nitrite, nitrate, hydrogen peroxide, dopamine, ascorbic acid), which showed that our method is highly selective. These features make our work relevant, as it could be further applied to study the kinetics of important oxidative processes in vivo or in vitro, as PON is usually present in the nanomolar or micromolar range in physiological conditions, and our method is sensitive enough to be applied. Full article

(This article belongs to the Special Issue Selected Papers from the 1st International Electronic Conference on Biosensors (IECB 2020))

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18 pages, 12349 KiB

Open AccessArticle

“Green” Prussian Blue Analogues as Peroxidase Mimetics for Amperometric Sensing and Biosensing

by Galina Z. Gayda, Olha M. Demkiv, Yanna Gurianov, Roman Ya. Serkiz, Halyna M. Klepach, Mykhailo V. Gonchar and Marina Nisnevitch

Biosensors 2021, 11(6), 193; https://doi.org/10.3390/bios11060193 - 10 Jun 2021

Cited by 9 | Viewed by 3327

Abstract

Prussian blue analogs (PBAs) are well-known artificial enzymes with peroxidase (PO)-like activity. PBAs have a high potential for applications in scientific investigations, industry, ecology and medicine. Being stable and both catalytically and electrochemically active, PBAs are promising in the construction of biosensors and [...] Read more.

Prussian blue analogs (PBAs) are well-known artificial enzymes with peroxidase (PO)-like activity. PBAs have a high potential for applications in scientific investigations, industry, ecology and medicine. Being stable and both catalytically and electrochemically active, PBAs are promising in the construction of biosensors and biofuel cells. The “green” synthesis of PO-like PBAs using oxido-reductase flavocytochrome b₂ is described in this study. When immobilized on graphite electrodes (GEs), the obtained green-synthesized PBAs or hexacyanoferrates (gHCFs) of transition and noble metals produced amperometric signals in response to H₂O₂. HCFs of copper, iron, palladium and other metals were synthesized and characterized by structure, size, catalytic properties and electro-mediator activities. The gCuHCF, as the most effective PO mimetic with a flower-like micro/nano superstructure, was used as an H₂O₂-sensitive platform for the development of a glucose oxidase (GO)-based biosensor. The GO/gCuHCF/GE biosensor exhibited high sensitivity (710 A M⁻¹m⁻²), a broad linear range and good selectivity when tested on real samples of fruit juices. We propose that the gCuHCF and other gHCFs synthesized via enzymes may be used as artificial POs in amperometric oxidase-based (bio)sensors. Full article

(This article belongs to the Special Issue Selected Papers from the 1st International Electronic Conference on Biosensors (IECB 2020))

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14 pages, 2334 KiB

Open AccessArticle

Surface Plasmon Resonance Assay for Label-Free and Selective Detection of HIV-1 p24 Protein

by Lucia Sarcina, Giuseppe Felice Mangiatordi, Fabrizio Torricelli, Paolo Bollella, Zahra Gounani, Ronald Österbacka, Eleonora Macchia and Luisa Torsi

Biosensors 2021, 11(6), 180; https://doi.org/10.3390/bios11060180 - 03 Jun 2021

Cited by 15 | Viewed by 4003

Abstract

The early detection of the human immunodeficiency virus (HIV) is of paramount importance to achieve efficient therapeutic treatment and limit the disease spreading. In this perspective, the assessment of biosensing assay for the HIV-1 p24 capsid protein plays a pivotal role in the [...] Read more.

The early detection of the human immunodeficiency virus (HIV) is of paramount importance to achieve efficient therapeutic treatment and limit the disease spreading. In this perspective, the assessment of biosensing assay for the HIV-1 p24 capsid protein plays a pivotal role in the timely and selective detection of HIV infections. In this study, multi-parameter-SPR has been used to develop a reliable and label-free detection method for HIV-1 p24 protein. Remarkably, both physical and chemical immobilization of mouse monoclonal antibodies against HIV-1 p24 on the SPR gold detecting surface have been characterized for the first time. The two immobilization techniques returned a capturing antibody surface coverage as high as (7.5 ± 0.3) × 10¹¹ molecule/cm² and (2.4 ± 0.6) × 10¹¹ molecule/cm², respectively. However, the covalent binding of the capturing antibodies through a mixed self-assembled monolayer (SAM) of alkanethiols led to a doubling of the p24 binding signal. Moreover, from the modeling of the dose-response curve, an equilibrium dissociation constant K_D of 5.30 × 10⁻⁹ M was computed for the assay performed on the SAM modified surface compared to a much larger K_D of 7.46 × 10⁻⁵ M extracted for the physisorbed antibodies. The chemically modified system was also characterized in terms of sensitivity and selectivity, reaching a limit of detection of (4.1 ± 0.5) nM and an unprecedented selectivity ratio of 0.02. Full article

(This article belongs to the Special Issue Selected Papers from the 1st International Electronic Conference on Biosensors (IECB 2020))

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14 pages, 2444 KiB

Open AccessEditor’s ChoiceArticle

Light-Addressable Actuator-Sensor Platform for Monitoring and Manipulation of pH Gradients in Microfluidics: A Case Study with the Enzyme Penicillinase

by Rene Welden, Melanie Jablonski, Christina Wege, Michael Keusgen, Patrick Hermann Wagner, Torsten Wagner and Michael J. Schöning

Biosensors 2021, 11(6), 171; https://doi.org/10.3390/bios11060171 - 27 May 2021

Cited by 19 | Viewed by 3130

Abstract

The feasibility of light-addressed detection and manipulation of pH gradients inside an electrochemical microfluidic cell was studied. Local pH changes, induced by a light-addressable electrode (LAE), were detected using a light-addressable potentiometric sensor (LAPS) with different measurement modes representing an actuator-sensor system. Biosensor [...] Read more.

The feasibility of light-addressed detection and manipulation of pH gradients inside an electrochemical microfluidic cell was studied. Local pH changes, induced by a light-addressable electrode (LAE), were detected using a light-addressable potentiometric sensor (LAPS) with different measurement modes representing an actuator-sensor system. Biosensor functionality was examined depending on locally induced pH gradients with the help of the model enzyme penicillinase, which had been immobilized in the microfluidic channel. The surface morphology of the LAE and enzyme-functionalized LAPS was studied by scanning electron microscopy. Furthermore, the penicillin sensitivity of the LAPS inside the microfluidic channel was determined with regard to the analyte’s pH influence on the enzymatic reaction rate. In a final experiment, the LAE-controlled pH inhibition of the enzyme activity was monitored by the LAPS. Full article

(This article belongs to the Special Issue Selected Papers from the 1st International Electronic Conference on Biosensors (IECB 2020))

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13 pages, 4788 KiB

Open AccessArticle

Comparison of Leading Biosensor Technologies to Detect Changes in Human Endothelial Barrier Properties in Response to Pro-Inflammatory TNFα and IL1β in Real-Time

by James J. W. Hucklesby, Akshata Anchan, Simon J. O'Carroll, Charles P. Unsworth, E. Scott Graham and Catherine E. Angel

Biosensors 2021, 11(5), 159; https://doi.org/10.3390/bios11050159 - 18 May 2021

Cited by 7 | Viewed by 2599

Abstract

Electric Cell-Substrate Impedance Sensing (ECIS), xCELLigence and cellZscope are commercially available instruments that measure the impedance of cellular monolayers. Despite widespread use of these systems individually, direct comparisons between these platforms have not been published. To compare these instruments, the responses of human [...] Read more.

Electric Cell-Substrate Impedance Sensing (ECIS), xCELLigence and cellZscope are commercially available instruments that measure the impedance of cellular monolayers. Despite widespread use of these systems individually, direct comparisons between these platforms have not been published. To compare these instruments, the responses of human brain endothelial monolayers to TNFα and IL1β were measured on all three platforms simultaneously. All instruments detected transient changes in impedance in response to the cytokines, although the response magnitude varied, with ECIS being the most sensitive. ECIS and cellZscope were also able to attribute responses to particular endothelial barrier components by modelling the multifrequency impedance data acquired by these instruments; in contrast the limited frequency xCELLigence data cannot be modelled. Consistent with its superior impedance sensing, ECIS exhibited a greater capacity than cellZscope to distinguish between subtle changes in modelled endothelial monolayer properties. The reduced resolving ability of the cellZscope platform may be due to its electrode configuration, which is necessary to allow access to the basolateral compartment, an important advantage of this instrument. Collectively, this work demonstrates that instruments must be carefully selected to ensure they are appropriate for the experimental questions being asked when assessing endothelial barrier properties. Full article

(This article belongs to the Special Issue Selected Papers from the 1st International Electronic Conference on Biosensors (IECB 2020))

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16 pages, 1826 KiB

Open AccessArticle

Fast and Sensitive Determination of the Fungicide Carbendazim in Fruit Juices with an Immunosensor Based on White Light Reflectance Spectroscopy

by Georgios Koukouvinos, Chrysoula-Evangelia Karachaliou, Ioannis Raptis, Panagiota Petrou, Evangelia Livaniou and Sotirios Kakabakos

Biosensors 2021, 11(5), 153; https://doi.org/10.3390/bios11050153 - 13 May 2021

Cited by 6 | Viewed by 2506

Abstract

Carbendazim is a systemic benzimidazole-type fungicide with broad-spectrum activity against fungi that undermine food products safety and quality. Despite its effectiveness, carbendazim constitutes a major environmental pollutant, being hazardous to both humans and animals. Therefore, fast and reliable determination of carbendazim levels in [...] Read more.

Carbendazim is a systemic benzimidazole-type fungicide with broad-spectrum activity against fungi that undermine food products safety and quality. Despite its effectiveness, carbendazim constitutes a major environmental pollutant, being hazardous to both humans and animals. Therefore, fast and reliable determination of carbendazim levels in water, soil, and food samples is of high importance for both food industry and public health. Herein, an optical biosensor based on white light reflectance spectroscopy (WLRS) for fast and sensitive determination of carbendazim in fruit juices is presented. The transducer is a Si/SiO₂ chip functionalized with a benzimidazole conjugate, and determination is based on a competitive immunoassay format. Thus, for the assay, a mixture of an in-house developed rabbit polyclonal anti-carbendazim antibody with the standards or samples is pumped over the chip, followed by biotinylated secondary antibody and streptavidin. The WLRS platform allows for real-time monitoring of biomolecular interactions carried out onto the Si/SiO₂ chip by transforming the shift in the reflected interference spectrum caused by the immunoreaction to effective biomolecular adlayer thickness. The sensor is able to detect 20 ng/mL of carbendazim in fruit juices with high accuracy and precision (intra- and inter-assay CVs ≤ 6.9% and ≤9.4%, respectively) in less than 30 min, applying a simple sample treatment that alleviates any “matrix-effect” on the assay results and a 60 min preincubation step for improving assay sensitivity. Excellent analytical characteristics and short analysis time along with its small size render the proposed WLRS immunosensor ideal for future on-the-spot determination of carbendazim in food and environmental samples. Full article

(This article belongs to the Special Issue Selected Papers from the 1st International Electronic Conference on Biosensors (IECB 2020))

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17 pages, 2809 KiB

Open AccessArticle

Detection of Sub-Nanomolar Concentration of Trypsin by Thickness-Shear Mode Acoustic Biosensor and Spectrophotometry

by Ivan Piovarci, Sopio Melikishvili, Marek Tatarko, Tibor Hianik and Michael Thompson

Biosensors 2021, 11(4), 117; https://doi.org/10.3390/bios11040117 - 11 Apr 2021

Cited by 14 | Viewed by 2907

Abstract

The determination of protease activity is very important for disease diagnosis, drug development, and quality and safety assurance for dairy products. Therefore, the development of low-cost and sensitive methods for assessing protease activity is crucial. We report two approaches for monitoring protease activity: [...] Read more.

The determination of protease activity is very important for disease diagnosis, drug development, and quality and safety assurance for dairy products. Therefore, the development of low-cost and sensitive methods for assessing protease activity is crucial. We report two approaches for monitoring protease activity: in a volume and at surface, via colorimetric and acoustic wave-based biosensors operated in the thickness-shear mode (TSM), respectively. The TSM sensor was based on a β-casein substrate immobilized on a piezoelectric quartz crystal transducer. After an enzymatic reaction with trypsin, it cleaved the surface-bound β-casein, which increased the resonant frequency of the crystal. The limit of detection (LOD) was 0.48 ± 0.08 nM. A label-free colorimetric assay for trypsin detection has also been performed using β-casein and 6-mercaptohexanol (MCH) functionalized gold nanoparticles (AuNPs/MCH-β-casein). Due to the trypsin cleavage of β-casein, the gold nanoparticles lost shelter, and MCH increased the attractive force between the modified AuNPs. Consequently, AuNPs aggregated, and the red shift of the absorption spectra was observed. Spectrophotometric assay enabled an LOD of 0.42 ± 0.03 nM. The Michaelis–Menten constant, K_M, for reverse enzyme reaction has also been estimated by both methods. This value for the colorimetric assay (0.56 ± 0.10 nM) is lower in comparison with those for the TSM sensor (0.92 ± 0.44 nM). This is likely due to the better access of the trypsin to the β-casein substrate at the surface of AuNPs in comparison with those at the TSM transducer. Full article

(This article belongs to the Special Issue Selected Papers from the 1st International Electronic Conference on Biosensors (IECB 2020))

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17 pages, 3546 KiB

Open AccessArticle

Contactless Temperature Sensing at the Microscale Based on Titanium Dioxide Raman Thermometry

by Veronica Zani, Danilo Pedron, Roberto Pilot and Raffaella Signorini

Biosensors 2021, 11(4), 102; https://doi.org/10.3390/bios11040102 - 02 Apr 2021

Cited by 7 | Viewed by 2943

Abstract

The determination of local temperature at the nanoscale is a key point to govern physical, chemical and biological processes, strongly influenced by temperature. Since a wide range of applications, from nanomedicine to nano- or micro-electronics, requires a precise determination of the local temperature, [...] Read more.

The determination of local temperature at the nanoscale is a key point to govern physical, chemical and biological processes, strongly influenced by temperature. Since a wide range of applications, from nanomedicine to nano- or micro-electronics, requires a precise determination of the local temperature, significant efforts have to be devoted to nanothermometry. The identification of efficient materials and the implementation of detection techniques are still a hot topic in nanothermometry. Many strategies have been already investigated and applied to real cases, but there is an urgent need to develop new protocols allowing for accurate and sensitive temperature determination. The focus of this work is the investigation of efficient optical thermometers, with potential applications in the biological field. Among the different optical techniques, Raman spectroscopy is currently emerging as a very interesting tool. Its main advantages rely on the possibility of carrying out non-destructive and non-contact measurements with high spatial resolution, reaching even the nanoscale. Temperature variations can be determined by following the changes in intensity, frequency position and width of one or more bands. Concerning the materials, Titanium dioxide has been chosen as Raman active material because of its intense cross-section and its biocompatibility, as already demonstrated in literature. Raman measurements have been performed on commercial anatase powder, with a crystallite dimension of hundreds of nm, using 488.0, 514.5, 568.2 and 647.1 nm excitation lines of the CW Ar⁺/Kr⁺ ion laser. The laser beam was focalized through a microscope on the sample, kept at defined temperature using a temperature controller, and the temperature was varied in the range of 283–323 K. The Stokes and anti-Stokes scattered light was analyzed through a triple monochromator and detected by a liquid nitrogen-cooled CCD camera. Raw data have been analyzed with Matlab, and Raman spectrum parameters—such as area, intensity, frequency position and width of the peak—have been calculated using a Lorentz fitting curve. Results obtained, calculating the anti-Stokes/Stokes area ratio, demonstrate that the Raman modes of anatase, in particular the E_g one at 143 cm⁻¹, are excellent candidates for the local temperature detection in the visible range. Full article

(This article belongs to the Special Issue Selected Papers from the 1st International Electronic Conference on Biosensors (IECB 2020))

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22 pages, 3215 KiB

Open AccessArticle

Market Perspectives and Future Fields of Application of Odor Detection Biosensors within the Biological Transformation—A Systematic Analysis

by Johannes Full, Yannick Baumgarten, Lukas Delbrück, Alexander Sauer and Robert Miehe

Biosensors 2021, 11(3), 93; https://doi.org/10.3390/bios11030093 - 23 Mar 2021

Cited by 12 | Viewed by 3579

Abstract

The technological advantages that biosensors have over conventional technical sensors for odor detection and the role they play in the biological transformation have not yet been comprehensively analyzed. However, this is necessary for assessing their suitability for specific fields of application as well [...] Read more.

The technological advantages that biosensors have over conventional technical sensors for odor detection and the role they play in the biological transformation have not yet been comprehensively analyzed. However, this is necessary for assessing their suitability for specific fields of application as well as their improvement and development goals. An overview of biological basics of olfactory systems is given and different odor sensor technologies are described and classified in this paper. Specific market potentials of biosensors for odor detection are identified by applying a tailored methodology that enables the derivation and systematic comparison of both the performance profiles of biosensors as well as the requirement profiles for various application fields. Therefore, the fulfillment of defined requirements is evaluated for biosensors by means of 16 selected technical criteria in order to determine a specific performance profile. Further, a selection of application fields, namely healthcare, food industry, agriculture, cosmetics, safety applications, environmental monitoring for odor detection sensors is derived to compare the importance of the criteria for each of the fields, leading to market-specific requirement profiles. The analysis reveals that the requirement criteria considered to be the most important ones across all application fields are high specificity, high selectivity, high repeat accuracy, high resolution, high accuracy, and high sensitivity. All these criteria, except for the repeat accuracy, can potentially be better met by biosensors than by technical sensors, according to the results obtained. Therefore, biosensor technology in general has a high application potential for all the areas of application under consideration. Health and safety applications especially are considered to have high potential for biosensors due to their correspondence between requirement and performance profiles. Special attention is paid to new areas of application that require multi-sensing capability. Application scenarios for multi-sensing biosensors are therefore derived. Moreover, the role of biosensors within the biological transformation is discussed. Full article

(This article belongs to the Special Issue Selected Papers from the 1st International Electronic Conference on Biosensors (IECB 2020))

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12 pages, 1998 KiB

Open AccessCommunication

Microfluidic Impedance Biosensor Chips Using Sensing Layers Based on DNA-Based Self-Assembled Monolayers for Label-Free Detection of Proteins

by Khaled Alsabbagh, Tim Hornung, Achim Voigt, Sahba Sadir, Taleieh Rajabi and Kerstin Länge

Biosensors 2021, 11(3), 80; https://doi.org/10.3390/bios11030080 - 13 Mar 2021

Cited by 14 | Viewed by 3119

Abstract

A microfluidic chip for electrochemical impedance spectroscopy (EIS) is presented as bio-sensor for label-free detection of proteins by using the example of cardiac troponin I. Troponin I is one of the most specific diagnostic serum biomarkers for myocardial infarction. The microfluidic impedance biosensor [...] Read more.

A microfluidic chip for electrochemical impedance spectroscopy (EIS) is presented as bio-sensor for label-free detection of proteins by using the example of cardiac troponin I. Troponin I is one of the most specific diagnostic serum biomarkers for myocardial infarction. The microfluidic impedance biosensor chip presented here consists of a microscope glass slide serving as base plate, sputtered electrodes, and a polydimethylsiloxane (PDMS) microchannel. Electrode functionalization protocols were developed considering a possible charge transfer through the sensing layer, in addition to analyte-specific binding by corresponding antibodies and reduction of nonspecific protein adsorption to prevent false-positive signals. Reagents tested for self-assembled monolayers (SAMs) on gold electrodes included thiolated hydrocarbons and thiolated oligonucleotides, where SAMs based on the latter showed a better performance. The corresponding antibody was covalently coupled on the SAM using carbodiimide chemistry. Sampling and measurement took only a few minutes. Application of a human serum albumin (HSA) sample, 1000 ng/mL, led to negligible impedance changes, while application of a troponin I sample, 1 ng/mL, led to a significant shift in the Nyquist plot. The results are promising regarding specific detection of clinically relevant concentrations of biomarkers, such as cardiac markers, with the newly developed microfluidic impedance biosensor chip. Full article

(This article belongs to the Special Issue Selected Papers from the 1st International Electronic Conference on Biosensors (IECB 2020))

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12 pages, 4267 KiB

Open AccessArticle

SPR-Optical Fiber-Molecularly Imprinted Polymer Sensor for the Detection of Furfural in Wine

by Maria Pesavento, Luigi Zeni, Letizia De Maria, Giancarla Alberti and Nunzio Cennamo

Biosensors 2021, 11(3), 72; https://doi.org/10.3390/bios11030072 - 05 Mar 2021

Cited by 38 | Viewed by 3717

Abstract

A surface plasmon resonance (SPR) platform, based on a D-shaped plastic optical fiber (POF), combined with a biomimetic receptor, i.e., a molecularly imprinted polymer (MIP), is proposed to detect furfural (2-furaldheide, 2-FAL) in fermented beverages like wine. MIPs have been demonstrated to be [...] Read more.

A surface plasmon resonance (SPR) platform, based on a D-shaped plastic optical fiber (POF), combined with a biomimetic receptor, i.e., a molecularly imprinted polymer (MIP), is proposed to detect furfural (2-furaldheide, 2-FAL) in fermented beverages like wine. MIPs have been demonstrated to be a very convenient biomimetic receptor in the proposed sensing device, being easy and rapid to develop, suitable for on-site determinations at low concentrations, and cheap. Moreover, the MIP film thickness can be changed to modulate the sensing parameters. The possibility of performing single drop measurements is a further favorable aspect for practical applications. For example, the use of an SPR-MIP sensor for the analysis of 2-FAL in a real life matrix such as wine is proposed, obtaining a low detection limit of 0.004 mg L⁻¹. The determination of 2-FAL in fermented beverages is becoming a crucial task, mainly for the effects of the furanic compounds on the flavor of food and their toxic and carcinogenic effect on human beings. Full article

(This article belongs to the Special Issue Selected Papers from the 1st International Electronic Conference on Biosensors (IECB 2020))

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16 pages, 3293 KiB

Open AccessArticle

Electronic Eye Based on RGB Analysis for the Identification of Tequilas

by Anais Gómez, Diana Bueno and Juan Manuel Gutiérrez

Biosensors 2021, 11(3), 68; https://doi.org/10.3390/bios11030068 - 02 Mar 2021

Cited by 12 | Viewed by 3372

Abstract

The present work reports the development of a biologically inspired analytical system known as Electronic Eye (EE), capable of qualitatively discriminating different tequila categories. The reported system is a low-cost and portable instrumentation based on a Raspberry Pi single-board computer and an 8 [...] Read more.

The present work reports the development of a biologically inspired analytical system known as Electronic Eye (EE), capable of qualitatively discriminating different tequila categories. The reported system is a low-cost and portable instrumentation based on a Raspberry Pi single-board computer and an 8 Megapixel CMOS image sensor, which allow the collection of images of Silver, Aged, and Extra-aged tequila samples. Image processing is performed mimicking the trichromatic theory of color vision using an analysis of Red, Green, and Blue components (RGB) for each image’s pixel. Consequently, RGB absorbances of images were evaluated and preprocessed, employing Principal Component Analysis (PCA) to visualize data clustering. The resulting PCA scores were modeled with a Linear Discriminant Analysis (LDA) that accomplished the qualitative classification of tequilas. A Leave-One-Out Cross-Validation (LOOCV) procedure was performed to evaluate classifiers’ performance. The proposed system allowed the identification of real tequila samples achieving an overall classification rate of 90.02%, average sensitivity, and specificity of 0.90 and 0.96, respectively, while Cohen’s kappa coefficient was 0.87. In this case, the EE has demonstrated a favorable capability to correctly discriminated and classified the different tequila samples according to their categories. Full article

(This article belongs to the Special Issue Selected Papers from the 1st International Electronic Conference on Biosensors (IECB 2020))

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14 pages, 2853 KiB

Open AccessArticle

Detection of Chymotrypsin by Optical and Acoustic Methods

by Ivan Piovarci, Tibor Hianik and Ilia N. Ivanov

Biosensors 2021, 11(3), 63; https://doi.org/10.3390/bios11030063 - 26 Feb 2021

Cited by 10 | Viewed by 2992

Abstract

Chymotrypsin is an important proteolytic enzyme in the human digestive system that cleaves milk proteins through the hydrolysis reaction, making it an interesting subject to study the activity of milk proteases. In this work, we compared detection of chymotrypsin by spectrophotometric dynamic light [...] Read more.

Chymotrypsin is an important proteolytic enzyme in the human digestive system that cleaves milk proteins through the hydrolysis reaction, making it an interesting subject to study the activity of milk proteases. In this work, we compared detection of chymotrypsin by spectrophotometric dynamic light scattering (DLS) and quartz crystal microbalance (QCM) methods and determined the limit of chymotrypsin detection (LOD), 0.15 ± 0.01 nM for spectrophotometric, 0.67 ± 0.05 nM for DLS and 1.40 ± 0.30 nM for QCM methods, respectively. The sensors are relatively cheap and are able to detect chymotrypsin in 3035 min. While the optical detection methods are simple to implement, the QCM method is more robust for sample preparation, and allows detection of chymotrypsin in non-transparent samples. We give an overview on methods and instruments for detection of chymotrypsin and other milk proteases. Full article

(This article belongs to the Special Issue Selected Papers from the 1st International Electronic Conference on Biosensors (IECB 2020))

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16 pages, 2511 KiB

Open AccessArticle

Non-Invasive Determination of Glucose Concentration Using a Near-Field Sensor

by Aleksandr Gorst, Kseniya Zavyalova and Aleksandr Mironchev

Biosensors 2021, 11(3), 62; https://doi.org/10.3390/bios11030062 - 26 Feb 2021

Cited by 8 | Viewed by 2513

Abstract

The article presents a model of a near-field sensor for non-invasive glucose monitoring. The sensor has a specific design and forms a rather extended near-field. Due to this, the high penetration of electromagnetic waves into highly absorbing media (biologic media) is achieved. It [...] Read more.

The article presents a model of a near-field sensor for non-invasive glucose monitoring. The sensor has a specific design and forms a rather extended near-field. Due to this, the high penetration of electromagnetic waves into highly absorbing media (biologic media) is achieved. It represents a combined slot antenna based on a flexible RO3003 substrate. Moreover, it is small and rather flat (25 mm in diameter, 0.76 mm thick). These circumstances are the distinguishing features of this sensor in comparison with microwave sensors of other designs. The article presents the results of numerical modeling and experimental verification of a near-field sensor. Furthermore, a phantom of human biological media (human hand) was created for experimentation. In the case of numerical modeling, the sensor is located close to the hand model. In a full-scale experiment, it is located close to the phantom of the human hand for the maximum interaction of the near-field with biological materials. As a result of a series of measurements for this sensor, the reflection coefficient is measured, and the dependences of the reflected signal on the frequency are plotted. According to the results of the experiments carried out, there is a clear difference in glucose concentrations. At the same time, the accuracy of determining the difference in glucose concentrations is high. The values of the amplitude of the reflected signal with a change in concentration differ by 0.5–0.8 dB. This sensor can be used for developing a non-invasive blood glucose measurement procedure. Full article

(This article belongs to the Special Issue Selected Papers from the 1st International Electronic Conference on Biosensors (IECB 2020))

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20 pages, 3433 KiB

Open AccessArticle

Electrical Characterization of Cellulose-Based Membranes towards Pathogen Detection in Water

by Grégoire Le Brun, Margo Hauwaert, Audrey Leprince, Karine Glinel, Jacques Mahillon and Jean-Pierre Raskin

Biosensors 2021, 11(2), 57; https://doi.org/10.3390/bios11020057 - 21 Feb 2021

Cited by 6 | Viewed by 3310

Abstract

Paper substrates are promising for development of cost-effective and efficient point-of-care biosensors, essential for public healthcare and environmental diagnostics in emergency situations. Most paper-based biosensors rely on the natural capillarity of paper to perform qualitative or semi-quantitative colorimetric detections. To achieve quantification and [...] Read more.

Paper substrates are promising for development of cost-effective and efficient point-of-care biosensors, essential for public healthcare and environmental diagnostics in emergency situations. Most paper-based biosensors rely on the natural capillarity of paper to perform qualitative or semi-quantitative colorimetric detections. To achieve quantification and better sensitivity, technologies combining paper-based substrates and electrical detection are being developed. In this work, we demonstrate the potential of electrical measurements by means of a simple, parallel-plate electrode setup towards the detection of whole-cell bacteria captured in nitrocellulose (NC) membranes. Unlike current electrical sensors, which are mostly integrated, this plug and play system has reusable electrodes and enables simple and fast bacterial detection through impedance measurements. The characterized NC membrane was subjected to (i) a biofunctionalization, (ii) different saline solutions modelling real water samples, and (iii) bacterial suspensions of different concentrations. Bacterial detection was achieved in low conductivity buffers through both resistive and capacitive changes in the sensed medium. To capture Bacillus thuringiensis, the model microorganism used in this work, the endolysin cell-wall binding domain (CBD) of Deep-Blue, a bacteriophage targeting this bacterium, was integrated into the membranes as a recognition bio-interface. This experimental proof-of-concept illustrates the electrical detection of 10⁷ colony-forming units (CFU) mL⁻¹ bacteria in low-salinity buffers within 5 min, using a very simple setup. This offers perspectives for affordable pathogen sensors that can easily be reconfigured for different bacteria. Water quality testing is a particularly interesting application since it requires frequent testing, especially in emergency situations. Full article

(This article belongs to the Special Issue Selected Papers from the 1st International Electronic Conference on Biosensors (IECB 2020))

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15 pages, 1330 KiB

Open AccessArticle

Optimisation of an Electrochemical DNA Sensor for Measuring KRAS G12D and G13D Point Mutations in Different Tumour Types

by Bukola Attoye, Matthew J. Baker, Fiona Thomson, Chantevy Pou and Damion K. Corrigan

Biosensors 2021, 11(2), 42; https://doi.org/10.3390/bios11020042 - 05 Feb 2021

Cited by 12 | Viewed by 3571

Abstract

Circulating tumour DNA (ctDNA) is widely used in liquid biopsies due to having a presence in the blood that is typically in proportion to the stage of the cancer and because it may present a quick and practical method of capturing tumour heterogeneity. [...] Read more.

Circulating tumour DNA (ctDNA) is widely used in liquid biopsies due to having a presence in the blood that is typically in proportion to the stage of the cancer and because it may present a quick and practical method of capturing tumour heterogeneity. This paper outlines a simple electrochemical technique adapted towards point-of-care cancer detection and treatment monitoring from biofluids using a label-free detection strategy. The mutations used for analysis were the KRAS G12D and G13D mutations, which are both important in the initiation, progression and drug resistance of many human cancers, leading to a high mortality rate. A low-cost DNA sensor was developed to specifically investigate these common circulating tumour markers. Initially, we report on some developments made in carbon surface pre-treatment and the electrochemical detection scheme which ensure the most sensitive measurement technique is employed. Following pre-treatment of the sensor to ensure homogeneity, DNA probes developed specifically for detection of the KRAS G12D and G13D mutations were immobilized onto low-cost screen printed carbon electrodes using diazonium chemistry and 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride/N-hydroxysuccinimide coupling. Prior to electrochemical detection, the sensor was functionalised with target DNA amplified by standard and specialist PCR methodologies (6.3% increase). Assay development steps and DNA detection experiments were performed using standard voltammetry techniques. Sensitivity (as low as 0.58 ng/μL) and specificity (>300%) was achieved by detecting mutant KRAS G13D PCR amplicons against a background of wild-type KRAS DNA from the representative cancer sample and our findings give rise to the basis of a simple and very low-cost system for measuring ctDNA biomarkers in patient samples. The current time to receive results from the system was 3.5 h with appreciable scope for optimisation, thus far comparing favourably to the UK National Health Service biopsy service where patients can wait for weeks for biopsy results. Full article

(This article belongs to the Special Issue Selected Papers from the 1st International Electronic Conference on Biosensors (IECB 2020))

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12 pages, 3768 KiB

Open AccessArticle

2D Nanomaterial, Ti₃C₂ MXene-Based Sensor to Guide Lung Cancer Therapy and Management

by Mahek Sadiq, Lizhi Pang, Michael Johnson, Venkatachalem Sathish, Qifeng Zhang and Danling Wang

Biosensors 2021, 11(2), 40; https://doi.org/10.3390/bios11020040 - 04 Feb 2021

Cited by 18 | Viewed by 3833

Abstract

Major advances in cancer control can be greatly aided by early diagnosis and effective treatment in its pre-invasive state. Lung cancer (small cell and non-small cell) is a leading cause of cancer-related deaths among both men and women around the world. A lot [...] Read more.

Major advances in cancer control can be greatly aided by early diagnosis and effective treatment in its pre-invasive state. Lung cancer (small cell and non-small cell) is a leading cause of cancer-related deaths among both men and women around the world. A lot of research attention has been directed toward diagnosing and treating lung cancer. A common method of lung cancer treatment is based on COX-2 (cyclooxygenase-2) inhibitors. This is because COX-2 is commonly overexpressed in lung cancer and also the abundance of its enzymatic product prostaglandin E2 (PGE₂). Instead of using traditional COX-2 inhibitors to treat lung cancer, here, we introduce a new anti-cancer strategy recently developed for lung cancer treatment. It adopts more abundant omega-6 (ω-6) fatty acids such as dihomo-γ-linolenic acid (DGLA) in the daily diet and the commonly high levels of COX-2 expressed in lung cancer to promote the formation of 8-hydroxyoctanoic acid (8-HOA) through a new delta-5-desaturase (D5Di) inhibitor. The D5Di does not only limit the metabolic product, PGE_2, but also promote the COX-2 catalyzed DGLA peroxidation to form 8-HOA, a novel anti-cancer free radical byproduct. Therefore, the measurement of the PGE₂ and 8-HOA levels in cancer cells can be an effective method to treat lung cancer by providing in-time guidance. In this paper, we mainly report on a novel sensor, which is based on a newly developed functionalized nanomaterial, 2-dimensional nanosheets, or Ti₃C₂ MXene. The preliminary results have proven to sensitively, selectively, precisely, and effectively detect PGE₂ and 8-HOA in A549 lung cancer cells. The capability of the sensor to detect trace level 8-HOA in A549 has been verified in comparison with the traditional gas chromatography–mass spectrometry (GC–MS) method. The sensing principle could be due to the unique structure and material property of Ti₃C₂ MXene: a multilayered structure and extremely large surface area, metallic conductivity, and ease and versatility in surface modification. All these make the Ti₃C₂ MXene-based sensor selectively adsorb 8-HOA molecules through effective charge transfer and lead to a measurable change in the conductivity of the material with a high signal-to-noise ratio and excellent sensitivity. Full article

(This article belongs to the Special Issue Selected Papers from the 1st International Electronic Conference on Biosensors (IECB 2020))

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16 pages, 4187 KiB

Open AccessArticle

Vibrational Spectra of Nucleotides in the Presence of the Au Cluster Enhancer in MD Simulation of a SERS Sensor

by Tatiana Zolotoukhina, Momoko Yamada and Shingo Iwakura

Biosensors 2021, 11(2), 37; https://doi.org/10.3390/bios11020037 - 29 Jan 2021

Cited by 3 | Viewed by 2152

Abstract

Surface-enhanced Raman scattering (SERS) nanoprobes have shown tremendous potential in in vivo imaging. The development of single oligomer resolution in the SERS promotes experiments on DNA and protein identification using SERS as a nanobiosensor. As Raman scanners rely on a multiple spectrum acquisition, [...] Read more.

Surface-enhanced Raman scattering (SERS) nanoprobes have shown tremendous potential in in vivo imaging. The development of single oligomer resolution in the SERS promotes experiments on DNA and protein identification using SERS as a nanobiosensor. As Raman scanners rely on a multiple spectrum acquisition, faster imaging in real-time is required. SERS weak signal requires averaging of the acquired spectra that erases information on conformation and interaction. To build spectral libraries, the simulation of measurement conditions and conformational variations for the nucleotides relative to enhancer nanostructures would be desirable. In the molecular dynamic (MD) model of a sensing system, we simulate vibrational spectra of the cytosine nucleotide in FF2/FF3 potential in the dynamic interaction with the Au20 nanoparticles (NP) (EAM potential). Fourier transfer of the density of states (DOS) was performed to obtain the spectra of bonds in reaction coordinates for nucleotides at a resolution of 20 to 40 cm⁻¹. The Au₂₀ was optimized by ab initio density functional theory with generalized gradient approximation (DFT GGA) and relaxed by MD. The optimal localization of nucleotide vs. NP was defined and the spectral modes of both components vs. interaction studied. Bond-dependent spectral maps of nucleotide and NP have shown response to interaction. The marker frequencies of the Au₂₀—nucleotide interaction have been evaluated. Full article

(This article belongs to the Special Issue Selected Papers from the 1st International Electronic Conference on Biosensors (IECB 2020))

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15 pages, 2993 KiB

Open AccessEditor’s ChoiceArticle

Porous Silicon Biosensor for the Detection of Bacteria through Their Lysate

by Roselien Vercauteren, Audrey Leprince, Jacques Mahillon and Laurent A. Francis

Biosensors 2021, 11(2), 27; https://doi.org/10.3390/bios11020027 - 20 Jan 2021

Cited by 7 | Viewed by 3446

Abstract

Porous silicon (PSi) has been widely used as a biosensor in recent years due to its large surface area and its optical properties. Most PSi biosensors consist in close-ended porous layers, and, because of the diffusion-limited infiltration of the analyte, they lack sensitivity [...] Read more.

Porous silicon (PSi) has been widely used as a biosensor in recent years due to its large surface area and its optical properties. Most PSi biosensors consist in close-ended porous layers, and, because of the diffusion-limited infiltration of the analyte, they lack sensitivity and speed of response. In order to overcome these shortcomings, PSi membranes (PSiMs) have been fabricated using electrochemical etching and standard microfabrication techniques. In this work, PSiMs have been used for the optical detection of Bacillus cereus lysate. Before detection, the bacteria are selectively lysed by PlyB221, an endolysin encoded by the bacteriophage Deep-Blue targeting B. cereus. The detection relies on the infiltration of bacterial lysate inside the membrane, which induces a shift of the effective optical thickness. The biosensor was able to detect a B. cereus bacterial lysate, with an initial bacteria concentration of 10⁵ colony forming units per mL (CFU/mL), in only 1 h. This proof-of-concept also illustrates the specificity of the lysis before detection. Not only does this detection platform enable the fast detection of bacteria, but the same technique can be extended to other bacteria using selective lysis, as demonstrated by the detection of Staphylococcus epidermidis, selectively lysed by lysostaphin. Full article

(This article belongs to the Special Issue Selected Papers from the 1st International Electronic Conference on Biosensors (IECB 2020))

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18 pages, 2426 KiB

Open AccessArticle

Aptamer–Target–Gold Nanoparticle Conjugates for the Quantification of Fumonisin B1

by Vicente Antonio Mirón-Mérida, Yadira González-Espinosa, Mar Collado-González, Yun Yun Gong, Yuan Guo and Francisco M. Goycoolea

Biosensors 2021, 11(1), 18; https://doi.org/10.3390/bios11010018 - 08 Jan 2021

Cited by 17 | Viewed by 3660

Abstract

Fumonisin B1 (FB1), a mycotoxin classified as group 2B hazard, is of high importance due to its abundance and occurrence in varied crops. Conventional methods for detection are sensitive and selective; however, they also convey disadvantages such as long assay times, expensive equipment [...] Read more.

Fumonisin B1 (FB1), a mycotoxin classified as group 2B hazard, is of high importance due to its abundance and occurrence in varied crops. Conventional methods for detection are sensitive and selective; however, they also convey disadvantages such as long assay times, expensive equipment and instrumentation, complex procedures, sample pretreatment and unfeasibility for on-site analysis. Therefore, there is a need for quick, simple and affordable quantification methods. On that note, aptamers (ssDNA) are a good alternative for designing specific and sensitive biosensing techniques. In this work, the assessment of the performance of two aptamers (40 and 96 nt) on the colorimetric quantification of FB1 was determined by conducting an aptamer–target incubation step, followed by the addition of gold nanoparticles (AuNPs) and NaCl. Although MgCl₂ and Tris-HCl were, respectively, essential for aptamer 96 and 40 nt, the latter was not specific for FB1. Alternatively, the formation of Aptamer (96 nt)–FB1–AuNP conjugates in MgCl₂ exhibited stabilization to NaCl-induced aggregation at increasing FB1 concentrations. The application of asymmetric flow field-flow fractionation (AF4) allowed their size separation and characterization by a multidetection system (UV-VIS, MALS and DLS online), with a reduction in the limit of detection from 0.002 µg/mL to 56 fg/mL. Full article

(This article belongs to the Special Issue Selected Papers from the 1st International Electronic Conference on Biosensors (IECB 2020))

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15 pages, 2831 KiB

Open AccessArticle

A Comparative Study of Approaches to Improve the Sensitivity of Lateral Flow Immunoassay of the Antibiotic Lincomycin

by Kseniya V. Serebrennikova, Olga D. Hendrickson, Elena A. Zvereva, Demid S. Popravko, Anatoly V. Zherdev, Chuanlai Xu and Boris B. Dzantiev

Biosensors 2020, 10(12), 198; https://doi.org/10.3390/bios10120198 - 03 Dec 2020

Cited by 9 | Viewed by 2913

Abstract

This study provides a comparative assessment of the various nanodispersed markers and related detection techniques used in the immunochromatographic detection of an antibiotic lincomycin (LIN). Improving the sensitivity of the competitive lateral flow immunoassay is important, given the increasing demands for the monitoring [...] Read more.

This study provides a comparative assessment of the various nanodispersed markers and related detection techniques used in the immunochromatographic detection of an antibiotic lincomycin (LIN). Improving the sensitivity of the competitive lateral flow immunoassay is important, given the increasing demands for the monitoring of chemical contaminants in food. Gold nanoparticles (AuNPs) and CdSe/ZnS quantum dots (QDs) were used for the development and comparison of three approaches for the lateral flow immunoassay (LFIA) of LIN, namely, colorimetric, fluorescence, and surface-enhanced Raman spectroscopy (SERS)-based LFIAs. It was demonstrated that, for colorimetric and fluorescence analysis, the detection limits were comparable at 0.4 and 0.2 ng/mL, respectively. A SERS-based method allowed achieving the gain of five orders of magnitude in the assay sensitivity (1.4 fg/mL) compared to conventional LFIAs. Therefore, an integration of a SERS reporter into the LFIA is a promising tool for extremely sensitive quantitative detection of target analytes. However, implementation of this time-consuming technique requires expensive equipment and skilled personnel. In contrast, conventional AuNP- and QD-based LFIAs can provide simple, rapid, and inexpensive point-of-care testing for practical use. Full article

(This article belongs to the Special Issue Selected Papers from the 1st International Electronic Conference on Biosensors (IECB 2020))

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13 pages, 4439 KiB

Open AccessArticle

Newly Developed System for the Robust Detection of Listeria monocytogenes Based on a Bioelectric Cell Biosensor

by Agni Hadjilouka, Konstantinos Loizou, Theofylaktos Apostolou, Lazaros Dougiakis, Antonios Inglezakis and Dimitrios Tsaltas

Biosensors 2020, 10(11), 178; https://doi.org/10.3390/bios10110178 - 17 Nov 2020

Cited by 10 | Viewed by 3020

Abstract

Human food-borne diseases caused by pathogenic bacteria have been significantly increased in the last few decades causing numerous deaths worldwide. The standard analyses used for their detection have significant limitations regarding cost, special facilities and equipment, highly trained staff, and a long procedural [...] Read more.

Human food-borne diseases caused by pathogenic bacteria have been significantly increased in the last few decades causing numerous deaths worldwide. The standard analyses used for their detection have significant limitations regarding cost, special facilities and equipment, highly trained staff, and a long procedural time that can be crucial for foodborne pathogens with high hospitalization and mortality rates, such as Listeria monocytogenes. This study aimed to develop a biosensor that could detect L. monocytogenes rapidly and robustly. For this purpose, a cell-based biosensor technology based on the Bioelectric Recognition Assay (BERA) and a portable device developed by EMBIO Diagnostics, called B.EL.D (Bio Electric Diagnostics), were used. Membrane engineering was performed by electroinsertion of Listeria monocytogenes homologous antibodies into the membrane of African green monkey kidney (Vero) cells. The newly developed biosensor was able to detect the pathogen’s presence rapidly (3 min) at concentrations as low as 10² CFU mL⁻¹, demonstrating a higher sensitivity than most existing biosensor-based methods. In addition, lack of cross-reactivity with other Listeria species, as well as with Escherichia coli, was shown, thus, indicating biosensor’s significant specificity against L. monocytogenes. Full article

(This article belongs to the Special Issue Selected Papers from the 1st International Electronic Conference on Biosensors (IECB 2020))

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10 pages, 789 KiB

Open AccessPerspective

Space Biology Research and Biosensor Technologies: Past, Present, and Future

by Ada Kanapskyte, Elizabeth M. Hawkins, Lauren C. Liddell, Shilpa R. Bhardwaj, Diana Gentry and Sergio R. Santa Maria

Biosensors 2021, 11(2), 38; https://doi.org/10.3390/bios11020038 - 29 Jan 2021

Cited by 15 | Viewed by 5556

Abstract

In light of future missions beyond low Earth orbit (LEO) and the potential establishment of bases on the Moon and Mars, the effects of the deep space environment on biology need to be examined in order to develop protective countermeasures. Although many biological [...] Read more.

In light of future missions beyond low Earth orbit (LEO) and the potential establishment of bases on the Moon and Mars, the effects of the deep space environment on biology need to be examined in order to develop protective countermeasures. Although many biological experiments have been performed in space since the 1960s, most have occurred in LEO and for only short periods of time. These LEO missions have studied many biological phenomena in a variety of model organisms, and have utilized a broad range of technologies. However, given the constraints of the deep space environment, upcoming deep space biological missions will be largely limited to microbial organisms and plant seeds using miniaturized technologies. Small satellites such as CubeSats are capable of querying relevant space environments using novel, miniaturized instruments and biosensors. CubeSats also provide a low-cost alternative to larger, more complex missions, and require minimal crew support, if any. Several have been deployed in LEO, but the next iterations of biological CubeSats will travel beyond LEO. They will utilize biosensors that can better elucidate the effects of the space environment on biology, allowing humanity to return safely to deep space, venturing farther than ever before. Full article

(This article belongs to the Special Issue Selected Papers from the 1st International Electronic Conference on Biosensors (IECB 2020))

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