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Biosensors, Volume 2, Issue 3 (September 2012) – 7 articles , Pages 245-340

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Open AccessArticle
Development of a Fish Cell Biosensor System for Genotoxicity Detection Based on DNA Damage-Induced Trans-Activation of p21 Gene Expression
Biosensors 2012, 2(3), 318-340; https://doi.org/10.3390/bios2030318 - 10 Sep 2012
Cited by 10 | Viewed by 3731
Abstract
p21CIP1/WAF1 is a p53-target gene in response to cellular DNA damage. Here we report the development of a fish cell biosensor system for high throughput genotoxicity detection of new drugs, by stably integrating two reporter plasmids of pGL3-p21-luc (human [...] Read more.
p21CIP1/WAF1 is a p53-target gene in response to cellular DNA damage. Here we report the development of a fish cell biosensor system for high throughput genotoxicity detection of new drugs, by stably integrating two reporter plasmids of pGL3-p21-luc (human p21 promoter linked to firefly luciferase) and pRL-CMV-luc (CMV promoter linked to Renilla luciferase) into marine flatfish flounder gill (FG) cells, referred to as p21FGLuc. Initial validation of this genotoxicity biosensor system showed that p21FGLuc cells had a wild-type p53 signaling pathway and responded positively to the challenge of both directly acting genotoxic agents (bleomycin and mitomycin C) and indirectly acting genotoxic agents (cyclophosphamide with metabolic activation), but negatively to cyclophosphamide without metabolic activation and the non-genotoxic agents ethanol and D-mannitol, thus confirming a high specificity and sensitivity, fast and stable response to genotoxic agents for this easily maintained fish cell biosensor system. This system was especially useful in the genotoxicity detection of Di(2-ethylhexyl) phthalate (DEHP), a rodent carcinogen, but negatively reported in most non-mammalian in vitro mutation assays, by providing a strong indication of genotoxicity for DEHP. A limitation for this biosensor system was that it might give false positive results in response to sodium butyrate and any other agents, which can trans-activate the p21 gene in a p53-independent manner. Full article
(This article belongs to the Special Issue Nano and Micro DNA/RNA Sensors)
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Open AccessArticle
Functional Conducting Polymers via Thiol-ene Chemistry
Biosensors 2012, 2(3), 305-317; https://doi.org/10.3390/bios2030305 - 31 Aug 2012
Cited by 15 | Viewed by 4468
Abstract
We demonstrate here that thiol-ene chemistry can be used to provide side-chain functionalized monomers based on 3,4-propylenedioxythiophene (ProDOT) containing ionic, neutral, hydrophobic, and hydrophilic side chains. All reactions gave high yields and purification could generally be accomplished through precipitation. These monomers were polymerized [...] Read more.
We demonstrate here that thiol-ene chemistry can be used to provide side-chain functionalized monomers based on 3,4-propylenedioxythiophene (ProDOT) containing ionic, neutral, hydrophobic, and hydrophilic side chains. All reactions gave high yields and purification could generally be accomplished through precipitation. These monomers were polymerized either chemically or electro-chemically to give soluble materials or conductive films, respectively. This strategy provides for facile tuning of the solubility, film surface chemistry, and film morphology of this class of conducting polymers. Full article
(This article belongs to the Special Issue Organic Electronic Bio-Devices)
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Open AccessArticle
Biomolecular Interaction Analysis of Gestrinone-anti-Gestrinone Using Arrays of High Aspect Ratio SU-8 Nanopillars
Biosensors 2012, 2(3), 291-304; https://doi.org/10.3390/bios2030291 - 14 Aug 2012
Cited by 8 | Viewed by 3893
Abstract
In this paper, label-free biosensing for antibody screening by periodic lattices of high-aspect ratio SU-8 nano-pillars (BICELLs) is presented. As a demonstration, the determination of anti-gestrinone antibodies from whole rabbit serum is carried out, and for the first time, the dissociation constant (K [...] Read more.
In this paper, label-free biosensing for antibody screening by periodic lattices of high-aspect ratio SU-8 nano-pillars (BICELLs) is presented. As a demonstration, the determination of anti-gestrinone antibodies from whole rabbit serum is carried out, and for the first time, the dissociation constant (KD = 6 nM) of antigen-antibody recognition process is calculated using this sensing system. After gestrinone antigen immobilization on the BICELLs, the immunorecognition was performed. The cells were interrogated vertically by using micron spot size Fourier transform visible and IR spectrometry (FT-VIS-IR), and the dip wavenumber shift was monitored. The biosensing assay exhibited good reproducibility and sensitivity (LOD = 0.75 ng/mL). Full article
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Open AccessArticle
A Comparative Study of Impedance versus Optical Label-Free Systems Relative to Labelled Assays in a Predominantly Gi Coupled GPCR (C5aR) Signalling
Biosensors 2012, 2(3), 273-290; https://doi.org/10.3390/bios2030273 - 26 Jul 2012
Cited by 12 | Viewed by 4344
Abstract
Profiling ligand function on G-protein coupled receptors (GPCRs) typically involves using transfected cells over-expressing a target of interest, a labelled ligand, and intracellular secondary messenger reporters. In contrast, label-free assays are sensitive enough to allow detection in native cells, which may provide a [...] Read more.
Profiling ligand function on G-protein coupled receptors (GPCRs) typically involves using transfected cells over-expressing a target of interest, a labelled ligand, and intracellular secondary messenger reporters. In contrast, label-free assays are sensitive enough to allow detection in native cells, which may provide a more physiologically relevant readout. Here, we compare four agonists (native agonists, a peptide full agonist and a peptide partial agonist) that stimulate the human inflammatory GPCR C5aR. The receptor was challenged when present in human monocyte-derived macrophages (HMDM) versus stably transfected human C5aR-CHO cells. Receptor activation was compared on label-free optical and impedance biosensors and contrasted with results from two traditional reporter assays. The rank order of potencies observed across label-free and pathway specific assays was similar. However, label-free read outs gave consistently lower potency values in both native and transfected cells. Relative to pathway-specific assays, these technologies measure whole-cell responses that may encompass multiple signalling events, including down-regulatory events, which may explain the potency discrepancies observed. These observations have important implications for screening compound libraries against GPCR targets and for selecting drug candidates for in vivo assays. Full article
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Open AccessEditorial
Electrochemical Based Biosensors
Biosensors 2012, 2(3), 269-272; https://doi.org/10.3390/bios2030269 - 24 Jul 2012
Cited by 9 | Viewed by 3455
Abstract
This editorial summarizes the general approaches of the electrochemical based biosensors described in the manuscripts published in this Special Issue. Electrochemical based biosensors are scientifically and economically important for the detection and early diagnosis of many diseases, and they will be increasing used [...] Read more.
This editorial summarizes the general approaches of the electrochemical based biosensors described in the manuscripts published in this Special Issue. Electrochemical based biosensors are scientifically and economically important for the detection and early diagnosis of many diseases, and they will be increasing used and developed in the coming years. The importance of the selection of recognition processes, fabrication techniques and biosensor materials will be introduced. Full article
(This article belongs to the Special Issue Electrochemical Based Biosensors)
Open AccessArticle
Effect of Toxic Components on Microbial Fuel Cell-Polarization Curves and Estimation of the Type of Toxic Inhibition
Biosensors 2012, 2(3), 255-268; https://doi.org/10.3390/bios2030255 - 11 Jul 2012
Cited by 15 | Viewed by 4901
Abstract
Polarization curves are of paramount importance for the detection of toxic components in microbial fuel cell (MFC) based biosensors. In this study, polarization curves were made under non-toxic conditions and under toxic conditions after the addition of various concentrations of nickel, bentazon, sodiumdodecyl [...] Read more.
Polarization curves are of paramount importance for the detection of toxic components in microbial fuel cell (MFC) based biosensors. In this study, polarization curves were made under non-toxic conditions and under toxic conditions after the addition of various concentrations of nickel, bentazon, sodiumdodecyl sulfate and potassium ferricyanide. The experimental polarization curves show that toxic components have an effect on the electrochemically active bacteria in the cell. (Extended) Butler Volmer Monod (BVM) models were used to describe the polarization curves of the MFC under nontoxic and toxic conditions. It was possible to properly fit the (extended) BVM models using linear regression techniques to the polarization curves and to distinguish between different types of kinetic inhibitions. For each of the toxic components, the value of the kinetic inhibition constant Ki was also estimated from the experimental data. The value of Ki indicates the sensitivity of the sensor for a specific component and thus can be used for the selection of the biosensor for a toxic component. Full article
(This article belongs to the Special Issue Electrochemical Based Biosensors)
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Open AccessArticle
Thiol- and Biotin-Labeled Probes for Oligonucleotide Quartz Crystal Microbalance Biosensors of Microalga Alexandrium Minutum
Biosensors 2012, 2(3), 245-254; https://doi.org/10.3390/bios2030245 - 04 Jul 2012
Cited by 6 | Viewed by 3615
Abstract
Two quartz crystal microbalance oligonucleotide biosensors of a toxic microalga gene sequence (Alexandrium Minutum) have been designed. Grafting on a gold surface of 20-base thiol- or biotin-labeled probe, and selective hybridization with the complementary 20-base target, have been monitored in situ [...] Read more.
Two quartz crystal microbalance oligonucleotide biosensors of a toxic microalga gene sequence (Alexandrium Minutum) have been designed. Grafting on a gold surface of 20-base thiol- or biotin-labeled probe, and selective hybridization with the complementary 20-base target, have been monitored in situ with a 27 MHz quartz crystal microbalance under controlled hydrodynamic conditions. The frequency of the set up is stable to within a few hertz, corresponding to the nanogram scale, for three hour experiments. DNA recognition by the two biosensors is efficient and selective. Hybridization kinetic curves indicate that the biosensor designed with the thiol-labeled probe is more sensitive, and that the biosensor designed with the biotin-labeled probe has a shorter time response and a higher hybridization efficiency. Full article
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