Hext -- Chapter Ellington -- Chapter Turner -- Chapter Rickus, Bruce Dunn, and Jeffrey I. Zink -- Chapter Membrane-based Biosensors -- Bruce Corell -- Chapter Philbert, -- Jonathan W. Miller and Ron Tjalkens -- Chapter Kenneth Kuno, Ellen R. Goldman, -- George P.
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OPTICAL BIOSENSOR DEVICES AS EARLY DETECTORS OF BIOLOGICAL AND CHEMICAL WARFARE AGENTS
Croddy, C. Perez-Armendariz and J. Hart, Eds. Iqbal, M. Mayo, J. Bruno, B. Bronk, C. Batt and J. Rowe-Taitt, J. Hazzard, K.
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Biosensor - Wikipedia
Piliarik and J. Homola, Data analysis for optical sensors based on spectroscopy of surface plasmons, Meas. Kooyman and L. Current cancer biomarkers are based on overexpressed cancer proteins in blood; however, their number and clinical use are rather limited and require a large population of cancer patients with well-defined clinical staging and outcomes. Analysis of the epigenetic pathways may be more informative, specific, and accurate than the analysis of such protein biomarkers, by the determination of not only the cancer itself, but also the underlying mechanisms by which it is generated.
Recent studies indicate that these regulation pathways participate in collaborative activities resulting in a common outcome. On top of this, it will contribute to a more efficient management of cancer patients, providing an early diagnosis, determining precise tumour staging, and monitoring of treatment. The revolutionary epigenetic inheritance has changed how we understand and deal with cancer.
The dynamic nature and potential reversibility of the epigenetic mechanisms mean that they are appealing therapeutic targets in cancer treatment. Applying therapies focussed on reversion of the altered processes to their normal state would abate the cancer progression in a less invasive and more efficient manner than standard chemotherapies.
Currently, various compounds that can rearrange DNA methylation and histone acetylation patterns are being examined in clinical settings in combination with other drugs.
Developments in real-time polymerase chain reaction PCR have allowed results to be obtained in a matter of a few minutes regarding the methylation status or mRNA isoform shift. Besides, all cancers involve more than one epigenetic mechanism, making the simultaneous detection of multiple epigenetic biomarkers essential.
Emerging trends in diagnostics have promoted the development of diagnostic tools with improved sensitivity and short operation times, such as biosensors. Biosensors can be designed to provide quantitative analytical information with elevated accuracy in a few minutes, using low sample volumes and minimum sample pretreatment.
By definition, a biosensor is a self-contained analytical device that incorporates a biologically active material in intimate contact with an appropriate transduction element for the purpose of detecting, in a very selective way, the concentration or activity of chemical species in any type of sample Figure 1. Figure 1: Schematic diagram of a biosensor device. Adapted with permission from Carrascosa et al. Biosensors are threatening to radically alter our present concept of clinical analysis, beginning many years ago 27 with the introduction of the glucose biosensor signifying a breakthrough in healthcare by the decentralisation and simplicity of analysis at home by one blood-drop.
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In the last decades, a particular interest has been focussed on the development of novel, label-free optical biosensors able to generate a signal directly by the interaction of the analyte of interest with the recognition element, without requiring additional interactions with other probes carrying a label that provides the signal. In general, label-free methods offer potential advantages in terms of simplicity and velocity of the bioanalysis, which may not require washing steps or additional reagents. These biosensors enable the real-time monitoring of the biomolecular interaction, speeding up detection and giving access to the kinetic parameters of the recognition process.
These characteristics allow for the fabrication of smaller, cheaper, and easy-to-use biosensor devices that can accelerate the real implementation of lab-on-a-chip devices in clinical practice. Table 1: Challenges in the biosensing of epigenetic mechanisms and different biosensor approaches.
For instance, the modification of the DNA sequence by methyl-groups has been of interest in many optical biosensor applications. This type of biosensor uses a gold layer as both a surface to immobilise specific probes and a transducer of the signals produced by the refractive index changes at the surface. Specific recognition of methyl-sites and derivatives has been solved with specific antibodies 37,38 or proteins 39 being able to quantify the number of cytosines without the necessity of sample manipulation. In this way, new microfluidic designs have been proposed in order to provide smaller and more easy-to-handle equipment for DNA methylation analyses.
In the case of micro-RNA detection, due to their small size, short RNA regulators are difficult to amplify through conventional methods. In addition, they usually belong to a micro-RNA family with very similar sequences that can distort the analysis with false positive signals. The main objective towards micro-RNA detection relies on two premises: specificity and a high sensitivity to cover a wide range of concentrations.
The ability to perform the experiments directly from an untreated biofluid without the need for purification steps, thereby risking sample input, is also important. To achieve the required sensitivity levels without PCR-amplification steps, researchers used a signal enhancer based on a specific antibody. On the other hand, our team has designed a nanotechnology-based optical biosensor with multiplexing capabilities that has been able to cover the total analytical range from attomolar to nanomolar concentrations, skipping any further enhancement. Its miniaturised size allows for multiplex arrays formats, incorporating 20 nanosensors within the same sensor chip 10 mm width, 31 mm length.
Few optical biosensors have been devised in order to use the alternative splicing regulation route for diagnostic purposes, probably due to the long RNA sequences and the similarity between mRNA isoforms that critically complicate the differentiation between the isoforms. Single mRNA-spliced variants have been identified and quantified in living cells by quantitative imaging.