Optical enzyme-based sensors surfaces

Arnold et al. (1987) described an optoelectronic ethanol sensor based on fluorimetric detection of NADH formed in the reaction catalyzed by ADH. The enzyme was fixed to the inner surface of a membrane permeable to volatile substances, which separated the sample from the internal sensor solution. This solution contained NADH and semicarbazide, so that no reagent had to be added to the sample. The arrangement was named an internal optical enzyme sensor . 

Electrochemical sensor fabrication has dominated the analytical application of polymers. In some sensors the polymer film acts as a membrane for the preconcentration of ions or elements before electrochemical detection. Polymers also serve as materials for electrode modification that lower the potential for detecting analytes. In addition, some polymer films function as electrocatalytic surfaces. Using a polymer in biosensors is a very rapidly developing area of electroanalytical chemistry. Polymeric matrix modifiers have been applied as diffusional barriers in constructing not only sensitive amperometric biosensors, but also electrochemical sensors that apply potentiometric, conductimetric, optical, and gas-sensing transducer systems. The principles, operations, and application of potentiometric, conductimetric, optical and gas sensors are described in Refs. 13, 39-41. In this chapter, we focus mainly on amperometric biosensors based on redox enzymes. 

Dancil KPS, Greiner DP, Sailor MJ (1999) A porous silicon optical biosensor detection of reversible binding of IgG to a protein A-modified surface. J Am Chem Soc 121 7925 De Stefano L, Arcari P, Lambert A, Sanges C, Rotiroti L, Rea I, Rendina I (2007) DNA optical detection based on porous silicon technology from biosensors to biochips. Sensors 7 214 DeLouise LA, Kou PM, Miller BL (2005) Cross-correlation of optical microcavity biosensor response with immobilized enzyme activity. Insights into biosensor sensitivity. Anal Chem 77 3222... 

Finally, in optical-based sensors and assays, there is the development of label-free bioanalytic detection on prepared membrane surfaces, ultra sensitive detection of microbial agents, the development of fiber-optic based enzyme sensors, multi-analyte breast cancer immunoassays, and photodynamic therapy of osteosarcoma in veterinary patients. 

In this part we will describe recent achievements in the development of biosensors based on DNA/RNA aptamers. These biosensors are usually prepared by immobilization of aptamer onto a solid support by various methods using chemisorption (aptamer is modified by thiol group) or by avidin-biotin technology (aptamer is modified by biotin) or by covalent attachment of amino group-labeled aptamer to a surface of self-assembly monolayer of 11-mercaptoundecanoic acid (11-MUA). Apart from the method of aptamer immobilization, the biosensors differ in the signal generation. To date, most extensively studied were the biosensors based on optical methods (fluorescence, SPR) and acoustic sensors based mostly on thickness shear mode (TSM) method. However, recently several investigators reported electrochemical sensors based on enzyme-labeled aptamers, electrochemical indicators and impedance spectroscopy methods of detection. 

A DNA optical sensor system was proposed by Cass and co-workers [35] based on the combination of sandwich solution hybridization, magnetic bead capture, flow injection and chemiluminescence for the rapid detection of DNA hybridization. Sandwich solution hybridization uses two sets of DNA probes, one labelled with biotin, the other with an enzyme marker and hybridization is performed in solution where the mobility is greater and the hybridization process is faster, rather than on a surface. The hybrids were bound to the streptavidin-coated magnetic beads through biotin-streptavidin binding reaction. A chemiluminescence fibre-optic biosensor for the detection of hybridization of horseradish peroxidase-labelled complementary DNA to covalent immobilized DNA probes was developed by Zhou and co-workers [36]. 

Enzyme DNA hybridization assays with electrochemical detection can offer enhanced sensitivity and reduced instrumentation costs in comparison with their optical counterparts. Efforts to prevent non-specific binding of the codissolved enzyme and to avoid fouling problems by selecting conditions suitable to amplify the electrode response have been reported by Heller and co-workers [107]. A disposable electrochemical sensor based on an ion-exchange film-coated screen-printed electrode was described by Limoges and co-workers for an enzyme nucleic acid hybridization assay using alkaline phosphatase [108] or horseradish peroxidase [109]. In another methodology to improve sensitivity, a carbon paste electrode with an immobilized nucleotide on the electrode surface and methylene blue as hybridization indicator was coupled, by Mascini and co-workers [110], with PGR amplification of DNA extracted from human blood for the electrochemical detection of virus. 

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