Biosensor electrode fabrication techniques

In recent years, there are more applications based on the layer-by-layer fabrication techniques for CNT-modified electrodes. This technique clearly provides thinner and more isolated CNTs compared with other methods such as CNT-composite and CNT coated electrodes in which CNTs are in the form of big bundles. This method should help biomolecules such as enzymes and DNA to interact more effectively with CNTs than other methods, and sensors based on this technique are expected to be more sensitive. Important biosensors such as glucose sensors have been developed using this technique, and further development of other sensors based on the layer-by-layer technique is expected. 

Karube and Yokoyama presented an overview on the developments in the biosensor technology [60]. The overview describes the use of micromachining fabrication techniques for the construction of detection units for FIA, electrochemical flow cells and chemiluminescence detectors. Acetylcholine microsensors using carbon fiber electrodes and glutamate microsensors for neuroscience were discussed. 

On the other hand, organic dyes in combination with CNTs are also used in electrode fabrication and hybridization techniques for developing an ultrasensitive, selective, and miniaturized electrochemical DNA biosensor for quick and reliable DNA sequence analysis. CNTs have good biocompatibility which allows them to increase the attached DNA amount on the substrate surface, thanks to their high surface area, an enhanced electronic conductivity, and a high mechanical resistance in electroanalytical chemistry can accelerate the electron-transfer rate between the redox active ssDNA molecule and electrode. 

The packaging (i.e., electrical insulation for operation in electrolytes) is more difficult with SAWs due to their rectangular geometry. SAWs are easier to fabricate with lithographic microfabrication techniques and therefore are more suitable for use in an array (Ricco et al., 1998). The choice of electrode materials is critical for QCM, where acoustic impedance mismatch can result in substantial lowering of the Q factor of the device. On the other hand, it does not play any role in the SAW devices. The energy losses to the condensed medium are higher in SAWs and this fact makes them even less suitable for operation in liquids. Nevertheless, SAW biosensors have been reported (Marx, 2003). 

Layer-by-layer (LbL) assembly is a unique technique for the fabrication of composite films with precise thickness control at the nanometer scale [111, 112], The method is based on the alternate adsorption of oppositely charged species from their solutions. The attractive feature of this approach is its ability to assemble complex structures from modular components, and integrate them into self-assembling constructions for a wide range of applications. The LbL method has been successfully exploited in the construction of dendrimer biosensors [113,114], The LbL films provide a favorable environment for the intimate contact between the dendrimer and biomolecule (enzymes or proteins), promoting a direct electron transfer between them and the underlying electrodes. 

In this chapter, we review the recent progress in the development of different metal oxide nanoparticles with various shapes and size for fabrication of biosensors. The development of metal oxide nanomaterials surface film for direct electron exchange between electrodes and redox enzymes and proteins will be summarizing. The electrochemical properties, stability and biocatalytic activity of the proposed biosensors will be discussed. The biocompatibility of the metal oxide nanomaterials for enzymes and biomolecules will be evaluated. We will briefly describe some techniques for the investigation of proteins and enzymes when adsorbed to the electrode surfaces. Cyclic voltammetry, impedance spectroscopy, UV-visible spectroscopy and surface imaging techniques were used for surface characterization and bioactivity measuring. 

Due to high biocompability and large surface are of cobalt oxide nanoparticles it can be used for immobilization of other biomolecules. Flavin adenine FAD is a flavoprotein coenzyme that plays an important biological role in many oxidoreductase processes and biochemical reactions. The immobilized FAD onto different electrode surfaces provides a basis for fabrication of sensors, biosensors, enzymatic reactors and biomedical devices. The electrocatalytic oxidation of NADH on the surface of graphite electrode modified with immobilization of FAD was investigated [276], Recently we used cyclic voltammetry as simple technique for cobalt-oxide nanoparticles formation and immobilization flavin adenine dinucleotide (FAD) [277], Repeated cyclic voltammograms of GC/ CoOx nanoparticles modified electrode in buffer solution containing FAD is shown in Fig.37A. 

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