Organic-inorganic Interfaces for a New Generation of Hybrid Biosensors

Biosensors have by far moved from laboratories benches to the point of use, and, in some cases, their represent technical standards and commercial successes in applications of social interest, such as medical diagnostic or environmental monitoring. Based on biological molecules, but also on their bio-inspired synthetic counterparts, biosensors employ different transducers (optical, potentiometric, volt-amperometric, colorimetric, and so on) converting the molecular interaction information into a measurable electric signal. As the result of a real multi-disciplinary field of science and technology, biosensors can take advantage from each improvement and progress coming from other disciplines: new features and better performances have been reached in the last year due to simplified fabrication methodologies, deep integration of optical or electrical transducers, and, last but not least, microfluidic circuits. More recently, nanostructured components dramatically increased biosensors reliability especially in public health and environmental monitoring. Nevertheless, there is still a pressing demand of innovations which could lead to smaller, faster, and cheaper biosensors systems with ability to provide not only accurate information but also feedback actions to the real world. The fabrication of a new generation of hybrid biodevices, where biological, or bio-inspired, molecules are fully integrated with a micro or a nano technological platform, strongly depends on the bio-compatibilization treatments of the devices surfaces. The design and the realization of bio/non-bio interfaces with specific properties, such as chemical stability, wettability, and biomolecules immobilization ability, are key features in the miniaturization and optimization processes of biosensors. In particular, protein immobilization is a hot topic in biotechnology since commercial solutions, as in the case of DNA microarrays, are not still available. Proteins are, due to their composition, a class of very heterogeneous macromolecules with variable properties. For these reasons, it is extremely difficult to find a common surface suitable for different proteins with a broad range in molecular weight and physical-chemical properties such as charge and hydrophobicity. A further aspect is the orientation of the bound proteins, that could be of crucial relevance for quantitative analysis, interaction studies, and enzymatic reactions. Many different surfaces, and chemical treatments of these surfaces, have been investigated in the last years, but an universal solution for all the applications aforementioned could not be identified. Following this very actual theme, our main focus in this chapter is to discuss different applications in biosensing of a special class of amphiphilic proteins: the hydrophobins. These proteins self-assemble in a nanometric biofilm at the interfaces between water and air, or on the surfaces covered by water solution. New functionalities can be added to the biosensors surfaces without using any chemical or physical treatment, just covering them by a self-assembled protein biofilm. The main topics covered in the following paragraphs are: origin and properties of the hydrophobins; deposition methods of the hydrophobins biofilm on different surfaces and the characterizations techniques we use to determine the physical properties of these bio-interfaces; the features exhibited by the hydrophobins covered surfaces, and finally, the biosensors systems based on hydrophobins biofilms. We outline in this chapter how the peculiarities of these proteins can be of interest in the technological field, beyond their large utilization in biotechnology, nowadays at industrial level. Moreover, the experience matured on this subject can be the paradigm of a new kind of approach in design and realization of the next generation of biosensors.