Sviluppo di biosensori: modifiche di superfici elettrodiche e sistemi di immobilizzazione enzimatica

An amperometric glucose biosensor was developed using an anionic clay matrix (LDH) as enzyme support. The enzyme glucose oxidase (GOx) was immobilized on a layered double hydroxide Ni/Al-NO3 LDH during the electrosynthesis, which was followed by crosslinking with glutaraldehyde (GA) vapours or with GA and bovine serum albumin (GABSA) to avoid the enzyme release. The electrochemical reaction was carried out potentiostatically, at -0.9V vs. SCE, using a rotating disc Pt electrode to assure homogeneity of the electrodeposition suspension, containing GOx, Ni(NO3)2 and Al(NO3)3 in 0.3 M KNO3. The mechanism responsible of the LDH electrodeposition involves the precipitation of the LDH due to the increase of pH at the surface of the electrode, following the cathodic reduction of nitrates. The Pt surface modified with the Ni/Al-NO3 LDH shows a much reduced noise, giving rise to a better signal to noise ratio for the currents relative to H2O2 oxidation, and a linear range for H2O2 determination wider than the one observed for bare Pt electrodes. We pointed out the performances of the biosensor in terms of sensitivity to glucose, calculated from the slope of the linear part of the calibration curve for enzimatically produced H2O2; the sensitivity was dependent on parameters related to the electrodeposition in addition to working conditions. In order to optimise the glucose biosensor performances, with a reduced number of experimental runs, we applied an experimental design. A first screening was performed considering the following variables: deposition time (30 - 120 s), enzyme concentration (0.5 - 3.0 mg/mL), Ni/Al molar ratio (3:1 or 2:1) of the electrodeposition solution at a total metals concentration of 0.03 M and pH of the working buffer solution (5.5-7.0). On the basis of the results from this screening, a full factorial design was carried out, taking into account only enzyme concentration and Ni/Al molar ratio of the electrosynthesis solution. A full factorial design was performed to study linear interactions between factors and their quadratic effects and the optimal setup was evaluated by the isoresponse curves. The significant factors were: enzyme concentration (linear and quadratic terms) and the interaction between enzyme concentration and Ni/Al molar ratio. Since the major obstacle for application of amperometric glucose biosensors is the interference signal resulting from other electro-oxidizable species present in the real matrices, such as ascorbate (AA), the use of different permselective membranes on Pt-LDHGOx modified electrode was discussed with the aim of improving biosensor selectivity and stability. Conventional membranes obtained using Nafion, glutaraldehyde (GA) vapours, GA-BSA were tested together with more innovative materials like palladium hexacyanoferrate (PdHCF) and titania hydrogels. Particular attention has been devoted to hydrogels, because they possess some attractive features, which are generally considered to favour biosensor materials biocompatibility and, consequently, the functional enzyme stability. The Pt-LDH-GOx-PdHCF hydrogel biosensor presented an anti-interferant ability so that to be applied for an accurate glucose analysis in blood. To further improve the biosensor selectivity, protective membranes containing horseradish peroxidase (HRP) were also investigated with the aim of oxidising the interferants before they reach the electrode surface. In such a case glucose determination was also accomplished in real matrices with high AA content. Furthermore, the application of a LDH containing nickel in the oxidised state was performed not only as a support for the enzyme, but also as anti-interferant sistem. The result is very promising and it could be the starting point for further applications in the field of amperometric biosensors; the study could be extended to other oxidase enzymes.

L'idea di metafora nell'opera di Oswald Mathias Ungers

This research focuses on the definition of the complex relationship that exists between theory and project, which - in the architectural work by Oswald Mathias Ungers - is based on several essays and on the publications that - though they have never been collected in an organic text - make up an articulated corpus, so that it is possible to consider it as the foundations of a theory. More specifically, this thesis deals with the role of metaphor in Unger’s theory and its subsequent practical application to his projects. The path leading from theoretical analysis to architectural project is in Ungers’ view a slow and mediated path, where theory is an instrument without which it would not be possible to create the project's foundations. The metaphor is a figure of speech taken from disciplines such as philosophy, aesthetics, linguistics. Using a metaphor implies a transfer of meaning, as it is essentially based on the replacement of a real object with a figurative one. The research is articulated in three parts, each of them corresponding to a text by Ungers that is considered as crucial to understand the development of his architectural thinking. Each text marks three decades of Ungers’ work: the sixties, seventies and eighties. The first part of the research deals with the topic of Großform expressed by Ungers in his publication of 1966 Grossformen im Wohnungsbau, where he defines four criteria based on which architecture identifies with a Großform. One of the hypothesis underlying this study is that there is a relationship between the notion of Großform and the figure of metaphor. The second part of the thesis analyzes the time between the end of the sixties and the seventies, i.e. the time during which Ungers lived in the USA and taught at the Cornell University of Ithaca. The analysis focuses on the text Entwerfen und Denken in Vorstellungen, Metaphern und Analogien, written by Ungers in 1976, for the exhibition MAN transFORMS organized in the Cooper - Hewitt Museum in New York. This text, through which Ungers creates a sort of vocabulary to explain the notions of metaphor, analogy, signs, symbols and allegories, can be defined as the Manifesto of his architectural theory, the latter being strictly intertwined with the metaphor as a design instrument and which is best expressed when he introduces the 11 thesis with P. Koolhaas, P. Riemann, H. Kollhoff and A. Ovaska in Die Stadt in der Stadt in 1977. Berlin das grüne Stadtarchipel. The third part analyzes the indissoluble tie between the use of metaphor and the choice of the topic on which the project is based and, starting from Ungers’ publication in 1982 Architecture as theme, the relationship between idea/theme and image/metaphor is explained. Playing with shapes requires metaphoric thinking, i.e. taking references to create new ideas from the world of shapes and not just from architecture. The metaphor as a tool to interpret reality becomes for Ungers an inquiry method that precedes a project and makes it possible to define the theme on which the project will be based. In Ungers’ case, the architecture of ideas matches the idea of architecture; for Ungers the notions of idea and theme, image and metaphor cannot be separated from each other, the text on thematization of architecture is not a report of his projects, but it represents the need to put them in order and highlight the theme on which they are based.