This thesis (carried out at Institute of Chemistry for Molecular Recognition, Italian, National Research Council) focuses on the use of polymers for bioanalytical applications, ranging from microarray technology, capillary electrophoresis, DNA sequencing, DNA conformational changes observation. Because of such a variety of applications, this work has been divided into two parts. Part one mainly deals with surface modification by means of polymeric coatings. Our reasearch group boasts a deep knowledge in solid support derivatization and on the wake of this expertise an already introduced polymer has been deeply characterized and new ones proposed. • Chapter 1 provides a short and non-exhaustive review on surface modification methodologies and their uses to control surface properties. Control of surface chemical-physical properties is of utmost importance in order to develop microanalytical devices. One of the main purposes is the immobilization of biomolecules on solid supports mantaining their native structure and avoiding aspecific bonding. Polymers have revealed to be one of the best choice to achieve these aims and severasl techniques have been explored and here described. • Chapter 2 introduces microarray technology, one of the most important tenchnique to perform high-throughput biorecognition analysis, thanks to its multiplexing and miniaturization features. In 2004, our group has introduced a novel and easy-to-use polymeric coating, which was lacking of a proper characterization. Here we report the optimization of its synthesis protocol and a deep characterization regarding its molecular mass, stabilty, swelling and functionality. • Chapter 3 introduces adsorbed stable polymeric coatings for Capillary Electrophoresis, one of the most powerful tool for protein detection and separation. This technique requires a perfect control of surface properties to avoid protein interaction with capillary walls and to regulate electroosmotic flow, which would affect the separation. Here we describe a new employment of the coating already mentioned in Chapter 1 and we introduce a new one to prevent protein interactions with capillary surface and electroosmotic flow regulation. • Chapter 4 describes the design, the synthesis and the characterization of new smart coatings to generate a platform for DNA switch bionsensor who found an interest application in the study of DNA confomational chnages introduced by protein binding. This work has been carried out in collaboration with Prof. Selim M. Unlu from Boston University, USA. Part two basically consists in the last chapter of this thesis: • Chapter 5 describes the synthesis and the characterization of ultra-high molar mass linear polymeric matrices for DNA sequencing in capillary electrophoresis. The synthetic process we propose is an inverse emulsion polymerization, which presents several advantages respect to the classic random radical polymerization performed in free solution. We optimized the emulsion formulation and the entire synthetic method and we obtained a scalable process that produces linear polyacrylamide with molecular mass in the order of 20 MDa. Its performance has then been evaluated by sequencing DNA standards in capillary electrophoresis. This work has been carried out in collaboration with Prof. Anneliese E. Barron from Stanford University, CA, USA
DESIGN, SYNTHESIS AND CHARACTERIZATION OF NEW POLYMERS FOR BIOANALYTICAL APPLICATIONS / L. Sola ; tutor: C. De Micheli ; co-tutor: M. Chiari. - Milano : Università degli studi di Milano. Universita' degli Studi di Milano, 2012 Feb 14. ((24. ciclo, Anno Accademico 2011.
|Titolo:||DESIGN, SYNTHESIS AND CHARACTERIZATION OF NEW POLYMERS FOR BIOANALYTICAL APPLICATIONS|
|Data di pubblicazione:||14-feb-2012|
|Parole Chiave:||microarray ; capillary electrophoresis ; DNA sequencing ; DNA switching ; Immobilines ; polymer ; coating|
|Settore Scientifico Disciplinare:||Settore CHIM/08 - Chimica Farmaceutica|
|Citazione:||DESIGN, SYNTHESIS AND CHARACTERIZATION OF NEW POLYMERS FOR BIOANALYTICAL APPLICATIONS / L. Sola ; tutor: C. De Micheli ; co-tutor: M. Chiari. - Milano : Università degli studi di Milano. Universita' degli Studi di Milano, 2012 Feb 14. ((24. ciclo, Anno Accademico 2011.|
|Appare nelle tipologie:||Tesi di dottorato|