Polimeri "imprinted" per l'analisi proteomica - Molecularly imprinted Polymers for proteome analysis

The research work carried out in the last three years and presented in this thesis, has been conduced in the Laboratory of Biochemical Methodologies of the Scientific and Technologic Department of the University of Verona, in collaboration with Cranfield Health of the University of Cranfield (UK). One of the main problems in proteome analysis is the presence of high abundance proteins from house-keeping or structural genes, that mask the signal of low abundance ones. The low ones are often potential biomarkers to study specific diseases. In literature different approaches for the removal of high abundance proteins are described, among which electrophoretic and chromatographic pre-fractionation techniques and antibodymediated depletion of specific proteins from biological fluids (i.e., human serum, cerebrospinal fluid). The aim of my PhD thesis has been using Molecularly Imprinted Polymers (MIPs) to manufacture a polymeric synthetic receptor capable of recognising Human Serum Albumin from human serum (60% of the total serum proteins). Molecular imprinting can be defined as the process of template-induced formation of specific recognition sites (binding or catalytic) in a material, where the template drives the positioning and orientation of the material’s structural components by a self-assembling mechanism. The polymer are very easy to prepare and to use, finally cheaper than some methods usually applied. Such a MIP would then be used to remove albumin from serum samples, in order to improve and to help the analysis of the “hidden” proteome. The objectives of my PhD project can be grouped in two different sections, according to which the present thesis is structured: Section I: Development of a new methodology for imprinting silica beads for the recognition of Human Serum Albumin. Section II: Creation of new imprinted polymers made of polyacrylamide gel capable of removing Human Serum Albumin from human serum, thus increasing the signal of low abundance proteins in two-dimensional maps.