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Abstract
The grafting reaction of oligodeoxyribonucleotides (ODN) on N-vinyl pyrrolidone (NVP)/N-acryloxy succinimide (NAS) copolymers has been investigated. First, NVP/NAS copolymers were prepared by free-radical polymerization in N,N-dimethylformamide (DMF); characterization of the monomer sequence distribution by 13C NMR suggested a strongly alternating structure. Low-conversion copolymers were synthesized using azeotropic composition (60% molar NAS) and various overall monomer concentrations. These model copolymers were characterized by light scattering and viscosity measurements in a DMF-water (90/10 volume ratio) mixture.
The covalent attachment of ODN (constituted of 20 bases terminated with a primary amine linker) onto one selected copolymer (60% molar NAS) was then examined. Size exclusion chromatography (SEC) and free solution capillary electrophoresis (FSCE) were performed in order to analyse the reaction products and to calculate the yield of grafting. After optimization of the immobilization conditions, the influence of buffer pH and temperature on the grafting reaction kinetics was considered.