Solid-phase synthesis of imprinted nanoparticles exhibiting glycan epitope recognition in glycoproteins

Obiettivo

The present project aims at the preparation of synthetic nanoreceptors exhibiting glycan epitope recognition in glycoproteins, identified to play a critical role in cell biology and diseases. Such nanoreceptors, behaving like antibodies, may cope with the current limitations in the essential step of enrichment of specific glycan subclasses, within studies of the glycoproteome. The preparation of the receptors will proceed through solid-phase synthesis of imprinted polymer nanoparticles, with the potential to deliver entities with size, specificity and solubility characteristics comparable to antibodies. The solid-phase synthesis of NanoMIP will require the immobilization of important monosaccharides (N-acetylglucosamine, N-acetlgalactosamine, mannose), widely found in glycoproteins, onto supporting beads. Several synthetic routes will be studied for the immobilization onto silica or plastic beads . In a reactor, built as a controlled temperature glass column filled with the beads containing immobilized monosaccharide, criteriously selected polymerization or sol-gel mixtures will generate the nanoMIPs, with subsequent selection of the highest affinity particles. The optimum combinations of solid-phase/imprinting protocol will be screened from a myriad of nanoMIPs prepared in different conditions. The performance of the nanoMIPs will be ascertained by setting a competitive enrichment protocol consisting of the incubation with a mixture of N-acetylglucosamine-, N-acetylgalactosamine- and mannose-containing glyproteins and a non-glycoprotein. Sorption isotherm data will be used to derive important binding features such as capacity, binding affinity and selectivity. Patentable robust solid-phases and imprinting protocols, either in standalone or combined manner, are planned outputs of the project, which may, in the near future, be incorporated into existing automated solid-phase synthesis instrumentation, for increased throughput and batch reproducibility.