Early diagnosis and efficacious therapy of prostate carcinoma by conventional means still represent daunting challenges. New biotechnology-based reagents and strategies are therefore needed to complement more established forms of tumor management in these areas. Nanoparticles have recently emerged as one of the most promising tools in the oncologist’s armamentarium owing to their advantageous properties as carriers of biologicals and to the ease of detection by highly sensitive analytical devices. In the proposed research we will evaluate the feasibility of in vitro and in vivo targeting of tumor cells by antibody-functionalized nanoparticles with the goal of developing immunodiagnostic and immunotherapeutic tools. Key-points for the success of our investigation will be 1) the use of innovative highly efficient techniques for the obtainment of nanoparticles and for their functionalization with Raman-active reporter molecules and the cross-linking of targeting moieties (monoclonals, recombinant antibody fragments), 2) the availability of high affinity monoclonals and recombinant antibody fragments (scFvs) to a tumor associated biomarker (Prostate Specific Membrane Antigen) and 3) appropriate in vivo mouse models for the assessment of the immunodiagnostic and immunotherapeutic potential of antibody-driven nanoparticles in superficial and metastatic tumors. PSMA was chosen for its wide distribution in prostate tumor (about 70-80% of patients are PSMA +). Anti PSMA radionuclide labelled antibody are under investigation for PCa imaging and therapy. We have therefore investigated the binding properties of gold-NP mAb conjugated by FACS assay; the MFI (mean fluorescence value) measured was of 1.383 for functionalized NP whereas the MFI of the negative control was 68. Selective binding and internalization of mAb-NP were assayed by fluorescence microscopy and by Surface Enhanced Raman Scattering on PSMA positive and negative cells.. We will aim at developing nanoparticles-based immunoimaging protocols and at evaluating therapeutic hyperthermic effects mediated by antibody-driven nanoparticles.
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