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Synthesis of peptide imprinted nanoparticles and their integration to ELISA-like assay for the quantification of hepcidin (PINP)
Start date: May 1, 2013, End date: Apr 30, 2015 PROJECT  FINISHED 

"The development of assays based on molecular recognition for measuring biomarkers is a critical task for the diagnosis and prognosis of pathologies. The present project aims at developing synthetic nanoparticles (NP) for the quantification of the peptide biomarker hepcidin (hep) which regulates iron homeostasis. A modern strategies for mimicking receptors consists of NPs produced by molecularly imprinted polymer (MIP) technology. MIPs are prepared by a template-assisted emulsion polymerization. After removal of the template, complementary cavities are exposed in the polymer. The target biomarker, hep, is a 25-residue peptide (hep25) present in urine and plasma, other N-terminus truncated isoforms such as hep20 and hep22 were also found in humans. It is known that the first 5 amino acids of the N-terminus of hep25 are essential for iron homeostasis while the role of other isoforms is still unclear. Given that the imprinting is more successful for short peptides only 2 crucial portions of the full length peptide are synthesized as templates which are also functionalized to impose a direction of imprinting. The precision of the template synthesis ensures a higher fidelity of imprinting during the NPs synthesis improving the molecular recognition. The goal of the project consist in designing an assay for the determination of serum and/or urinary hep, based on MIPs selective for hep isoforms and arranged in a pseudo-immuno assay format, further extended in sensitivity by the addition of fluorescent tags. At last, an evaluation of the assay with clinical sample is planned to be performed. The possibility of using nano-sized MIPs as plastic antibodies in ELISA-like assays has the obvious advantage of a potential widespread diffusion. The proposed ELISA-like assay with fluorescent tags is very innovative and have the potential versatility to be transposed to different applications. Moreover the deep impact of the outcomes allow the access to the diagnostic market."

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