Journal
JOURNAL OF NANOBIOTECHNOLOGY
Volume 13, Issue -, Pages -Publisher
BMC
DOI: 10.1186/s12951-015-0115-3
Keywords
Molecularly imprinted polymers; Nanoparticles; Hepcidin; Biosensor; Surface plasmon resonance; Iron metabolism
Funding
- Italian Ministry of Education, Research and University [FIRB2012 RBFR12LD0W_003, 200989KXFN_002]
- Fondazione Cariverona
- People Programme (Marie Curie Actions, PINP project) of the European Union's Seventh Framework Programme under REA grant [327165]
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Background: Molecularly imprinted polymer (MIP) technique is a powerful mean to produce tailor made synthetic recognition sites. Here precipitation polymerization was exploited to produce a library of MIP nanoparticles (NPs) targeting the N terminus of the hormone Hepcidin-25, whose serum levels correlate with iron dis-metabolisms and doping. Biotinylated MIP NPs were immobilized to NeutrAvidin (TM) SPR sensor chip. The response of the MIP NP sensor to Hepcidin-25 was studied. Findings: Morphological analysis showed MIP NPs of 20-50 nm; MIP NP exhibited high affinity and selectivity for the target analyte: low nanomolar Kds for the interaction NP/Hepcidin-25, but none for the NP/non regulative Hepcidin-20. The MIP NP were integrated as recognition element in SPR allowing the detection of Hepcidin-25 in 3 min. Linearity was observed with the logarithm of Hepcidin-25 concentration in the range 7.2-720 pM. LOD was 5 pM. The response for Hepcidin-20 was limited. Hepcidin-25 determination in real serum samples spiked with known analyte concentrations was also attempted. Conclusion: The integration of MIP NP to SPR allowed the determination of Hepcidin-25 at picomolar concentrations in short times outperforming the actual state of art. Optimization is still needed for real sample measurements in view of future clinical applications.
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