SPR detection of human hepcidin-25: A critical approach by immuno- and biomimetic-based biosensing

• Published in: Biosensors & bioelectronics 2013-02, Vol.40 (1), p.135-140
• Main Authors: Scarano, S., Vestri, A., Ermini, M.L., Minunni, M.
• Format: Article
• Language: English
• Subjects: Anti-hepcidin IgG; Antimicrobial Cationic Peptides - blood; Biological and medical sciences; Biomimetic Materials; Biosensing Techniques - instrumentation; Biosensors; Biotechnology; Equipment Design; Equipment Failure Analysis; Fundamental and applied biological sciences. Psychology; HBD; Hepcidin; Hepcidin binding domain; Hepcidins; Humans; Immunoassay - instrumentation; Iron regulation; Methods. Procedures. Technologies; Reproducibility of Results; Sensitivity and Specificity; SPR; Surface plasmon resonance; Surface Plasmon Resonance - instrumentation; Various methods and equipments
• ISSN: 0956-5663; 1873-4235
• DOI: 10.1016/j.bios.2012.06.060

The human hepcidin-25 hormone has a key role in iron regulation in blood. The clinical relevance of this hepatic ∼2.8kDa cysteine-rich peptide is rapidly increasing, since altered levels can be associated with inflammatory events and iron dysfunctions, such as hereditary hemochromatosis and iron overload. Moreover, hepcidin has also attracted the anti-doping field for its possible role as indirect marker of erythropoietin blood doping. Methods currently reported are based on immunoassays (ELISA and RIA), or various types of mass spectroscopy (MS)-based protocols, semi-quantitative or quantitative. Despite the great effort in optimizing robust and simple assays measuring hepcidin in real matrices, at present this challenge remains still an open issue. To explore the possibility to face hepcidin detection through the development of affinity-based biosensors, we set up a comparative study by surface plasmon resonance (SPR) technology. An immuno-based, on anti-hepcidin-25 IgG, and a biomimetic-based, on a synthetic peptide corresponding to the hepcidin-binding site on ferroportin (HBD), biosensors were developed. Here we report behaviors and analytical performances of the two systems, discussing limits and potentialities. ► SPR applied to human hepcidin-25 detection in standard conditions. ► Hepcidin proper handling was investigated. ► An antibody and a biomimetic receptor were studied and characterized by SPR. ► Experimental detection limits compatible to further application to real samples.