title: Screening of the binding properties of molecularly imprinted nanoparticles via capillary electrophoresis.
abstract: In response to the need for straightforward analytical methods to assess the affinity of molecularly imprinted nanoparticles (MIP NPs) for ligands, capillary electrophoresis (CE) was exploited using MIP NPs targeting the iron-regulating hormone hepcidin. In this work, MIP NPs were challenged with their template peptide, i.e., the N-terminal 5-mer of hepcidin, in comparison to unrelated ligand peptides. A CE separation method was developed ex novo achieving, after optimization of the background electrolyte (150 mM sodium phosphate pH 7.4) and of the running temperature (35 °C), the full separation of the free ligand from the complexed MIP NPs. The CE binding isotherm allowed the estimation of a micromolar dissociation constant for the 5-mer template-MIP NPs complex, in agreement with independent measurements. The CE offered the advantages of a direct injection of the MIP NPs/ligand incubation mix, without preliminary fractionation steps, requiring only minimal sample volumes and short analysis times. In conclusion CE proved to be a valid technique for characterizing the interactions of MIP NP libraries for selected target compounds.
authors.affiliation: Department of Diagnostics and Public Health, Unit of Forensic Medicine, University of Verona, Piazzale Ludovico Antonio Scuro 10, 37134, Verona, Italy.
authors.forename: Giacomo
authors.initials: G
authors.lastname: Musile
authors.affiliation: Department of Biotechnology, University of Verona, Strada Le Grazie 15, 37134, Verona, Italy.
authors.forename: Lucia
authors.initials: L
authors.lastname: Cenci
authors.affiliation: Department of Biotechnology, University of Verona, Strada Le Grazie 15, 37134, Verona, Italy.
authors.forename: Erika
authors.initials: E
authors.lastname: Andreetto
authors.affiliation: Department of Molecular Sciences and Nanosystems, University Ca' Foscari Venezia, via Torino 155/b, 30173, Venice, Italy.
authors.forename: Emmanuele
authors.initials: E
authors.lastname: Ambrosi
authors.affiliation: Department of Diagnostics and Public Health, Unit of Forensic Medicine, University of Verona, Piazzale Ludovico Antonio Scuro 10, 37134, Verona, Italy.
authors.forename: Franco
authors.initials: F
authors.lastname: Tagliaro
authors.affiliation: Department of Biotechnology, University of Verona, Strada Le Grazie 15, 37134, Verona, Italy. alessandramaria.bossi@univr.it.
authors.forename: Alessandra Maria
authors.initials: AM
authors.lastname: Bossi
mesh_terms: D001665:Binding Sites; D002851:Chromatography, High Pressure Liquid; D019075:Electrophoresis, Capillary / Q000379:methods*; D006863:Hydrogen-Ion Concentration; D008024:Ligands; D054802:Molecular Imprinting / Q000379:methods*; D053758:Nanoparticles; D013696:Temperature
chemical_list: D008024:Ligands
keywords: Affinity; Binding isotherm; Capillary electrophoresis; Hepcidin; Molecularly imprinted nanoparticles