title: Selective protein detection with a dsLNA-functionalized nanopore.
abstract: In the last years, nanopore technology has been increasingly exploited for biomolecule detection and analysis. Recently, the main focus of the research has moved from the study of nucleic acids to the analysis of proteins and DNA-protein complexes. In this paper, chemically functionalized solid-state nanopore has been used to recognize Nuclear Factor-kappa B proteins (NF-κB), that are involved in several disorders and inflammation processes, so that their identification is of crucial importance for prognostic applications. In particular, we show that it is possible to electrically detect the specific interaction between p50, a protein belonging to the NF-κB family, and dsLNA probe molecules covalently attached to the surface of a FIB fabricated SiN pore. The obtained results have been compared with those related to BSA protein, which does not interact with the used probes. Finally, the potential of the device has been further tested by analyzing a whole cell extract. In this case, three principal peaks in the distribution of electrical event duration can be identified, corresponding to different interacting NF-κB complexes, so that the methodology appears to be effective also to study biological samples of considerable complexity. Ultimately, the presented data emphasize the selectivity and versatility of the functionalized nanopore device, demonstrating its applicability in bioanalytics and advanced diagnostics.
authors.affiliation: Department of Physics, University of Genoa, via Dodecaneso 33, 16146 Genova, Italy.
authors.forename: Paola
authors.initials: P
authors.lastname: Fanzio
authors.affiliation: National Research Council, Institute for Complex Systems ISC-CNR, Via del Fosso del Cavaliere 100, 00133 Roma, Italy.
authors.forename: Valentina
authors.initials: V
authors.lastname: Mussi
authors.affiliation: Department of Biomolecular Sciences, University of Urbino 'Carlo Bo', Via Saffi 2, 61029 Urbino, PU, Italy.
authors.forename: Michele
authors.initials: M
authors.lastname: Menotta
authors.affiliation: Department of Physics, University of Genoa, via Dodecaneso 33, 16146 Genova, Italy.
authors.forename: Giuseppe
authors.initials: G
authors.lastname: Firpo
authors.affiliation: Department of Physics, University of Genoa, via Dodecaneso 33, 16146 Genova, Italy.
authors.forename: Luca
authors.initials: L
authors.lastname: Repetto
authors.affiliation: Department of Physics, University of Genoa, via Dodecaneso 33, 16146 Genova, Italy.
authors.forename: Patrizia
authors.initials: P
authors.lastname: Guida
authors.affiliation: Department of Physics, University of Genoa, via Dodecaneso 33, 16146 Genova, Italy.
authors.forename: Elena
authors.initials: E
authors.lastname: Angeli
authors.affiliation: Department of Biomolecular Sciences, University of Urbino 'Carlo Bo', Via Saffi 2, 61029 Urbino, PU, Italy.
authors.forename: Mauro
authors.initials: M
authors.lastname: Magnani
authors.affiliation: Department of Physics, University of Genoa, via Dodecaneso 33, 16146 Genova, Italy.
authors.forename: Ugo
authors.initials: U
authors.lastname: Valbusa
mesh_terms: D015374:Biosensing Techniques / Q000295:instrumentation*; D003217:Conductometry / Q000295:instrumentation*; D004867:Equipment Design; D019544:Equipment Failure Analysis; D016328:NF-kappa B / Q000032:analysis* / Q000737:chemistry / Q000235:genetics; D058608:Nanopores / Q000648:ultrastructure*; D009841:Oligonucleotides / Q000737:chemistry* / Q000235:genetics; D040081:Protein Array Analysis / Q000295:instrumentation*
chemical_list: D016328:NF-kappa B; D009841:Oligonucleotides; C477371:locked nucleic acid
keywords: Biosensor; Functionalization; NF-κB; Nanopore; Protein detection