Selective protein detection with a dsLNA-functionalized nanopore

In the last years, nanopore technology has been increasingly exploited for biomolecule detection andanalysis. Recently, the main focus of the research has moved from the study of nucleic acids to theanalysis of proteins and DNA-protein complexes. In this paper, chemically functionalized solid-statenanopore has been used to recognize Nuclear Factor-kappa B proteins (NF-κB), that are involved inseveral disorders and inflammation processes, so that their identification is of crucial importance forprognostic applications. In particular, we show that it is possible to electrically detect the specificinteraction between p50, a protein belonging to the NF-κB family, and dsLNA probe molecules covalentlyattached to the surface of a FIB fabricated SiN pore. The obtained results have been compared with thoserelated to BSA protein, which does not interact with the used probes. Finally, the potential of the devicehas been further tested by analyzing a whole cell extract. In this case, three principal peaks in thedistribution of electrical event duration can be identified, corresponding to different interacting NF-κBcomplexes, so that the methodology appears to be effective also to study biological samples ofconsiderable complexity. Ultimately, the presented data emphasize the selectivity and versatility ofthe functionalized nanopore device, demonstrating its applicability in bioanalytics and advanceddiagnostics.