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Amyloid Growth, Inhibition, and Real-Time Enzymatic Degradation Revealed with Single Conical Nanopore

Author:
Giamblanco, Nicoletta, Coglitore, Diego, Gubbiotti, Alberto, Ma, Tianji, Balanzat, Emmanuel, Janot, Jean-Marc, Chinappi, Mauro, Balme, Sebastien
Source:
Analytical chemistry 2018 v.90 no.21 pp. 12900-12908
ISSN:
1520-6882
Subject:
amyloid, enzymes, fouling, nanopores, neurodegenerative diseases
Abstract:
Amyloid fibrils are involved in several neurodegenerative diseases. However, because of their polymorphism and low concentration, they are challenging to assess in real-time with conventional techniques. Here, we present a new approach for the characterization of the intermediates: protofibrils and “end-off” aggregates which are produced during the amyloid formation. To do so, we have fashioned conical track-etched nanopores that are functionalized to prevent the fouling. Using these nanopores, we have followed the kinetic of amyloid growth to discriminate the different intermediates protofibrils and “end-off. Then, the nanopore was used to characterize the effect of promoter and inhibitor of the fibrillation process. Finally, we have followed in real-time the degradation of amyloid with peptase. Compare with the SiN nanopore, the track-etched one features exceptionally high success rate via functionalization and detection in “one-pot”. Our results demonstrate the potential for a conical nanopore to be used as a routine technique for the characterization of the amyloid growth and/or degradation.
Agid:
6201606