Identification of single amino acid differences in uniformly charged homopolymeric peptides with aerolysin nanopore

Fabien Piguet (1) , Hadjer Ouldali (1) , Manuela Pastoriza-Gallego (1) , Philippe Manivet (2, 3) , Juan Pelta (1) , Abdelghani Oukhaled (1)

1 LAMBE - Laboratoire Analyse et Modélisation pour la Biologie et l'Environnement
2 Hôpital Lariboisière-Fernand-Widal [APHP]
3 MASCOT (UMR_S_942 / U942) - Marqueurs cardiovasculaires en situation de stress

There are still unmet needs in finding new technologies for biomedical diagnostic and industrial applications. A technology allowing the analysis of size and sequence of short peptide molecules of only few molecular copies is still challenging. The fast, low-cost and label-free single-molecule nanopore technology could be an alternative for addressing these critical issues. Here, we demonstrate that the wild-type aerolysin nanopore enables the size-discrimination of several short uniformly charged homopeptides, mixed in solution, with a single amino acid resolution. Our system is very sensitive, allowing detecting and characterizing a few dozens of peptide impurities in a high purity commercial peptide sample, while conventional analysis techniques fail to do so.

Mots clés: nanopore; Bacterial Toxins; chemistry; peptide analysis; peptide; aerolysin; bacterial toxin; polymer; pore forming cytotoxic protein; molecular analysis; technology; toxin; analytic method; Article; controlled study; intermethod comparison; particle size; lysine; arginine; amino acid; Nanopores; Nanotechnology; Peptides; Polymers; Pore Forming Cytotoxic Proteins

Domaines: Chimie théorique et/ou physique; Chimie-Physique [physics.chem-ph]

Fabien Piguet, Hadjer Ouldali, Manuela Pastoriza-Gallego, Philippe Manivet, Juan Pelta, et al.. Identification of single amino acid differences in uniformly charged homopolymeric peptides with aerolysin nanopore. Nature Communications, 2018, 9 (1), pp.966. ⟨10.1038/S41467-018-03418-2⟩. ⟨hal-02110440⟩