Quantification of Protein Glycosylation Using Nanopores

Roderick Corstiaan Abraham Versloot, Florian Leonardus Rudolfus Lucas, Liubov Yakovlieva, Matthijs Jonathan Tadema, Yurui Zhang, Thomas M Wood, Nathaniel I Martin, Siewert J Marrink, Marthe T C Walvoort*, Giovanni Maglia*

*Corresponding author for this work

Research output: Contribution to journalArticleAcademicpeer-review

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Although nanopores can be used for single-molecule sequencing of nucleic acids using low-cost portable devices, the characterization of proteins and their modifications has yet to be established. Here, we show that hydrophilic or glycosylated peptides translocate too quickly across FraC nanopores to be recognized. However, high ionic strengths (i.e., 3 M LiCl) and low pH (i.e., pH 3) together with using a nanopore with a phenylalanine at its constriction allows the recognition of hydrophilic peptides, and to distinguish between mono- and diglycosylated peptides. Using these conditions, we devise a nanopore method to detect, characterize, and quantify post-translational modifications in generic proteins, which is one of the pressing challenges in proteomic analysis.

Original languageEnglish
Article number2c10338
Pages (from-to)5357-5364
Number of pages8
JournalNano Letters
Issue number13
Early online date29-Jun-2022
Publication statusPublished - 13-Jul-2022


  • protein glycosylation
  • single molecule
  • nanopore spectrometry
  • rhamnosylation
  • proteomics

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