In-situ PLL-g-PEG Functionalized Nanopore for Enhancing Protein Characterization

Mostafa Salehirozveh, Anne Kathrine Kure Larsen, Milos Stojmenovic, Federico Thei*, Mingdong Dong*

*Corresponding author for this work

Research output: Contribution to journal/Conference contribution in journal/Contribution to newspaperJournal articleResearchpeer-review

Abstract

Single-molecule nanopore detection technology has revolutionized proteomics research by enabling highly sensitive and label-free detection of individual proteins. Herein, we designed a small, portable, and leak-free flowcell made of PMMA for nanopore experiments. In addition, we developed an in situ functionalizing PLL-g-PEG approach to produce non-sticky nanopores for measuring the volume of diseases-relevant biomarker, such as the Alpha-1 antitrypsin (AAT) protein. The in situ functionalization method allows continuous monitoring, ensuring adequate functionalization, which can be directly used for translocation experiments. The functionalized nanopores exhibit improved characteristics, including an increased nanopore lifetime and enhanced translocation events of the AAT proteins. Furthermore, we demonstrated the reduction in the translocation event's dwell time, along with an increase in current blockade amplitudes and translocation numbers under different voltage stimuli. The study also successfully measures the single AAT protein volume (253 nm3), which closely aligns with the previously reported hydrodynamic volume. The real-time in situ PLL-g-PEG functionalizing method and the developed nanopore flowcell hold great promise for various nanopores applications involving non-sticky single-molecule characterization.

Original languageEnglish
Article numbere202300515
JournalChemistry - An Asian Journal
Volume18
Issue17
Number of pages11
ISSN1861-4728
DOIs
Publication statusPublished - Sept 2023

Keywords

  • Flowcell
  • In situ functionalization
  • Nanopore
  • Non-sticky
  • Protein

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