open access publication

Article, 2023

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

In: Chemistry - An Asian Journal, ISSN 1861-4728, 1861-471X, Page e202300515, 10.1002/asia.202300515

Contributors (5)

Salehirozveh, Mostafa (0000-0002-2104-7735) [1] [2] Larsen, Anne-Kathrine Kure [3] [4] [5] Stojmenovic, Milos [6] Thei, Federico (0000-0002-3959-2002) (Corresponding author) [2] Dong, Ming-Dong (0000-0002-2025-2171) (Corresponding author) [5]


  1. [1] University of Bologna
  2. [NORA names: Italy; Europe, EU; OECD]
  3. [2] Elements srl, Cesena, Italy
  4. [3] Sino-Danish Center for Education and Research
  5. [NORA names: Miscellaneous]
  6. [4] University of Chinese Academy of Sciences
  7. [NORA names: China; Asia, East]
  8. [5] Aarhus University
  9. [NORA names: AU Aarhus University; University; Denmark; Europe, EU; Nordic; OECD]


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.


AAT protein, Functionalized Nanopores, Herein, PEG approach, PLL, PMMA, addition, adequate functionalization, alpha-1 antitrypsin (AAT) protein, amplitude, antitrypsin protein, applications, approach, biomarkers, characteristics, characterization, continuous monitoring, detection, detection technology, disease-relevant biomarkers, dwell time, events, experiments, flowcell, functionalization, functionalization method, great promise, hydrodynamic volume, increase, individual proteins, label-free detection, lifetime, method, monitoring, nanopore applications, nanopore experiments, nanopores, number, promise, protein, protein characterization, protein volume, proteomics research, reduction, research, single-molecule characterization, situ, situ functionalization method, stimuli, study, technology, time, translocation events, translocation experiments, voltage stimuli, volume


  • Danish Agency for Science and Higher Education
  • European Commission