TY - JOUR
T1 - In-situ PLL-g-PEG Functionalized Nanopore for Enhancing Protein Characterization
AU - Salehirozveh, Mostafa
AU - Kure Larsen, Anne Kathrine
AU - Stojmenovic, Milos
AU - Thei, Federico
AU - Dong, Mingdong
N1 - Publisher Copyright:
© 2023 The Authors. Chemistry - An Asian Journal published by Wiley-VCH GmbH.
PY - 2023/9
Y1 - 2023/9
N2 - 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.
AB - 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.
KW - Flowcell
KW - In situ functionalization
KW - Nanopore
KW - Non-sticky
KW - Protein
UR - http://www.scopus.com/inward/record.url?scp=85169514828&partnerID=8YFLogxK
U2 - 10.1002/asia.202300515
DO - 10.1002/asia.202300515
M3 - Journal article
C2 - 37497831
AN - SCOPUS:85169514828
SN - 1861-4728
VL - 18
JO - Chemistry - An Asian Journal
JF - Chemistry - An Asian Journal
IS - 17
M1 - e202300515
ER -