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    Rapid single-molecule detection of COVID-19 and MERS antigens via nanobody-functionalized organic electrochemical transistors

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    Accepted Manuscript
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    Type
    Article
    Authors
    Guo, Keying cc
    Wustoni, Shofarul cc
    Koklu, Anil
    Díaz-Galicia, Escarlet cc
    Moser, Maximilian cc
    Hama, Adel
    Alqahtani, Ahmed A.
    Ahmad, Adeel Nazir cc
    Alhamlan, Fatimah Saeed
    Shuaib, Muhammad cc
    Pain, Arnab cc
    McCulloch, Iain cc
    Arold, Stefan T. cc
    Grunberg, Raik cc
    Inal, Sahika cc
    KAUST Department
    Biological and Environmental Science and Engineering (BESE) Division
    Bioscience Program
    Chemical Science Program
    Computational Bioscience Research Center (CBRC)
    KAUST Health, Thuwal, Saudi Arabia.
    KAUST Solar Center (KSC)
    Pathogen Genomics Laboratory
    Physical Science and Engineering (PSE) Division
    Structural Biology and Engineering
    KAUST Grant Number
    OSR-2015-CRG4-2572
    OSR-2018-CARF/CCF-3079
    OSR-2018-CRG7-3709
    OSR-4106 CPF2019
    REI/1/4204-01
    REI/1/4229-01
    Date
    2021-05-24
    Online Publication Date
    2021-05-24
    Print Publication Date
    2021-07
    Embargo End Date
    2021-11-24
    Submitted Date
    2020-11-10
    Permanent link to this record
    http://hdl.handle.net/10754/669265
    
    Metadata
    Show full item record
    Abstract
    The coronavirus disease 2019 (COVID-19) pandemic has highlighted the need for rapid and sensitive protein detection and quantification in simple and robust formats for widespread point-of-care applications. Here, we report on nanobody-functionalized organic electrochemical transistors with a modular architecture for the rapid quantification of single-molecule-to-nanomolar levels of specific antigens in complex bodily fluids. The sensors combine a solution-processable conjugated polymer in the transistor channel and high-density and orientation-controlled bioconjugation of nanobody–SpyCatcher fusion proteins on disposable gate electrodes. The devices provide results after 10 min of exposure to 5 μl of unprocessed samples, maintain high specificity and single-molecule sensitivity in human saliva and serum, and can be reprogrammed to detect any protein antigen if a corresponding specific nanobody is available. We used the sensors to detect green fluorescent protein, and severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) and Middle East respiratory syndrome coronavirus (MERS-CoV) spike proteins, and for the COVID-19 screening of unprocessed clinical nasopharyngeal swab and saliva samples with a wide range of viral loads.
    Citation
    Guo, K., Wustoni, S., Koklu, A., Díaz-Galicia, E., Moser, M., Hama, A., … Inal, S. (2021). Rapid single-molecule detection of COVID-19 and MERS antigens via nanobody-functionalized organic electrochemical transistors. Nature Biomedical Engineering. doi:10.1038/s41551-021-00734-9
    Sponsors
    Figure 1 was produced by X. Pita, a scientific illustrator at KAUST. We thank all of the members of the KAUST Rapid Research Response Team (R3T) for COVID-19, especially S. Hamdan, for contributions in this study. We thank S. Mfarrej and A. K. Subudhi for providing access to and assisting with the experiments in the Biosafety Level 2+ experimental room at KAUST. We thank the KAUST Health team (operated by Dr. Soliman Fakeeh Hospital, Jeddah), including D. Buttigieg and M. Habib, for providing clinical samples. We thank staff at the King Faisal Specialist Hospital and Research Center (Riyadh), particularly A. Alzahrani, M. Alsanea and F. Alhadeq, for help with organizing and hosting some of the clinical studies. We thank the KAUST nanofabrication core laboratory team, D. Rosas Villalva and U. Buttner for help with device fabrication and integration. This work was initiated thanks to the KAUST Impact Acceleration Fund (IAF) program. The research reported in this publication was supported by funding from the Office of Sponsored Research (OSR) at KAUST under award numbers REI/1/4204-01, REI/1/4229-01, OSR-2018-CRG7-3709, OSR-2018-CARF/CCF-3079, OSR-2015-CRG4-2572 and OSR-4106 CPF2019. We acknowledge EC FP7 Project SC2 (610115), EC H2020 (643791) and EPSRC Projects EP/G037515/1, EP/M005143/1 and EP/L016702/1.
    Publisher
    Springer Science and Business Media LLC
    Journal
    Nature Biomedical Engineering
    DOI
    10.1038/s41551-021-00734-9
    10.1101/2020.11.12.20228874
    Additional Links
    http://www.nature.com/articles/s41551-021-00734-9
    ae974a485f413a2113503eed53cd6c53
    10.1038/s41551-021-00734-9
    Scopus Count
    Collections
    Articles; Biological and Environmental Science and Engineering (BESE) Division; Bioscience Program; Physical Science and Engineering (PSE) Division; Chemical Science Program; Computational Bioscience Research Center (CBRC); KAUST Solar Center (KSC)

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