Potent neutralization of clinical isolates of SARS-CoV-2 D614 and G614 variants by a monomeric, sub-nanomolar affinity nanobody.

Autor: Valenzuela Nieto G; Institute of Medicine, Faculty of Medicine, Universidad Austral de Chile, Valdivia, Chile., Jara R; Institute of Medicine, Faculty of Medicine, Universidad Austral de Chile, Valdivia, Chile.; Institute of Biochemistry and Microbiology, Faculty of Sciences, Universidad Austral de Chile, Valdivia, Chile., Watterson D; School of Chemistry and Molecular Bioscience, The University of Queensland, Brisbane, Australia.; The Australian Institute for Biotechnology and Nanotechnology, The University of Queensland, Brisbane, Australia.; Australian Infectious Diseases Research Centre, The University of Queensland, Brisbane, Australia., Modhiran N; School of Chemistry and Molecular Bioscience, The University of Queensland, Brisbane, Australia.; The Australian Institute for Biotechnology and Nanotechnology, The University of Queensland, Brisbane, Australia., Amarilla AA; School of Chemistry and Molecular Bioscience, The University of Queensland, Brisbane, Australia., Himelreichs J; Institute of Medicine, Faculty of Medicine, Universidad Austral de Chile, Valdivia, Chile., Khromykh AA; School of Chemistry and Molecular Bioscience, The University of Queensland, Brisbane, Australia.; Australian Infectious Diseases Research Centre, The University of Queensland, Brisbane, Australia., Salinas-Rebolledo C; Institute of Medicine, Faculty of Medicine, Universidad Austral de Chile, Valdivia, Chile., Pinto T; Institute of Medicine, Faculty of Medicine, Universidad Austral de Chile, Valdivia, Chile., Cheuquemilla Y; Institute of Medicine, Faculty of Medicine, Universidad Austral de Chile, Valdivia, Chile.; Berking Biotechnology, Valdivia, Chile., Margolles Y; Department of Microbial Biotechnology, National Biotechnology Center, Superior Council of Scientific Research, Madrid, Spain., López González Del Rey N; HM CINAC, Hospital Universitario HM Puerta del Sur, Mostoles, 28938, Madrid, Spain., Miranda-Chacon Z; Institute of Medicine, Faculty of Medicine, Universidad Austral de Chile, Valdivia, Chile., Cuevas A; Institute of Medicine, Faculty of Medicine, Universidad Austral de Chile, Valdivia, Chile., Berking A; Berking Biotechnology, Valdivia, Chile., Deride C; Institute of Medicine, Faculty of Medicine, Universidad Austral de Chile, Valdivia, Chile.; Institute of Veterinary Clinical Sciences, Faculty of Veterinary Sciences, Universidad Austral de Chile, Valdivia, Chile., González-Moraga S; Institute of Medicine, Faculty of Medicine, Universidad Austral de Chile, Valdivia, Chile., Mancilla H; Institute of Medicine, Faculty of Medicine, Universidad Austral de Chile, Valdivia, Chile., Maturana D; NanoTemper Technologies GmbH, Floessergasse 4, 81369, Munich, Germany., Langer A; NanoTemper Technologies GmbH, Floessergasse 4, 81369, Munich, Germany., Toledo JP; Institute of Medicine, Faculty of Medicine, Universidad Austral de Chile, Valdivia, Chile., Müller A; Ross University School of Veterinary Medicine, Basseterre, Saint Kitts and Nevis.; Institute of Veterinary Clinical Sciences, Faculty of Veterinary Sciences, Universidad Austral de Chile, Valdivia, Chile., Uberti B; Institute of Veterinary Clinical Sciences, Faculty of Veterinary Sciences, Universidad Austral de Chile, Valdivia, Chile., Krall P; Institute of Medicine, Faculty of Medicine, Universidad Austral de Chile, Valdivia, Chile.; Department of Pediatrics and Children's Surgery Oriente, Universidad de Chile, Valdivia, Chile., Ehrenfeld P; Institute of Anatomy, Histology, and Pathology, Faculty of Medicine, Universidad Austral de Chile, Valdivia, Chile.; Center for Interdisciplinary Studies on the Nervous System, CISNE, Universidad Austral de Chile, Valdivia, Chile., Blesa J; HM CINAC, Hospital Universitario HM Puerta del Sur, Mostoles, 28938, Madrid, Spain., Chana-Cuevas P; CETRAM & Faculty of Medical Science, Universidad de Santiago de Chile, Santiago, Chile., Rehren G; Technology Transfer and Licensing Office, Universidad Austral de Chile, Valdivia, Chile., Schwefel D; Charité-Universitätsmedizin Berlin, Corporate Member of Freie Universität Berlin, Humboldt-Universität zu Berlin, Berlin Institute of Health, Berlin, Germany., Fernandez LÁ; Department of Microbial Biotechnology, National Biotechnology Center, Superior Council of Scientific Research, Madrid, Spain., Rojas-Fernandez A; Institute of Medicine, Faculty of Medicine, Universidad Austral de Chile, Valdivia, Chile. alejandro.rojas@uach.cl.; Berking Biotechnology, Valdivia, Chile. alejandro.rojas@uach.cl.; Center for Interdisciplinary Studies on the Nervous System, CISNE, Universidad Austral de Chile, Valdivia, Chile. alejandro.rojas@uach.cl.; Institute of Philosophy and Complexity Sciences, Santiago, Chile. alejandro.rojas@uach.cl.
Jazyk: angličtina
Zdroj: Scientific reports [Sci Rep] 2021 Feb 08; Vol. 11 (1), pp. 3318. Date of Electronic Publication: 2021 Feb 08.
DOI: 10.1038/s41598-021-82833-w
Abstrakt: Despite unprecedented global efforts to rapidly develop SARS-CoV-2 treatments, in order to reduce the burden placed on health systems, the situation remains critical. Effective diagnosis, treatment, and prophylactic measures are urgently required to meet global demand: recombinant antibodies fulfill these requirements and have marked clinical potential. Here, we describe the fast-tracked development of an alpaca Nanobody specific for the receptor-binding-domain (RBD) of the SARS-CoV-2 Spike protein with potential therapeutic applicability. We present a rapid method for nanobody isolation that includes an optimized immunization regimen coupled with VHH library E. coli surface display, which allows single-step selection of Nanobodies using a simple density gradient centrifugation of the bacterial library. The selected single and monomeric Nanobody, W25, binds to the SARS-CoV-2 S RBD with sub-nanomolar affinity and efficiently competes with ACE-2 receptor binding. Furthermore, W25 potently neutralizes SARS-CoV-2 wild type and the D614G variant with IC50 values in the nanomolar range, demonstrating its potential as antiviral agent.
Databáze: MEDLINE
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