학술논문
Potent neutralization of clinical isolates of SARS-CoV-2 D614 and G614 variants by a monomeric, sub-nanomolar affinity nanobody
Document Type
article
Author
Guillermo Valenzuela Nieto; Ronald Jara; Daniel Watterson; Naphak Modhiran; Alberto A. Amarilla; Johanna Himelreichs; Alexander A. Khromykh; Constanza Salinas-Rebolledo; Teresa Pinto; Yorka Cheuquemilla; Yago Margolles; Natalia López González del Rey; Zaray Miranda-Chacon; Alexei Cuevas; Anne Berking; Camila Deride; Sebastián González-Moraga; Héctor Mancilla; Daniel Maturana; Andreas Langer; Juan Pablo Toledo; Ananda Müller; Benjamín Uberti; Paola Krall; Pamela Ehrenfeld; Javier Blesa; Pedro Chana-Cuevas; German Rehren; David Schwefel; Luis Ángel Fernandez; Alejandro Rojas-Fernandez
Source
Scientific Reports, Vol 11, Iss 1, Pp 1-14 (2021)
Subject
Language
English
ISSN
2045-2322
Abstract
Abstract 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.