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Fast isolation of sub-nanomolar affinity alpaca nanobody against the Spike RBD of SARS-CoV-2 by combining bacterial display and a simple single-step density gradient selection

AuthorsValenzuela Nieto, Guillermo E.; Jara, Ronald; Himelreichs, Johanna; Salinas, Constanza; Pinto, Teresa; Cheuquemilla, Yorka; Margolles, Yago; López González del Rey, Natalia; Chacon, Zaray Miranda; Cuevas, Alexei; Berking, Anne; Deride, Camila; Gonzalez-Moraga, Sebastian; Mancilla, Hector; Maturana, Daniel; Langer, Andreas; Toledo, Juan Pablo; Müller, Ananda; Uberti, Benjamin; Krall, Paola; Ehrenfeld, Pamela; Blesa, Javier; Chana-Cuevas, Pedro; Rehren, German; Fernández, Luis Ángel ; Rojas-Fernandez, Alejandro
Issue Date10-Jun-2020
AbstractDespite the worldwide efforts to avoid disease progression of COVID-19 into a severe acute respiratory syndrome and avoid its severe impact on health systems; the situation remains critical. Effective diagnosis, treatment, and prophylactic measures are required to meet the worldwide demand: recombinant antibodies such as alpaca Nanobodies fulfill these requirements. Here, we develop a fast track for nanobody isolation against the receptor-binding-domain (RBD) SARS-CoV-2 Spike protein following an optimized immunization, efficient construction of the VHH library for E. coli surface display, and single-step selection of high-affinity nanobodies using a simple density gradient centrifugation of the bacterial library. Following this procedure, we isolate and characterize an alpaca Nanobody against Spike RBD of SARS-CoV-2 in the sub-nanomolar range.
Publisher version (URL)https://doi.org/10.1101/2020.06.09.137935
Appears in Collections:(CNB) Artículos
(VICYT) Colección Especial COVID-19
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