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
| Autor: | Valenzuela 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 |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2020 |
| Zdroj: | Digital.CSIC. Repositorio Institucional del CSIC instname |
| Popis: | Despite 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. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|