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Structure-based design and construction of a synthetic phage display nanobody library

dc.contributor.authorMoreno E
dc.contributor.authorValdés-Tresanco M.S
dc.contributor.authorMolina-Zapata A
dc.contributor.authorSánchez-Ramos O.
dc.date.accessioned2022-09-14T14:34:10Z
dc.date.available2022-09-14T14:34:10Z
dc.date.created2022
dc.identifier.issn17560500
dc.identifier.urihttp://hdl.handle.net/11407/7585
dc.descriptionObjective: To design and construct a new synthetic nanobody library using a structure-based approach that seeks to maintain high protein stability and increase the number of functional variants within the combinatorial space of mutations. Results: Synthetic nanobody (Nb) libraries are emerging as an attractive alternative to animal immunization for the selection of stable, high affinity Nbs. Two key features define a synthetic Nb library: framework selection and CDR design. We selected the universal VHH framework from the cAbBCII10 Nb. CDR1 and CDR2 were designed with the same fixed length as in cAbBCII10, while for CDR3 we chose a 14-long loop, which creates a convex binding site topology. Based on the analysis of the cAbBCII10 crystal structure, we carefully selected the positions to be randomized and tailored the codon usage at each position, keeping at particular places amino acids that guarantee stability, favoring properties like polarity at solvent-exposed positions and avoiding destabilizing amino acids. Gene synthesis and library construction were carried out by GenScript, using our own phagemid vector. The constructed library has an estimated size of 1.75 × 108. NGS showed that the amino acid diversity and frequency at each randomized position are the expected from the codon usage. © 2022, The Author(s).eng
dc.language.isoeng
dc.publisherBioMed Central Ltd
dc.relation.isversionofhttps://www.scopus.com/inward/record.uri?eid=2-s2.0-85127254338&doi=10.1186%2fs13104-022-06001-7&partnerID=40&md5=f1b6a817144ea073bb91b3bd7251c847
dc.sourceBMC Research Notes
dc.titleStructure-based design and construction of a synthetic phage display nanobody library
dc.typeNote
dc.typeother
dc.rights.accessrightsinfo:eu-repo/semantics/restrictedAccess
dc.publisher.programCiencias Básicas
dc.type.spaOtro
dc.identifier.doi10.1186/s13104-022-06001-7
dc.subject.keywordNanobodyeng
dc.subject.keywordPhage displayeng
dc.subject.keywordStructure-based designeng
dc.subject.keywordSynthetic nanobody libraryeng
dc.relation.citationvolume15
dc.relation.citationissue1
dc.publisher.facultyFacultad de Ciencias Básicas
dc.affiliationMoreno, E., Faculty of Basic Sciences, University of Medellin, Medellín, Colombia
dc.affiliationValdés-Tresanco, M.S., Faculty of Basic Sciences, University of Medellin, Medellín, Colombia
dc.affiliationMolina-Zapata, A., Faculty of Basic Sciences, University of Medellin, Medellín, Colombia, Grupo de Micología Médica y Experimental, Corporación para Investigaciones Biológicas (CIB), Medellín, Colombia
dc.affiliationSánchez-Ramos, O., Biological Sciences School, University of Concepción, Concepción, Chile
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dc.type.coarhttp://purl.org/coar/resource_type/c_1843
dc.type.versioninfo:eu-repo/semantics/publishedVersion
dc.type.driverinfo:eu-repo/semantics/other
dc.identifier.reponamereponame:Repositorio Institucional Universidad de Medellín
dc.identifier.repourlrepourl:https://repository.udem.edu.co/
dc.identifier.instnameinstname:Universidad de Medellín


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