Intramolecular trimerization, a novel strategy for making multispecific antibodies with controlled orientation of the antigen binding domains

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• dc.contributor.author Alvarez-Cienfuegos, Anaca
• dc.contributor.author Nuñez-Prado, Nataliaca
• dc.contributor.author Compte, Martaca
• dc.contributor.author Cuesta, Angel M.ca
• dc.contributor.author Blanco-Toribio, Anaca
• dc.contributor.author Harwood, Seandean Lykkeca
• dc.contributor.author Villate, Maiderca
• dc.contributor.author Merino, Nekaneca
• dc.contributor.author Bonet Martínez, Jaume, 1982-ca
• dc.contributor.author Navarro, Rocioca
• dc.contributor.author Muñoz-Briones, Claraca
• dc.contributor.author Sørensen, Karen Marie Juulca
• dc.contributor.author Mølgaard, Kasperca
• dc.contributor.author Oliva Miguel, Baldomeroca
• dc.contributor.author Sanz, Lauraca
• dc.contributor.author Blanco, Franciscoca
• dc.contributor.author Alvarez-Vallina, Luisca
• dc.date.accessioned 2017-03-10T11:14:11Z
• dc.date.available 2017-03-10T11:14:11Z
• dc.date.issued 2016
• dc.description.abstract Here, we describe a new strategy that allows the rapid and efficient engineering of mono and multispecific trivalent antibodies. By fusing single-domain antibodies from camelid heavy-chain-only immunoglobulins (VHHs) to the N-terminus of a human collagen XVIII trimerization domain (TIEXVIII) we produced monospecific trimerbodies that were efficiently secreted as soluble functional proteins by mammalian cells. The purified VHH-TIEXVIII trimerbodies were trimeric in solution and exhibited excellent antigen binding capacity. Furthermore, by connecting with two additional glycine-serine-based linkers three VHH-TIEXVIII modules on a single polypeptide chain, we present an approach for the rational design of multispecific tandem trimerbodies with defined stoichiometry and controlled orientation. Using this technology we report here the construction and characterization of a tandem VHH-based trimerbody capable of simultaneously binding to three different antigens: carcinoembryonic antigen (CEA), epidermal growth factor receptor (EGFR) and green fluorescence protein (GFP). Multispecific tandem VHH-based trimerbodies were well expressed in mammalian cells, had good biophysical properties and were capable of simultaneously binding their targeted antigens. Importantly, these antibodies were very effective in inhibiting the proliferation of human epidermoid carcinoma A431 cells. Multispecific VHH-based trimerbodies are therefore ideal candidates for future applications in various therapeutic areas.
• dc.description.sponsorship FJB was supported by a grant from the Ministerio de Economía y Competitividad (CTQ2014-56966-R). JB and BO were supported by a grant from the Ministerio de Economía y Competitividad (BIO2011-22568). AA-C and AB-T were supported by Programa Torres Quevedo from Ministerio de Economía y Competitividad, cofounded by the European Social Fund (PTQ09-01-01089 and PTQ11–04604, respectively). SLH is a recipient of the 2016 Novo Scholarship.
• dc.format.mimetype application/pdfca
• dc.identifier.citation Alvarez-Cienfuegos A, Nuñez-Prado N, Compte M, Cuesta AM, Blanco-Toribio A, Harwood SL et al. Intramolecular trimerization, a novel strategy for making multispecific antibodies with controlled orientation of the antigen binding domains. Scientific Reports. 2016;6:28643. DOI: 10.1038/srep28643
• dc.identifier.doi http://dx.doi.org/10.1038/srep28643
• dc.identifier.issn 2045-2322
• dc.identifier.uri http://hdl.handle.net/10230/28206
• dc.language.iso eng
• dc.publisher Nature Publishing Groupca
• dc.relation.ispartof Scientific Reports. 2016;6:28643
• dc.relation.projectID info:eu-repo/grantAgreement/ES/3PN/BIO2011-22568
• dc.rights © Nature Publishing Group. This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/
• dc.rights.accessRights info:eu-repo/semantics/openAccess
• dc.rights.uri http://creativecommons.org/licenses/by/4.0/
• dc.subject.keyword Cancer immunotherapy
• dc.subject.keyword Immunotherapy
• dc.subject.keyword Protein design
• dc.title Intramolecular trimerization, a novel strategy for making multispecific antibodies with controlled orientation of the antigen binding domainsca
• dc.type info:eu-repo/semantics/article
• dc.type.version info:eu-repo/semantics/publishedVersion