A Two-Step Approach for the Design and Generation of Nanobodies.
Fiche publication
Date publication
novembre 2018
Journal
International journal of molecular sciences
Auteurs
Membres identifiés du Cancéropôle Est :
Pr WEISS Etienne
Tous les auteurs :
Wagner HJ, Wehrle S, Weiss E, Cavallari M, Weber W
Lien Pubmed
Résumé
Nanobodies, the smallest possible antibody format, have become of considerable interest for biotechnological and immunotherapeutic applications. They show excellent robustness, are non-immunogenic in humans, and can easily be engineered and produced in prokaryotic hosts. Traditionally, nanobodies are selected from camelid immune libraries involving the maintenance and treatment of animals. Recent advances have involved the generation of nanobodies from naïve or synthetic libraries. However, such approaches demand large library sizes and sophisticated selection procedures. Here, we propose an alternative, two-step approach for the design and generation of nanobodies. In a first step, complementarity-determining regions (CDRs) are grafted from conventional antibody formats onto nanobody frameworks, generating weak antigen binders. In a second step, the weak binders serve as templates to design focused synthetic phage libraries for affinity maturation. We validated this approach by grafting toxin- and hapten-specific CDRs onto frameworks derived from variable domains of camelid heavy-chain-only antibodies (VHH). We then affinity matured the hapten binder via panning of a synthetic phage library. We suggest that this strategy can complement existing immune, naïve, and synthetic library based methods, requiring neither animal experiments, nor large libraries, nor sophisticated selection protocols.
Mots clés
VHH, complementarity-determining region (CDR) grafting, hapten, phage display, single-domain antibody, synthetic library, toxin
Référence
Int J Mol Sci. 2018 Nov 2;19(11):