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Platform for rapid nanobody discovery in vitro

By Conor McMahon, Alexander S Baier, Sanduo Zheng, Roberta Pascolutti, Janice X. Ong, Sarah C. Erlandson, Daniel Hilger, Aaron M. Ring, Aashish Manglik, Andrew C. Kruse

Posted 16 Jun 2017
bioRxiv DOI: 10.1101/151043 (published DOI: 10.1101/151043)

Camelid single-domain antibody fragments ('nanobodies') provide the remarkable specificity of antibodies within a single immunoglobulin VHH domain. This unique feature enables applications ranging from their use as biochemical tools to therapeutic agents. Virtually all nanobodies reported to date have been obtained by animal immunization, a bottleneck restricting many applications of this technology. To solve this problem, we developed a fully in vitro platform for nanobody discovery based on yeast surface display of a synthetic nanobody scaffold. This platform provides a facile and cost-effective method for rapidly isolating nanobodies targeting a diverse range of antigens. We provide a blueprint for identifying nanobodies starting from both purified and non-purified antigens, and in addition, we demonstrate application of the platform to discover rare conformationally-selective nanobodies to a lipid flippase and a G protein-coupled receptor. To facilitate broad deployment of this platform, we have made the library and all associated protocols publicly available.

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