Yeast surface display platform for rapid discovery of conformationally selective nanobodies

By Conor McMahon, Alexander S. Baier, R. Pascolutti, M. Wegrecki, Sanduo Zheng, Janice X. Ong, S. Erlandson, D. Hilger, S. Rasmussen, A. Ring and 2 more. 2018

467 citations recorded by the source corpus.

Camelid single-domain antibody fragments (‘nanobodies’) provide the remarkable specificity of antibodies within a single 15-kDa immunoglobulin VHH domain. This unique feature has enabled applications ranging from use as biochemical tools to therapeutic agents. Nanobodies have emerged as especially useful tools in protein structural biology, facilitating studies of conformationally dynamic proteins such as G-protein-coupled receptors (GPCRs). Nearly all nanobodies available to date have been obtained by animal immunization, a bottleneck restricting many applications of this technology. To solve this problem, we report a fully in vitro platform for nanobody discovery based on yeast surface display. We provide a blueprint for identifying nanobodies, demonstrate the utility of the library by crystallizing a nanobody with its antigen, and most importantly, we utilize the platform to discover conformationally selective nanobodies to two distinct human GPCRs. To facilitate broad deployment of this platform, the library and associated protocols are freely available for nonprofit research. Yeast surface display platform allows nanobody discovery within two to three weeks. Examples include nanobodies for crystallographic applications, targeting nonpurified antigen or conformationally selective nanobodies to two distinct human GPCRs.

Subfields: Molecular Biology, Radiology, Nuclear Medicine and Imaging

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Citations in the corpus

Accelerating Protein Design Using Autoregressive Generative Models (2019, background): “Recently, phage and yeast display methods have allowed for high-throughput screening of libraries of antibodies and nanobodies (S¨all et al., 2016; McMahon et al., 2018).”
Protein design and variant prediction using autoregressive generative models (2021, methodology): “Using these designed CDR3 sequences, a nanobody library was constructed using our yeast-display technology for experimental characterization alongside a combinatorial synthetic nanobody library 8 .”
Rapid generation of potent antibodies by autonomous hypermutation in yeast (2020, methodology): “Starting from an opensource naïve nanobody YSD library (19), we selected 8 clones that bound the receptor-binding domain (RBD) of the SARS-CoV-2 spike (S) protein (Fig.”