Yeast surface display platform for rapid discovery of conformationally selective nanobodies

TY - JOUR

T1 - Yeast surface display platform for rapid discovery of conformationally selective nanobodies

AU - McMahon, Conor

AU - Baier, Alexander S.

AU - Pascolutti, Roberta

AU - Wegrecki, Marcin

AU - Zheng, Sanduo

AU - Ong, Janice X.

AU - Erlandson, Sarah C.

AU - Hilger, Daniel

AU - Rasmussen, Søren G.F.

AU - Ring, Aaron M.

AU - Manglik, Aashish

AU - Kruse, Andrew C.

PY - 2018

Y1 - 2018

N2 - 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.

AB - 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.

U2 - 10.1038/s41594-018-0028-6

DO - 10.1038/s41594-018-0028-6

M3 - Journal article

C2 - 29434346

AN - SCOPUS:85042625183

SN - 1545-9993

VL - 25

SP - 289

EP - 296

JO - Nature Structural and Molecular Biology

JF - Nature Structural and Molecular Biology

IS - 3

ER -