Yeast surface display platform for rapid discovery of conformationally selective nanobodies
Research output: Contribution to journal › Journal article › Research › peer-review
Standard
Yeast surface display platform for rapid discovery of conformationally selective nanobodies. / McMahon, Conor; Baier, Alexander S.; Pascolutti, Roberta; Wegrecki, Marcin; Zheng, Sanduo; Ong, Janice X.; Erlandson, Sarah C.; Hilger, Daniel; Rasmussen, Søren G.F.; Ring, Aaron M.; Manglik, Aashish; Kruse, Andrew C.
In: Nature Structural and Molecular Biology, Vol. 25, No. 3, 2018, p. 289-296.Research output: Contribution to journal › Journal article › Research › peer-review
Harvard
APA
Vancouver
Author
Bibtex
}
RIS
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
VL - 25
SP - 289
EP - 296
JO - Nature Structural and Molecular Biology
JF - Nature Structural and Molecular Biology
SN - 1545-9993
IS - 3
ER -
ID: 193279714