Yeast surface display platform for rapid discovery of conformationally selective nanobodies

Conor McMahon; Alexander S. Baier; Roberta Pascolutti; Marcin Wegrecki; Sanduo Zheng; Janice X. Ong; Sarah C. Erlandson; Daniel Hilger; Søren G.F. Rasmussen; Aaron M. Ring; Aashish Manglik; Andrew C. Kruse

doi:10.1038/s41594-018-0028-6

Yeast surface display platform for rapid discovery of conformationally selective nanobodies

Conor McMahon, Alexander S. Baier, Roberta Pascolutti, Marcin Wegrecki, Sanduo Zheng, Janice X. Ong, Sarah C. Erlandson, Daniel Hilger, Søren G.F. Rasmussen, Aaron M. Ring, Aashish Manglik^*, Andrew C. Kruse

^*Corresponding author for this work

Neuropharm and Genetics

112 Citations (Scopus)

Abstract

Camelid single-domain antibody fragments ('nanobodies') provide the remarkable specificity of antibodies within a single 15-kDa immunoglobulin V_HH 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.

Original language	English
Journal	Nature Structural and Molecular Biology
Volume	25
Issue number	3
Pages (from-to)	289-296
Number of pages	8
ISSN	1545-9993
DOIs	https://doi.org/10.1038/s41594-018-0028-6
Publication status	Published - 2018

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McMahon, C., Baier, A. S., Pascolutti, R., Wegrecki, M., Zheng, S., Ong, J. X., Erlandson, S. C., Hilger, D., Rasmussen, S. G. F., Ring, A. M., Manglik, A., & Kruse, A. C. (2018). Yeast surface display platform for rapid discovery of conformationally selective nanobodies. Nature Structural and Molecular Biology, 25(3), 289-296. https://doi.org/10.1038/s41594-018-0028-6

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title = "Yeast surface display platform for rapid discovery of conformationally selective nanobodies",

abstract = "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.",

author = "Conor McMahon and Baier, {Alexander S.} and Roberta Pascolutti and Marcin Wegrecki and Sanduo Zheng and Ong, {Janice X.} and Erlandson, {Sarah C.} and Daniel Hilger and Rasmussen, {S{\o}ren G.F.} and Ring, {Aaron M.} and Aashish Manglik and Kruse, {Andrew C.}",

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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

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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.

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DO - 10.1038/s41594-018-0028-6

M3 - Journal article

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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 -

Yeast surface display platform for rapid discovery of conformationally selective nanobodies

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