A general protocol for the generation of Nanobodies for structural biology

Research output: Contribution to journal › Journal article › Research › peer-review

Standard

A general protocol for the generation of Nanobodies for structural biology. / Pardon, Els; Laeremans, Toon; Triest, Sarah; Rasmussen, Søren Gøgsig Faarup; Wohlkönig, Alexandre; Ruf, Armin; Muyldermans, Serge; Hol, Wim G J; Kobilka, Brian K; Steyaert, Jan.

In: Nature Protocols (Print), Vol. 9, No. 3, 03.2014, p. 674-93.

Research output: Contribution to journal › Journal article › Research › peer-review

Harvard

Pardon, E, Laeremans, T, Triest, S, Rasmussen, SGF, Wohlkönig, A, Ruf, A, Muyldermans, S, Hol, WGJ, Kobilka, BK & Steyaert, J 2014, 'A general protocol for the generation of Nanobodies for structural biology', Nature Protocols (Print), vol. 9, no. 3, pp. 674-93. https://doi.org/10.1038/nprot.2014.039

APA

Pardon, E., Laeremans, T., Triest, S., Rasmussen, S. G. F., Wohlkönig, A., Ruf, A., Muyldermans, S., Hol, W. G. J., Kobilka, B. K., & Steyaert, J. (2014). A general protocol for the generation of Nanobodies for structural biology. Nature Protocols (Print), 9(3), 674-93. https://doi.org/10.1038/nprot.2014.039

Vancouver

Pardon E, Laeremans T, Triest S, Rasmussen SGF, Wohlkönig A, Ruf A et al. A general protocol for the generation of Nanobodies for structural biology. Nature Protocols (Print). 2014 Mar;9(3):674-93. https://doi.org/10.1038/nprot.2014.039

Author

Pardon, Els ; Laeremans, Toon ; Triest, Sarah ; Rasmussen, Søren Gøgsig Faarup ; Wohlkönig, Alexandre ; Ruf, Armin ; Muyldermans, Serge ; Hol, Wim G J ; Kobilka, Brian K ; Steyaert, Jan. / A general protocol for the generation of Nanobodies for structural biology. In: Nature Protocols (Print). 2014 ; Vol. 9, No. 3. pp. 674-93.

Bibtex

@article{cf2c082cbdea47bca77489dc0b49e16e,
title = "A general protocol for the generation of Nanobodies for structural biology",
abstract = "There is growing interest in using antibodies as auxiliary tools to crystallize proteins. Here we describe a general protocol for the generation of Nanobodies to be used as crystallization chaperones for the structural investigation of diverse conformational states of flexible (membrane) proteins and complexes thereof. Our technology has a competitive advantage over other recombinant crystallization chaperones in that we fully exploit the natural humoral response against native antigens. Accordingly, we provide detailed protocols for the immunization with native proteins and for the selection by phage display of in vivo-matured Nanobodies that bind conformational epitopes of functional proteins. Three representative examples illustrate that the outlined procedures are robust, making it possible to solve by Nanobody-assisted X-ray crystallography in a time span of 6-12 months.",
author = "Els Pardon and Toon Laeremans and Sarah Triest and Rasmussen, {S{\o}ren G{\o}gsig Faarup} and Alexandre Wohlk{\"o}nig and Armin Ruf and Serge Muyldermans and Hol, {Wim G J} and Kobilka, {Brian K} and Jan Steyaert",
year = "2014",
month = mar,
doi = "10.1038/nprot.2014.039",
language = "English",
volume = "9",
pages = "674--93",
journal = "Nature Protocols",
issn = "1754-2189",
publisher = "nature publishing group",
number = "3",

}

RIS

TY - JOUR

T1 - A general protocol for the generation of Nanobodies for structural biology

AU - Pardon, Els

AU - Laeremans, Toon

AU - Triest, Sarah

AU - Rasmussen, Søren Gøgsig Faarup

AU - Wohlkönig, Alexandre

AU - Ruf, Armin

AU - Muyldermans, Serge

AU - Hol, Wim G J

AU - Kobilka, Brian K

AU - Steyaert, Jan

PY - 2014/3

Y1 - 2014/3

N2 - There is growing interest in using antibodies as auxiliary tools to crystallize proteins. Here we describe a general protocol for the generation of Nanobodies to be used as crystallization chaperones for the structural investigation of diverse conformational states of flexible (membrane) proteins and complexes thereof. Our technology has a competitive advantage over other recombinant crystallization chaperones in that we fully exploit the natural humoral response against native antigens. Accordingly, we provide detailed protocols for the immunization with native proteins and for the selection by phage display of in vivo-matured Nanobodies that bind conformational epitopes of functional proteins. Three representative examples illustrate that the outlined procedures are robust, making it possible to solve by Nanobody-assisted X-ray crystallography in a time span of 6-12 months.

AB - There is growing interest in using antibodies as auxiliary tools to crystallize proteins. Here we describe a general protocol for the generation of Nanobodies to be used as crystallization chaperones for the structural investigation of diverse conformational states of flexible (membrane) proteins and complexes thereof. Our technology has a competitive advantage over other recombinant crystallization chaperones in that we fully exploit the natural humoral response against native antigens. Accordingly, we provide detailed protocols for the immunization with native proteins and for the selection by phage display of in vivo-matured Nanobodies that bind conformational epitopes of functional proteins. Three representative examples illustrate that the outlined procedures are robust, making it possible to solve by Nanobody-assisted X-ray crystallography in a time span of 6-12 months.

U2 - 10.1038/nprot.2014.039

DO - 10.1038/nprot.2014.039

M3 - Journal article

C2 - 24577359

VL - 9

SP - 674

EP - 693

JO - Nature Protocols

JF - Nature Protocols

SN - 1754-2189

IS - 3

ER -

ID: 120587145