@inbook{cdc85d34dd8146b9bc60d772f11e82b4,
title = "Introducing Cysteines into Nanobodies for Site-Specific Labeling",
abstract = "We have developed a generally applicable methodology for cysteine mutagenesis of nanobody (Nb) framework region serine residues. This strategy allows for subsequent labeling with thiol-reactive compounds without disrupting Nb antigen binding. We provide a protocol for production, labeling, and affinity determination of cysteine-engineered Nbs (cys-Nbs) with Alexa Fluor 488-maleimide and the mercury compound para-chloromercuribenzoic acid (PCMB). Alexa Fluor 488- and PCMB-labeled cys-Nbs can be used for immunofluorescence microscopy and experimental phasing in crystallography, respectively.",
keywords = "Cysteine, Experimental phasing, Imaging, Maleimide, Mercury, Nanobody, Site-specific labeling",
author = "Hansen, {Simon Boje} and Andersen, {Kasper R{\o}jkj{\ae}r}",
note = "Publisher Copyright: {\textcopyright} 2022, The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.",
year = "2022",
doi = "10.1007/978-1-0716-2075-5_16",
language = "English",
isbn = "978-1-0716-2074-8 ",
series = "Methods in Molecular Biology",
publisher = "Humana Press",
pages = "327--343",
editor = "Hussack, {Greg } and Henry, {Kevin A. }",
booktitle = "Single-Domain Antibodies",
}