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Affinity-Guided Site-Selective Labeling of Nanobodies with Aldehyde Handles

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Antigen-binding proteins such as nanobodies are extensively used in biomedicine, diagnostics, and as tools for molecular biology. Often such applications require modification of the nanobodies with fluorophores or drugs. Here, we describe a robust method for introduction of aldehyde handles into His-tagged nanobodies and further derivatization of these proteins with hydroxylamine functionalized compounds of interest. The method allows for isolation of nanobodies containing one or more labels.

Original languageEnglish
Title of host publicationSingle-Domain Antibodies : Methods in Molecular Biology
EditorsGreg Hussack, Kevin A. Henry
Number of pages12
Place of publicationNew York
PublisherHumana Press
Publication year2022
ISBN (print)978-1-0716-2074-8
Publication statusPublished - 2022
SeriesMethods in Molecular Biology

Bibliographical note

© 2022. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.

    Research areas

  • Aldehyde handle, His-tag, Metal complexation, Nanobodies, Protein conjugation, Affinity Labels, Proteins/chemistry, Aldehydes/chemistry, Fluorescent Dyes, Single-Domain Antibodies/chemistry

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