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

Anders Märcher, Kurt V. Gothelf*

*Corresponding author for this work

Research output: Contribution to book/anthology/report/proceedingBook chapterResearchpeer-review

Abstract

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
Volume2446
Place of publicationNew York
PublisherHumana Press
Publication date2022
Pages345-356
ISBN (Print)978-1-0716-2074-8
DOIs
Publication statusPublished - 2022
SeriesMethods in Molecular Biology
Volume2446
ISSN1064-3745

Keywords

  • 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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