Highly Specific Protein Identification by Immunoprecipitation-Mass Spectrometry Using Antifouling Microbeads

Esther Van Andel, Mark Roosjen, Stef Van Der Zanden, Stefanie C. Lange, Dolf Weijers, Maarten M.J. Smulders, Huub F.J. Savelkoul, Han Zuilhof*, Edwin J. Tijhaar

*Corresponding author for this work

Research output: Contribution to journalArticleAcademicpeer-review

3 Citations (Scopus)

Abstract

A common method to study protein complexes is immunoprecipitation (IP), followed by mass spectrometry (thus labeled: IP-MS). IP-MS has been shown to be a powerful tool to identify protein-protein interactions. It is, however, often challenging to discriminate true protein interactors from contaminating ones. Here, we describe the preparation of antifouling azide-functionalized polymer-coated beads that can be equipped with an antibody of choice via click chemistry. We show the preparation of generic immunoprecipitation beads that target the green fluorescent protein (GFP) and show how they can be used in IP-MS experiments targeting two different GFP-fusion proteins. Our antifouling beads were able to efficiently identify relevant protein-protein interactions but with a strong reduction in unwanted nonspecific protein binding compared to commercial anti-GFP beads.

Original languageEnglish
Pages (from-to)23102-23116
Number of pages15
JournalACS Applied Materials and Interfaces
Volume14
Issue number20
Early online date25 May 2022
DOIs
Publication statusPublished - 25 May 2022

Keywords

  • antibody functionalization
  • antifouling
  • click chemistry
  • immunoprecipitation
  • mass spectrometry
  • microbeads
  • proteomics
  • zwitterionic polymer brushes

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