Open AccessHighly Specific Protein Identification by Immunoprecipitation–Mass Spectrometry Using Antifouling Microbeads
Author(s) -
Esther van Andel,
Mark Roosjen,
Stef van der Zanden,
Stefanie Lange,
Dolf Weijers,
Maarten M. J. Smulders,
H.F.J. Savelkoul,
Han Zuilhof,
Edwin Tijhaar
Publication year - 2022
Publication title -
acs applied materials and interfaces
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.535
H-Index - 228
eISSN - 1944-8252
pISSN - 1944-8244
DOI - 10.1021/acsami.1c22734
Subject(s) - immunoprecipitation , green fluorescent protein , biofouling , mass spectrometry , fusion protein , protein–protein interaction , fluorescence , biophysics , chemistry , chromatography , combinatorial chemistry , nanotechnology , materials science , biochemistry , biology , gene , recombinant dna , physics , quantum mechanics , membrane
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.