Affinity Capture of Biotinylated Proteins at Acidic Conditions to Facilitate Hydrogen/Deuterium Exchange Mass Spectrometry Analysis of Multimeric Protein Complexes

Jensen, P F, et al., doi: 10.1021/ac303442y, Anal Chem, 2013

The authors present a strategy for reducing interferences during hydrogen/deuterium exchange mass spectrometry (HDX-MS) analysis of protein complexes. The method is based on streptavidin capture of biotinylated proteins under HDX-MS quench conditions. Interactions of two anti-EGFR antibodies with EGFR were studied as a model system. The Octet QK384 system was used to determine the impact of antibody biotinylation on antibody-EGFR binding. The assay showed that binding of the two biotinylated antibodies to an EGFR-Fc fusion protein loaded onto Protein G biosensors was identical to the binding of unmodified antibodies.

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