Chemical Derivatization of Affinity Matrices Provides Protection from Tryptic Proteolysis.

ABSTRACT: The enrichment of biotinylated proteins using immobilized streptavidin has become a staple methodology for affinity purification-based proteomics. Many of these workflows rely upon tryptic digestion to elute streptavidin-captured moieties from the beads. The concurrent release of high amounts of streptavidin-derived peptides into the digested sample, however, can significantly hamper the effectiveness of downstream proteomic analyses by increasing the complexity and dynamic range of the mixture. Here, we describe a strategy for the chemical derivatization of streptavidin that renders it largely resistant to proteolysis by trypsin and thereby dramatically reduces the amount of streptavidin contamination in the sample. This rapid and robust approach improves the effectiveness of mass spectro

SUBMITTER: Barshop WD 

PROVIDER: S-EPMC6872193 | biostudies-literature | 2019 Oct

REPOSITORIES: biostudies-literature