Send to

Choose Destination
J Proteome Res. 2019 Oct 4;18(10):3586-3596. doi: 10.1021/acs.jproteome.9b00254. Epub 2019 Sep 20.

Chemical Derivatization of Affinity Matrices Provides Protection from Tryptic Proteolysis.

Author information

Department of Biological Chemistry, David Geffen School of Medicine , University of California-Los Angeles , Los Angeles , California 951737 , United States.


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 spectrometry-based characterization of streptavidin-purified samples making it broadly useful for a wide variety of applications. In addition, we show that this chemical protection strategy can also be applied to other affinity matrices including immobilized antibodies against HA epitopes.


BioID; affinity purification; chemical derivatization; digestion-resistant; immunoprecipitation; mass spectrometry; proteomics; reductive methylation; streptavidin; trypsin

Supplemental Content

Full text links

Icon for American Chemical Society
Loading ...
Support Center