IgY14 and SuperMix immunoaffinity separations coupled with liquid chromatography–mass spectrometry for human plasma proteomics biomarker discovery

Interest in the application of advanced proteomics technologies to human blood plasma- or serum-based clinical samples for the purpose of discovering disease biomarkers continues to grow; however, the enormous dynamic range of protein concentrations in these types of samples (often >10 orders of magnitude) represents a significant analytical challenge, particularly for detecting low-abundance candidate biomarkers. In response, immunoaffinity separation methods for depleting multiple high- and moderate-abundance proteins have become key tools for enriching low-abundance proteins and enhancing detection of these proteins in plasma proteomics. Herein, we describe IgY14 and tandem IgY14-Supermix separation methods for removing 14 high-abundance and up to 60 moderate-abundance proteins, respectively, from human blood plasma and highlight their utility when combined with liquid chromatography–tandem mass spectrometry for interrogating the human plasma proteome.

► Immunoaffinity separations are key tools for enriching low-abundance proteins.
► Methods for IgY14 and SuperMix Immunoaffinity separations are described.
► IgY14 separation removes the top 14 abundant proteins from human plasma.
► Tandem IgY14-SuperMix separations remove up to 60 abundant proteins.
► Proteomics applications of IgY14 and SuperMix separations are highlighted.