Poster 089
Improved proteomic enrichment and workflow strategies
US HUPO 2014,
Frontiers in Proteomics: Advancing Biology through Technology and Computation

Proteomic sample preparation technology for depleting hemoglobin, and enriching the low abundance proteome from human erythrocyte lysates and determination of the position and nature of human protein N termini in different tissues and disease states. Biotech Support Group has developed a series of enrichment reagents that offer common features and key advantages. After separations, the sub-proteomes retain their structural and functional integrity. All the products are consumable and economical; not being derived from biological sources. We report here applications for these products to specifically deplete albumin by avoidance strategy – called AlbuVoid™, which consequently enriches for the low abundance serum protein content. Such a strategy compares favorably to high abundance immuno-depletion (Agilent column) methods. Also, we continue to advance simplified workflows that utilize on-bead digestion for LC-MS. Together, these methods create efficiencies necessary to support high throughput investigations in all areas of discovery, targeted, functional and chemical proteomics.  

Poster: 096
Compound-Centric Displacement Proteomics – An Advantaged Method To Survey Small Molecule-Protein Interactions
US HUPO 2014,
Frontiers in Proteomics: Advancing Biology through Technology and Computation

The citation is: Abstract functional proteomics relies in part, on the functional or structural features of intact, non-denatured proteins. As such, chemical and affinity-based proteomics can be considered a subset of functional proteomics. Regardless of the evolving terminology, the subject of chemical proteomics is to identify, characterize and quantify the binding interactions of small compounds to proteomes. The consideration of those interactions as functional modulators within the cell is paramount to understanding a potential therapeutic compound’s mechanism of action. These same tools and methods also can help survey the promiscuous behavior of compounds towards multiple proteins and posit such behavior as deterministic of either toxicity or efficacy. We describe herein, new tools and methods for this purpose, called compound-centric displacement proteomics (CCDP). Employing a new product – NuGel™ PROfessor™ which can non-covalently bind proteins, a subset of proteins can be displaced upon introduction of soluble small compounds. Coupled to LC-MS, quantitative metrics of these affinity-eluted sub-proteomes help characterize and identify interacting proteins. These new methods gain efficiencies over prior covalent-based substitution methods and can serve applications in drug target deconvolution, on-target/off-target specificity, and personalized medicine.