on 06-19-201701:24 PM - edited on 10-15-202106:57 AM by AnalyteGuru
Sergei I. Snovida1; Katherine E. Herting1; Ramesh Ganapathy1, Ryan Bomgarden1, Barbara J. Kaboord1, Chris Etienne1, John. C. Rogers1 ASMS 2017 Poster Introduction: The large dynamic range in protein abundance of plasma samples is the main problem associated with plasma/serum-based biomarker discovery experiments, and depletion of abundant proteins is required in order to identify and measure changes in prognostic or diagnostic plasma proteins. We have optimized the production and immobilization of immunoaffinity ligands to develop new top2 and top12 abundant protein depletion resins. In addition, we comprehensively evaluated specificity, efficiency, and reproducibility of the abundant protein depletion from human plasma samples.
Methods: A label-free approach was used for relative quantitation of human plasma proteins by LC-MS. All samples were analyzed on Thermo Scientific Orbitrap FusionTM TribridTM mass spectrometer and processed using Thermo Scientific™ Proteome Discoverer™ 1.4 and Skyline 3.6 (University of Washington) software.
Summary: We have optimized the conjugation chemistries and blending protocols to achieve >95% depletion efficiency of target high abundance proteins in human plasma samples. We have assessed depletion capacity of the new depletion resins to define the practical range of sample amounts in terms of depletion efficiencies, post-depletion protein yields, and reproducibility.