Targeted workflows with Evosep One
Michael MacCoss (University of Washington) and Vivian Delcourt (GIE-LCH) will share how they use the Evosep One for targeted workflows and a new method, specifically designed for hydrophobic compounds, will be presented.
Using an EvoSep One for Proteomics Data Production using Data Independent Acquisition
Presenter: Michael MacCoss (Professor of Genome Sciences, University of Washington)
Our goal is to develop a high throughput method for sampling peptides with a mass spectrometer that can be used as a quantitative measure of the phenotype. To do this we have developed data independent acquisition-mass spectrometry methods that are being used for targeted analysis. That said, this does not mean that there are not substantial challenges to overcome. For example, we need methods to assess whether the peptide measurements are quantitative versus qualitative. Additionally, global methods like proteomics struggle significantly with signal calibration — making it difficult to compare quantitative measurements between batches, labs, and instrument platforms. Given the prevalence of complex proteoforms we need to think carefully about what the desired outcome is of a quantitative proteomics experiment using bottom-up methodologies. Finally, while most labs feel it is important to measure as many proteins and peptides as possible, the complications associated with doing this is non-trivial — ultimately with an increase in the number of analytes measured increases the multiple testing burden and the number of samples required to have the same statistical power. To address the challenges of running sufficient samples to maintain statistical power we have adopted the use of an EvoSep One in our workflow. The talk will present a number of diverse applications of the EvoSep One combined with our lab’s strategy for performing data independent acquisition on a quadrupole-orbitrap platform.
New standardized Evosep One gradient for ultra-sensitive doping control analysis of the most notorious synthetic anabolic steroid
Presenter: Vivian Delcourt, Ph.D. (R&D Technical Manager at GIE-LCH, Laboratoire des Courses Hippiques)
Doping control laboratories are seeking for breakthrough technologies providing improved sensitivity and consistency with robust analysis of tens of thousands samples per year. The Evosep One, designed for standardized proteomics, natively allowed the sensitive detection of protein-based banned substances such as EPO. However, to enable small-molecules analysis, a new Evosep gradient has been specifically designed for hydrophobic compounds, with the most notorious synthetic anabolic steroid “Stanozolol” as a reference. The application of such a strategy allowed the expanded detection window in complex matrices such as equine urine and hair and will constitute new landmarks for doping control laboratories.
