on 06-13-201901:09 PM - edited on 10-15-202110:51 AM by Closed Account
Jing Wang1; Sarah Trusiak1; Ryan D. Bomgarden2; Sergei Snovida2; Emily I. Chen1 ASMS 2019 Poster Purpose – This poster presents a high throughput serum and plasma proteomics analysis workflow for large population cohorts.
Methods – To reduce analytical variability of plasma sample preparation for LC-MS/MS analysis, we automated Thermo Scientific™ EasyPep™ Mini MS Sample Prep Kit using the Hamilton Microlab STARlet liquid handling system with [MPE]2 positive pressure and evaporation modules. The Evosep LC system was used to run high throughput and automated LC methods. Thermo Scientific™ Q Exactive™ HF-X MS and data-dependent acquisition (DDA) were used to generate quantitative LFQ plasma proteome data. Skyline was used for retention time analysis and Thermo Scientific™ Proteome Discoverer™ 2.3 software was used for database search and post-data analysis.
Results – The automated sample preparation can process 96 samples within 4 hours with ~80% recovery. The throughput of LC analysis for the standardized workflow is > 50 samples per day with 10% overhead to minimize sample carrying over. Around 150 and 200 core proteins (high confident) could be reproducibly identified and quantified for the undepleted serum and the depleted plasma samples, respectively. An example of this workflow applied to small scale depleted plasma lung cancer samples is presented in this poster as well.
1Thermo Fisher Scientific Precision Medicine Science Center, Cambridge, MA
2ThermoFisher Scientific, Rockford, IL