High‐Throughput Equine Doping Controls on a Trapped Ion Mobility Quadrupole‐Time‐of‐Flight Mass Spectrometer: Technical Considerations of dia/slice/prmPASEF Applied to the Long‐Term Detection of Monoclonal Antibodies

Abstract

Data‐independent acquisition (DIA) methods employing a scanning quadrupole were recently described across multiple platforms. These strategies display remarkable performances in untargeted proteomics studies thanks to rapid duty cycles, leading to ultrashort liquid chromatography gradients while maintaining enough data points per peaks when coupled to fast‐scanning mass analyzer. In this article, we perform the evaluation of three data acquisition strategies named diaPASEF,slicePASEF, and prmPASEF on a trapped ion mobility spectrometry quadrupole‐time‐of‐flight (TIMS‐Q‐TOF) mass spectrometer for high‐throughput doping control screening analyses. We report that slicePASEF outperforms diaPASEF and is almost as sensitive as prmPASEF in detecting humanized monoclonal antibodies for several weeks in equine plasma after administration. We observed that diaPASEF is still providing the best performances in untargeted proteomics studies employing high amounts of input materials, which is linked with the high complexity of slicePASEF data and current processing algorithms.