Enhanced performance of Tribrid Orbitrap Eclipse mass spectrometer in protein analysis

Abstract

Enhanced performance of Tribrid Orbitrap Eclipse mass spectrometer in protein analysis Jong Hwan Shin, Mi Jung Ji, SukYoun Son, Ki Soo Lee and Hyun-Mee Park Advanced Analysis Center, Korea Institute of Science and Technology For the study of omics, high accurate qualitative and quantitative analysis are required. Tribrid Orbitrap Eclipse consist of three functioning mass analyzers, the first built on quadrupole, the second on linear ion trap and, the third on orbitrap technology. Each mass analyzer must be able to work sequentially with the others which capable of performing CID, HCD or optional ETD fragmentations at any MSn stage with fragment ion detection in either ion trap or orbitrap. Specially electron transfer dissociation generates enriched peptide fragmentation for increased protein coverage and PTM(post-translation modification) identification power. We compared number of identified proteins on Eclipse orbitrap mass spectrometer and Elite orbitrap mass spectrometer. Using HeLa digested standard and nanoLC-MS/MS, we identified 3462 proteins by Eclipse and 882 proteins by Elite. Next, we comprehensively profiled glycoprotein from AGP (alpha-1-acid glycoprotein) and identified 170 glycopeptides by Eclipse and 89 glycopeptides by Elite. Using MS3 function of ion trap in Eclipse, we compared accuracies in the quantitation of TMT labeled yeast sample and Eclipse showed good accuracy. In conclusion, more accurate and sensitive detection of proteins were performed with Tribrid Orbitrap Eclipse.