Nicholas M. Riley, Christopher Mullen, Chad R. Weisbrod, Seema Sharma, Michael W. Senko, Vlad Zabrouskov, Michael S. Westphall, John E. P. Syka, Joshua J. Coon
J. Am. Soc. Mass Spectrom. (2015)
Electron transfer dissociation (ETD) is a valuable tool for protein sequence analysis, especially for the fragmentation of intact proteins. However, low product ion signal-to-noise often requires some degree of signal averaging to achieve high quality MS/MS spectra of intact proteins. Here we describe a new implementation of ETD on the newest generation of quadrupole-Orbitrap-linear ion trap Tribrid, the Orbitrap Fusion Lumos, for improved product ion signal-to-noise via ETD reactions on larger precursor populations. In this new high precursor capacity ETD implementation, precursor cations are accumulated in the center section of the high pressure cell in the dual pressure linear ion trap prior to charge-sign independent trapping, rather than precursor ion sequestration in only the back section as is done for standard ETD. This new scheme increases the charge capacity of the precursor accumulation event, enabling storage of approximately 3-fold more precursor charges. High capacity ETD boosts the number of matching fragments identified in a single MS/MS event, reducing the need for spectral averaging. These improvements in intra-scan dynamic range via reaction of larger precursor populations, which have been previously demonstrated through custom modified hardware, are now available on a commercial platform, offering considerable benefits for intact protein analysis and top down proteomics. In this work, we characterize the advantages of high precursor capacity ETD through studies with myoglobin and carbonic anhydrase.
Department of Chemistry, University of Wisconsin-Madison, Madison, WI 53706, USA Thermo Fisher Scientific, San Jose, CA 95134, USA Department of Biomolecular Chemistry, University of Wisconsin-Madison, Madison, WI 53706, USA Genome Center of Wisconsin, University of Wisconsin-Madison, Madison, WI 53706, USA