Dynamics of ions of intact proteins in the Orbitrap mass analyzer.
While allowing analysis of intact proteins without a theoretical upper mass limit, the Orbitrap mass analyzer demonstrates reduced resolving power as ion mass increases even at a constant mass-to-charge ratio. It is shown that this effect comes from the effects of ion scattering on background gas molecules. The main mechanisms causing decay of acquired transient appear to be fragmentation as well as accelerated dephasing of ion packets. Isotopic resolution of proteins including bovine serum albumin (MW 66.4 kDa) and transferrin (MW 78 kDa) has also been demonstrated. As a part of this study, detection of individual multiply-charged ions of myoglobin (MW 16.9 kDa) has been demonstrated. Quantized distribution of signal intensities for +20 myoglobin ions well above the noise threshold was observed, with high mass accuracy and resolution of recorded individual ions used as an independent confirmation of correct assignment of signal to ions rather than to noise. The latter also allowed us to benchmark the sensitivity of image-current detection and explore in detail factors responsible for signal decay.
Makarov
Alexander
A
Thermo Fisher Scientific, Bremen, Germany. alexander.makarov@thermofisher.com
Denisov
Eduard
E
D003198:Computer Simulation; D017076:Computer-Aided Design; D004867:Equipment Design; D019544:Equipment Failure Analysis; D007477:Ions; D008956:Models, Chemical; D011506:Proteins / Q000032:analysis* / Q000737:chemistry*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D021241:Spectrometry, Mass, Electrospray Ionization / Q000295:instrumentation* / Q000379:methods*
D007477:Ions; D011506:Proteins