Supercharging and Multiple Reaction Monitoring Optimization of High Molecular Weight Intact Proteins using Triple Quadrupole Mass Spectrometry
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Date
2019-07-17Author
Khanal, Durga Devi
0000-0003-0803-1621
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Various supercharging agents, such as meta-nitrobenzyl alcohol, dimethyl sulfoxide, trifluoroethanol (TFE), and sulfolane were tested in different concentrations to study the effects of supercharging agents on the charge distributions of the high molecular weight intact proteins. The data were analyzed by triple quadrupole mass spectrometry. For most of the proteins, m-NBA and TFE worked more effectively than other supercharging agents, both to shift the charge state and increase intensity. The settings of different instrumental parameters were also studied. Both increases in source temperature and flow rate had a negative impact on average charge state observed. The goal of this work was to increase charge acquisition and ionization efficiency for proteins ranging from 66 kDa – 150 kDa, to allow subsequent optimization of multiple reaction monitoring (MRM) mode transitions for potential top-down quantitative analysis. The MRM transitions of six to eight different precursor ions of the proteins were optimized to demonstrate the feasibility of this approach and limits of detection were evaluated.