期刊
APPLIED SCIENCES-BASEL
卷 11, 期 22, 页码 -出版社
MDPI
DOI: 10.3390/app112210883
关键词
electrospray ionisation; nanoelectrospray; proteins; alternating current; top-down
类别
资金
- Australian Research Council [DP190103298, DE190100986, FT200100798]
- Australian Research Council [DE190100986, FT200100798] Funding Source: Australian Research Council
Pulsed nanoESI-MS using nanoscale emitters can improve the performance of intact protein detection, with increased ion abundances and decreased noise compared to conventional methods. Optimal repetition rates significantly enhance the signal levels for proteins.
Electrospray ionisation (ESI) is renowned for its ability to ionise intact proteins for sensitive detection by mass spectrometry (MS). However, the use of a conventional direct current ESI voltage can result in the formation of relatively large initial droplet sizes, which can limit efficient ion desolvation and sensitivity. Here, pulsed nanoESI (nESI) MS using nanoscale emitters with inner diameters of ~250 nm is reported. In this approach, the nESI voltage is rapidly pulsed from 0 to ~1.5 kV with sub-nanosecond rise times, duty cycles from 10 to 90%, and repetition rates of 10 to 350 kHz. Using pulsed nESI, the performance of MS for the detection of intact proteins can be improved in terms of increased ion abundances and decreased noise. The absolute ion abundances and signal-to-noise levels of protonated ubiquitin, cytochrome C, myoglobin, and carbonic anhydrase II formed from standard denaturing solutions can be increased by up to 82% and 154% using an optimal repetition rate of ~200 kHz compared to conventional nESI-MS. Applying pulsed nESI-MS to a mixture of four proteins resulted in the signal for each protein increasing by up to 184% compared to the more conventional nESI-MS. For smaller ions (& LE;1032 m/z), the signal can also be increased by the use of high repetition rates (200-250 kHz), which is consistent with the enhanced performance depending more on general factors associated with the ESI process (e.g., smaller initial droplet sizes and reduced Coulombic repulsion in the spray plume) rather than analyte-specific effects (e.g., electrophoretic mobility). The enhanced sensitivity of pulsed nESI is anticipated to be beneficial for many different types of tandem mass spectrometry measurements.
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