An Improved Rapid Mixing Device for Time-Resolved Electrospray Mass Spectrometry Measurements |
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Authors: | Nicholas Zinck Ann-Kathrin Stark Derek J Wilson Michal Sharon |
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Affiliation: | [a]Department of Biological Chemistry, Weizmann Institute of Science, 76100 Rehovot (Israel), E-mail: ;[b]Department of Chemistry, York University, Toronto, ON M3J 1P3 (Canada) |
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Abstract: | Time series data can provide valuable insight into the complexity of biological reactions. Such information can be obtained by mass-spectrometry-based approaches that measure pre-steady-state kinetics. These methods are based on a mixing device that rapidly mixes the reactants prior to the on-line mass measurement of the transient intermediate steps. Here, we describe an improved continuous-flow mixing apparatus for real-time electrospray mass spectrometry measurements. Our setup was designed to minimize metal–solution interfaces and provide a sheath flow of nitrogen gas for generating stable and continuous spray that consequently enhances the signal-to-noise ratio. Moreover, the device was planned to enable easy mounting onto a mass spectrometer replacing the commercial electrospray ionization source. We demonstrate the performance of our apparatus by monitoring the unfolding reaction of cytochrome C, yielding improved signal-to-noise ratio and reduced experimental repeat errors. |
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Keywords: | kinetics mass spectrometry on-line measurements proteins rapid mixing device time-resolved experiments structure characterization of biomolecules |
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