Automated continuous hdx-ms device
OwnerDerek Wilson, Joseph Anacleto +1inventors
InventorsDerek Wilson, Joseph Anacleto, Cristina Lento
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Abstract
An automated continuous flow system for HDX mass spectrometry, called a continuous flow injection hydrogen deuterium exchange (CFI-HDX), or CFI-TRESI-HDX when a millisecond mixer is incorporated. In some modes, the system can also accommodate automated LC separation, improving sensitivity and allowing a wide range of buffers. Millisecond to hour HDX timepoints can be acquired continuously or discretely in a single experiment, allowing for the detection of conformational/dynamic shifts in target proteins over the full range of structural stabilities and/or binding affinities.
Claims18
A continuous flow injection time-resolved hydrogen deuterium exchange mass spectroscopy (CFI-TRESI-HDX) system, comprising:
- a first liquid chromatography (LC) pump operative to provide continuous delivery of protein samples;
- a second LC pump operative to provide continuous delivery of deuterium (D2O);
- a capillary mixer comprising an inner capillary and an outer capillary, the inner capillary coupled to the first LC pump and the outer capillary coupled to the second LC pump, the inner capillary and outer capillary in fluid communication via a notch in the inner capillary, the capillary mixer operative to produce a labeled protein sample within a labeling time between 10 milliseconds and 18 seconds, the inner capillary moveable within the outer capillary with the movement of the inner capillary providing adjustment of the labeling time;
- a quench mixer coupled to the outer capillary, the quench mixer operative to quench the labeled sample with an acid,
- wherein the output from the quench mixer is suitable for use in hydrogen-deuterium exchange mass spectroscopy.
A method of performing continuous flow injection time-resolved hydrogen deuterium exchange mass spectroscopy (CFI-TRESI-HDX), comprising:
- continuously injecting a protein sample into an inner capillary of a capillary mixer via a first liquid chromatography (LC) pump;
- continuously injecting a deuterium duffer (D2O) into an outer capillary of the capillary mixer via a second LC pump, the outer capillary and inner capillary in fluid communication via a notch in the inner capillary to produce a labeled sample;
- moving the inner capillary within the outer capillary to provide adjustment of sample labeling time within the capillary mixer to provide a labeling time between 10 milliseconds and 18 seconds;
- quenching the labeled sample in a quench mixer coupled to the outer capillary;
- and
- outputting the quenched sample for mass spectrographic analysis.
Dependencies are inferred from the claim text.