Apparatus for Automated Continuous Hydrogen Deuterium Exchange Mass Spectrometry Measurements from Milliseconds to Hours

Hydrogen deuterium
exchange mass spectrometry (HDX-MS) is a rapidly
growing technique for protein characterization in industry and academia,
complementing the “static” picture provided by classical
structural biology with information about the dynamic structural changes
that accompany biological function. Conventional hydrogen deuterium
exchange experiments, carried out on commercially available systems,
typically collect 4–5 exchange timepoints on a timescale ranging
from tens of seconds to hours using a workflow that can require 24
h or more of continuous data collection for triplicate measurements.
A small number of groups have developed setups for millisecond timescale
HDX, allowing for the characterization of dynamic shifts in weakly
structured or disordered regions of proteins. This capability is particularly
important given the central role that weakly ordered protein regions
often play in protein function and pathogenesis. In this work, we
introduce a new continuous flow injection setup for time-resolved
HDX-MS (CFI-TRESI-HDX) that allows automated, continuous or discrete
labeling time measurements from milliseconds to hours. The device
is composed almost entirely of “off-the-shelf” LC components
and can acquire an essentially unlimited number of timepoints with
substantially reduced runtimes compared to conventional systems.