Evidence map›Paper›PMID 28596009›Full record

ArticleJournal of chromatography. A2017

Subzero Celsius separations in three-zone temperature controlled hydrogen deuterium exchange mass spectrometry.

Thomas E Wales, Keith E Fadgen, Michael J Eggertson, John R Engen

Abstract read
PubMed Full text at PMC Publisher
In one paragraph

Article in Journal of chromatography. A, 2017. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 20 papers.

0numbers the graph read from it
0cells of the map it votes in
20citing papers in PubMed
–field-weighted citation impact
1 · What the graph read from it

What it found

Each row is one number read from the abstract, on the scale the paper reported it, with its interval. Left of the dashed line favours the treatment, right favours the comparator. Under each row is the sentence it came from. New to these charts? A ten-minute tutorial.

The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.

2 · The registry

The trial behind it

Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.

Neither the registry nor the abstract names a trial number. If this is a trial report, that itself is worth knowing.

3 · Its place in the literature

Who cites it

20 citing papers in PubMed.

  1. Review
  2. Article
  3. Rigorous Analysis of Multimodal HDX-MS Spectra.Journal of the American Society for Mass Spectrometry · 2025
    Article
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  5. Review
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4 · The record

Corrections and comments

PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.

5 · Who and what money

Authors and funding

4 authors.

Thomas E WalesDepartment of Chemistry & Chemical Biology, Northeastern University, Boston, MA 02115, United States.
Keith E FadgenThe Waters Corporation, Milford, MA 01757, United States.
Michael J EggertsonThe Waters Corporation, Milford, MA 01757, United States.
John R EngenDepartment of Chemistry & Chemical Biology, Northeastern University, Boston, MA 02115, United States. Electronic address: j.engen@northeastern.edu.

Funding

Extending the applications of hydrogen exchange mass spectrometryR01GM101135 · NIGMS · NORTHEASTERN UNIVERSITY · PI ENGEN, JOHN R · 2012 to 2015
$1.2M
NIGMS NIH HHS R01 GM101135
6 · The paper itself

Abstract

Hydrogen deuterium exchange mass spectrometry (HDX MS) reports on the conformational landscape of proteins by monitoring the exchange between backbone amide hydrogen atoms and deuterium in the solvent. To maintain the label for analysis, quench conditions of low temperature and pH are required during the chromatography step performed after protease digestion but before mass spectrometry. Separation at 0°C is often chosen as this is the temperature where the most deuterium can be recovered without freezing of the typical water and acetonitrile mobile phases. Several recent reports of separations at subzero Celsius emphasize the promise for retaining more deuterium and using a much longer chromatographic gradient or direct infusion time. Here we present the construction and validation of a modified Waters nanoACQUITY HDX manager with a third temperature-controlled zone for peptide separations at subzero temperatures. A new Peltier-cooled door replaces the door of a traditional main cooling chamber and the separations and trapping column are routed through the door housing. To prevent freezing, 35% methanol is introduced post online digestion. No new pumps are required and online digestion is performed as in the past. Subzero separations, using conventional HPLC column geometry of 3μ m particles in a 1×50mm column, did not result in major changes to chromatographic efficiency when lowering the temperature from 0 to -20°C. There were significant increases in deuterium recovery for both model peptides and biologically relevant protein systems. Given the higher levels of deuterium recovery, expanded gradient programs can be used to allow for higher chromatographic peak capacity and therefore the analysis of larger and more complex proteins and systems.

Indexed as

Cold TemperatureDeuterium Exchange MeasurementMass SpectrometryAmidesChemistry Techniques, AnalyticalChromatography, High Pressure LiquidDeuteriumHydrogenPeptidesProteinsSolventsAmidesDeuteriumHydrogenPeptidesProteinsSolventsDeuterium recoveryHDX MSLow temperature separationSubzero chromatography

Identifiers

PMID28596009
PMCPMC5675777

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Registered trials

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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.