PXD063440-1

PXD063440 is an original dataset announced via ProteomeXchange.

Title	Native Taylor/Non-Taylor Dispersion – Mass Spectrometry (TNT-MS) Allows Rapid Protein Desalting and Multiplexed, Label-Free Ligand Screening
Description	Native mass spectrometry (MS) has become an important technique in several fields including structural biology and drug discovery, due to its ability to study non-covalent assemblies in the gas phase. In many settings, the main drawbacks of native MS are the incompatibility of electrospray ionisation with non-volatile salts and the risk of protein signal suppression if small, efficiently ionising molecules are present in the sample. This often requires an offline buffer exchange step and/or parallel sample preparation workflow to other analytical methods, reducing both the adoption and the throughput of native MS. Here, we exploit the dynamics of analytes flowing through an open tubular capillary to keep molecules with a small hydrodynamic radius (e.g., salts) inside a Taylor dispersion regime, while pushing larger species (e.g., proteins) into a non-Taylor regime. The result of this is that the larger species elute earlier, and are effectively buffer exchanged within the capillary on a timescale of approximately 30 seconds. In addition to desalting of proteins injected in solutions containing 25 to 200 mM NaCl and other biologically relevant buffers (e.g., HEPES, TCEP, and glycerol), we also demonstrate that this method can separate unbound small molecules from protein-ligand complexes, enabling rapid, multiplexed ligand screening based on native MS. Finally, we investigated the dependence of the critical flow rate required to push proteins outside the Taylor regime on protein size, enabling limited size-based separation of proteins and providing a starting point for others to adopt this method. Taylor/non-Taylor dispersion mass spectrometry (TNT-MS) was implemented using an unmodified LC-MS system operated without a chromatographic column and coupled to an autosampler. This allows significant automation, which we believe will contribute to the wider adoption of native MS as a routine method.
HostingRepository	PRIDE
AnnounceDate	2025-07-10
AnnouncementXML	Submission_2025-07-10_00:51:54.263.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Jonathan Eisert
SpeciesList	scientific name: Mus musculus (Mouse); NCBI TaxID: 10090; scientific name: Bos taurus (Bovine); NCBI TaxID: 9913; scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationList	No PTMs are included in the dataset
Instrument	Synapt MS

Revision	Datetime	Status	ChangeLog Entry
0	2025-04-29 05:05:36	ID requested
⏵ 1	2025-07-10 00:51:54	announced

10.1002/SMTD.202500658;

submitter keyword: Ligand screening,native MS, Taylor dispersion

Prof. Frederik Lermyte
contact affiliation	Department of Chemistry, Clemens-Schöpf-Institute of Chemistry and Biochemistry, Technical University of Darmstadt, Peter-Grünberg-Strasse 4, 64287 Darmstadt, Germany
contact email	frederik.lermyte@tu-darmstadt.de
lab head
Jonathan Eisert
contact affiliation	Department of Chemistry, Clemens-Schöpf-Institute of Chemistry and Biochemistry, Technical University of Darmstadt, Peter-Grünberg-Strasse 4, 64287 Darmstadt, Germany
contact email	jonathan.eisert@tu-darmstadt.de
dataset submitter

Dataset FTP location NOTE: Most web browsers have now discontinued native support for FTP access within the browser window. But you can usually install another FTP app (we recommend FileZilla) and configure your browser to launch the external application when you click on this FTP link. Or otherwise, launch an app that supports FTP (like FileZilla) and use this address: ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2025/07/PXD063440

PRIDE project URI

• PRIDE
  1. PXD063440
     1. Label: PRIDE project
     2. Name: Native Taylor/Non-Taylor Dispersion – Mass Spectrometry (TNT-MS) Allows Rapid Protein Desalting and Multiplexed, Label-Free Ligand Screening