PXD026637-1

PXD026637 is an original dataset announced via ProteomeXchange.

Title	Getting more out of co-immunoprecipitation mass spectrometry experiments by reducing interference using FAIMS
Description	Co-immunoprecipitation of proteins coupled to mass spectrometry has transformed modern biology’s understanding of protein interaction networks. These approaches exploit the selective isolation of tagged proteins by affinity enrichment / purification to identify protein binding partners at scale and in an unbiased manner. In instances where a suitable antibody is not be available it is common to graft synthetic tags such as FLAG or His Tags onto target protein sequences allowing the use of commercially available and validated antibodies for affinity purification. To allow the selective elution of protein complexes competitive displacement using a large molar excess of the tag peptide is widely used. Yet, this creates downstream challenges for the mass spectrometry analysis due to the presence of large quantities of a contaminating peptide. Here, we demonstrate that Field Asymmetric Ion Mobility (FAIMS), a gas phase ion separation device can be applied to FLAG-tagged and His Tag pull down assay to increase the depth of protein coverage in these experiments. By excluding tag peptides based on their ion mobility profiles we demonstrate that single compensation voltage, or stepped compensation voltages strategies can significantly increase the coverage of total proteins by up to 2.5-folds and unique proteins by up to 15-folds versus experiments that do not use FAIMS. Combined these results highlight FAIMS is able to improve proteome depth by excluding interfering peptides without the need for additional sample handling or altering sample preparation protocols.
HostingRepository	PRIDE
AnnounceDate	2025-10-16
AnnouncementXML	Submission_2025-10-15_16:20:27.463.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Ching-Seng Ang
SpeciesList	scientific name: Rabies virus Nishigahara RCEH; NCBI TaxID: NEWT:11298; scientific name: Homo sapiens (Human); NCBI TaxID: NEWT:9606;
ModificationList	acetylated residue; monohydroxylated residue; iodoacetamide derivatized residue
Instrument	Orbitrap Eclipse; Orbitrap Fusion Lumos; Vion IMS QTof; Orbitrap Exploris 480

Revision	Datetime	Status	ChangeLog Entry
0	2021-06-10 22:47:58	ID requested
⏵ 1	2025-10-15 16:20:28	announced

Ang CS, Sacharz J, Leeming MG, Nie S, Varshney S, Scott NE, Williamson NA, Getting more out of FLAG-Tag co-immunoprecipitation mass spectrometry experiments using FAIMS. J Proteomics, 254():104473(2022) [pubmed]

10.1016/j.jprot.2021.104473;

submitter keyword: FAIMS,Co-IP mass spectrometry, Proteomics, ion mobility

Nicholas A. Williamson
contact affiliation	Bio21 Mass Spectrometry and Proteomics Facility, The University of Melbourne, Parkville, Victoria 3010, Australia
contact email	nawill@unimelb.edu.au
lab head
Ching-Seng Ang
contact affiliation	University of Melbourne
contact email	cang@unimelb.edu.au
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD026637
     1. Label: PRIDE project
     2. Name: Getting more out of co-immunoprecipitation mass spectrometry experiments by reducing interference using FAIMS