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PXD016435

PXD016435 is an original dataset announced via ProteomeXchange.

Dataset Summary
TitleShotgun proteomics analysis of protein purifications from human plasma using antibodies and affimers
DescriptionAntibodies are widely used to purify protein from complex mixtures like human plasma. Assessing the specificity of anntibodies, or any protein binder, is essential for the development of a protein biomarker assay. In addition to antibodies, novel protein binders like affimers are developed with a high specificity against the target protein and without batch-to-batch variability. In this study, we compared the specificity of affimers and commercially available antibodies against two non-related proteins (IL-37 and proinsulin). We conducted protein capture experiments with antibodies coupled to magnetic protein G beads and (biotinylated) affimers coupled to streptavidin magnetic beads. Binding capacity of anti-IL-37 and anti-proinsulin antibodies and affimers was investigated via these magnetic-bead captures of their recombinant protein targets in human plasma. The purified fractions were transferred to an SDS-PAGE, followed by in-gel digestion of the visualized protein bands and subsequently analyzed with western blot or shotgun proteomics using LC-MS/MS. From the proteomics experiments we evaluated the amount of background proteins that were co-purified during the protein purification procedure. In addition we determined the relative abundance of the target protein relative to other non-related proteins.
HostingRepositoryPRIDE
AnnounceDate2024-10-22
AnnouncementXMLSubmission_2024-10-22_05:06:41.635.xml
DigitalObjectIdentifier
ReviewLevelPeer-reviewed dataset
DatasetOriginOriginal dataset
RepositorySupportUnsupported dataset by repository
PrimarySubmitterRoel Tans
SpeciesList scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationListmonohydroxylated residue; acetylated residue; iodoacetamide derivatized residue
InstrumentmaXis
Dataset History
RevisionDatetimeStatusChangeLog Entry
02019-11-25 07:41:54ID requested
12020-06-02 02:02:04announced
22024-10-22 05:06:42announced2024-10-22: Updated project metadata.
Publication List
10.1016/j.pep.2020.105677;
Tans R, van Rijswijck DMH, Davidson A, Hannam R, Ricketts B, Tack CJ, Wessels HJCT, Gloerich J, van Gool AJ, Affimers as an alternative to antibodies for protein biomarker enrichment. Protein Expr Purif, 174():105677(2020) [pubmed]
Keyword List
submitter keyword: antibody,protein purification, affimer, LC-MS/MS
Contact List
Alain van Gool
contact affiliationTranslational Metabolic Laboratory, department of Laboratory Medicine. Radboud institute for molecular life sciences, Radboud University Medical Center, Nijmegen, the Netherlands
contact emailalain.vangool@radboudumc.nl
lab head
Roel Tans
contact affiliationRadboudumc
contact emailroeltans@outlook.com
dataset submitter
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Dataset FTP location
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