PXD016435

PXD016435 is an original dataset announced via ProteomeXchange.

Title	Shotgun proteomics analysis of protein purifications from human plasma using antibodies and affimers
Description	Antibodies 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.
HostingRepository	PRIDE
AnnounceDate	2024-10-22
AnnouncementXML	Submission_2024-10-22_05:06:41.635.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Roel Tans
SpeciesList	scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationList	monohydroxylated residue; acetylated residue; iodoacetamide derivatized residue
Instrument	maXis

Revision	Datetime	Status	ChangeLog Entry
0	2019-11-25 07:41:54	ID requested
1	2020-06-02 02:02:04	announced
⏵ 2	2024-10-22 05:06:42	announced	2024-10-22: Updated project metadata.

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]

submitter keyword: antibody,protein purification, affimer, LC-MS/MS

Alain van Gool
contact affiliation	Translational Metabolic Laboratory, department of Laboratory Medicine. Radboud institute for molecular life sciences, Radboud University Medical Center, Nijmegen, the Netherlands
contact email	alain.vangool@radboudumc.nl
lab head
Roel Tans
contact affiliation	Radboudumc
contact email	roeltans@outlook.com
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/2020/05/PXD016435

PRIDE project URI

• PRIDE
  1. PXD016435
     1. Label: PRIDE project
     2. Name: Shotgun proteomics analysis of protein purifications from human plasma using antibodies and affimers