PXD043709 is an
original dataset announced via ProteomeXchange.
Dataset Summary
Title | Chemical acetylation of ligands and two-step digestion protocol for reducing co-digestion in affinity purification-mass spectrometry |
Description | We present an effective, fast and user-friendly method to reduce co-digestion of bead-bound ligands, such as antibodies or streptavidin, in affinity purification-mass spectrometry experiments. A short pre-incubation of beads with Sulfo-NHS-Acetate leads to chemical acetylation of lysine residues, making ligands insusceptible to Lys-C-mediated proteolysis. In contrast to similar approaches, our procedure offers the advantage of exclusively using non-toxic chemicals and employing mild chemical reaction conditions. After binding of bait proteins to Sulfo-NHS-Acetate treated beads, we employ a two-step digestion protocol with the sequential use of Lys-C protease for on-bead digestion followed by in-solution digestion of the released proteins with trypsin. The implementation of this protocol results in a strong reduction of contaminating ligand peptides, which allows significantly higher amounts of sample to be subjected to LC-MS analysis, improving sensitivity and quantitative accuracy. |
HostingRepository | PRIDE |
AnnounceDate | 2023-11-14 |
AnnouncementXML | Submission_2023-11-14_09:06:10.595.xml |
DigitalObjectIdentifier | |
ReviewLevel | Peer-reviewed dataset |
DatasetOrigin | Original dataset |
RepositorySupport | Unsupported dataset by repository |
PrimarySubmitter | Wolfgang Reiter |
SpeciesList | scientific name: Homo sapiens (Human); NCBI TaxID: 9606; scientific name: Saccharomyces cerevisiae (Baker's yeast); NCBI TaxID: 4932; |
ModificationList | No PTMs are included in the dataset |
Instrument | Q Exactive HF-X |
Dataset History
Revision | Datetime | Status | ChangeLog Entry |
0 | 2023-07-12 03:50:23 | ID requested | |
1 | 2023-10-23 05:15:44 | announced | |
⏵ 2 | 2023-11-14 09:06:16 | announced | 2023-11-14: Updated project metadata. |
Publication List
Bordons M, Zulueta MA, [Evaluation of the scientific activity through bibliometric indices]. Rev Esp Cardiol, 52(10):790-800(1999) [pubmed] |
Keyword List
submitter keyword: chemical acetylation, immunopurification, streptavidin, ligand,affinity purification, Sulfo-NHS-Acetate, nanobody, GFP |
Contact List
Markus Hartl |
contact affiliation | Mass Spectrometry Facility, Max Perutz Labs Vienna, University of Vienna, Vienna Biocenter, Dr.-Bohr-Gasse 7, 1030 Vienna |
contact email | markus.hartl@univie.ac.at |
lab head | |
Wolfgang Reiter |
contact affiliation | Mass Spectrometry Facility, Max Perutz Labs Vienna, University of Vienna, Vienna Biocenter, Dr.-Bohr-Gasse 7, 1030 Vienna |
contact email | wolfgang.l.reiter@univie.ac.at |
dataset submitter | |
Full Dataset Link List
Dataset FTP location
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PRIDE project URI |
Repository Record List
[ + ]
[ - ]
- PRIDE
- PXD043709
- Label: PRIDE project
- Name: Chemical acetylation of ligands and two-step digestion protocol for reducing co-digestion in affinity purification-mass spectrometry