⮝ Full datasets listing

PXD017316

PXD017316 is an original dataset announced via ProteomeXchange.

Dataset Summary
TitleHarnessing the power of electrophoresis and chromatography: Offline coupling of reverse phase liquid chromatography-capillary zone electrophoresis-tandem mass spectrometry for peptide mapping for monoclonal antibodies
DescriptionRPLC-MS/MS is the present workhorse for bottom-up proteomics-based characterization. However it suffers from limited peak capacity and challenges with respect to detection of small and hydrophilic peptides. CZE-MS/MS offers an orthogonal alternative to RPLC-MS/MS but has limited sensitivity due to low sample loading. In the present work, for the first time, offline coupling of an RPLC-CZE-MS/MS has been demonstrated for peptide mapping of a monoclonal antibody-based therapeutic product. The performance of this platform has been compared to the state-of-the-art RPLC-MS/MS and CZE-MS/MS approaches. Fifteen fractions were isolated from a mAb tryptic digest using RPLC, and each fraction was analyzed by CZE-ESI-MS/MS. Results indicate that not only the RPLC-CZE-MS/MS platform identify a larger number of distinct peptides (372) than the RPLC-MS/MS (219) and CZE-MS/MS (177) platforms, it also offered significantly superior sequence coverage (99.55% vs. 89.76% and 94.21% for the heavy chain and 98.6% vs. 92.06% and 95.79% for the light chain). Also, the distribution of amino acid residues with post translational modifications was 413 for RPLC-CZE-MS/MS, 150 for RPLC-MS/MS and 94 for CZE-MS/MS. In fact, the RPLC-CZE-MS/MS system performed better than the combined datasets from RPLC-MS/MS or CZE-MS/MS. Our study shows that the traditionally used one-dimensional (1D) RPLC-MS/MS and CZE-MS/MS-based platforms may not be enough for characterization of complex molecules such as monoclonal antibodies. The proposed approach should be an essential addition to the analytical toolkit for in-depth primary structural characterization of mAbs.
HostingRepositoryPRIDE
AnnounceDate2020-03-05
AnnouncementXMLSubmission_2020-03-05_08:23:12.xml
DigitalObjectIdentifier
ReviewLevelPeer-reviewed dataset
DatasetOriginOriginal dataset
RepositorySupportUnsupported dataset by repository
PrimarySubmitterAnurag Singh Rathore
SpeciesList scientific name: Mus musculus (Mouse); NCBI TaxID: 10090;
ModificationListmonohydroxylated residue; deamidated residue
InstrumentTripleTOF 5600
Dataset History
RevisionDatetimeStatusChangeLog Entry
02020-01-29 03:49:01ID requested
12020-03-05 08:23:14announced
Publication List
Kumar R, Shah RL, Rathore AS, Harnessing the power of electrophoresis and chromatography: Offline coupling of reverse phase liquid chromatography-capillary zone electrophoresis-tandem mass spectrometry for peptide mapping for monoclonal antibodies. J Chromatogr A, 1620():460954(2020) [pubmed]
Keyword List
ProteomeXchange project tag: Antibodies (B/D-HPP), Human Proteome Project
submitter keyword: Biotherapeutics, peptide mapping, RPLCMSMS,CZEMSMS
Contact List
Anurag Singh Rathore
contact affiliationProfessor of Chemical Enginnering Department Indian Institute of Technology Delhi
contact emailarathore@chemical.iitd.ac.in
lab head
Anurag Singh Rathore
contact affiliationProfessor Chemical Engineering Department. Indian Institute of Technology, Delhi.
contact emailarathore@chemical.iitd.ac.in
dataset submitter
Full Dataset Link List
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/2020/03/PXD017316
PRIDE project URI
Repository Record List
[ + ]