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PXD072803

PXD072803 is an original dataset announced via ProteomeXchange.

Dataset Summary
TitleLC-MS peptide mapping of monoclonal antibodies using the mirror proteases trypsin and Tryp-N
DescriptionAccurate characterization of the amino acid sequence and post-translational modifications (PTMs) of monoclonal antibodies (mAbs) is essential for evaluating product quality. Peptide mapping through bottom-up LC/MS analysis is a key methodology for this purpose. While trypsin is commonly the first choice for mAb digestion, it typically yields high but incomplete sequence coverage. As a result, supplementary endoproteases such as Asp-N, chymotrypsin, Glu-C or Lys-C are often employed to enhance coverage. In this report, we evaluated another endoprotease, Tryp-N, which serves as an effective alternative to trypsin for mAb analysis. The sequence coverages achieved for bevacizumab, cetuximab, NISTmAb, and trastuzumab with Tryp-N were comparable to that of trypsin, and the combination of both enzymes slightly improved overall sequence coverage. Notably, both trypsin and Tryp-N generated identical peptides beside the N- and C-terminal ends. The presence of a basic amino acid at opposite ends of the peptide often resulted in complementary sequence coverage of the MS2 of the same peptide sequences. These complementary ion series can be leveraged for precise localization of PTMs, as demonstrated in detail for deamidation, and oxidation sites as well as single amino acid variations (SAVs).
HostingRepositoryjPOST
AnnounceDate2026-04-15
AnnouncementXMLSubmission_2026-04-14_08:00:04.838.xml
DigitalObjectIdentifier
ReviewLevelPeer-reviewed dataset
DatasetOriginOriginal dataset
RepositorySupportUnsupported dataset by repository
PrimarySubmitterBernd Thiede
SpeciesList scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationListS-carboxamidomethyl-L-cysteine; L-methionine sulfoxide; unknown modification
InstrumenttimsTOF Pro
Dataset History
RevisionDatetimeStatusChangeLog Entry
02026-01-08 00:39:09ID requested
12026-04-14 08:00:06announced
Publication List
Sargautis D, Thiede B, LC-MS peptide mapping of monoclonal antibodies using the mirror proteases trypsin and Tryp-N. J Pharm Biomed Anal, 272():117361(2026) [pubmed]
Keyword List
submitter keyword: Monoclonal antibody, mirror protease, NISTmAb, peptide mapping, posttranslational modification, single variant amino acid, Tryp-N
Contact List
Bernd Thiede
lab head
Bernd Thiede
contact affiliationUniversity of Oslo
dataset submitter
Full Dataset Link List
jPOST dataset URI
Dataset FTP location
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