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PXD019497-1

PXD019497 is an original dataset announced via ProteomeXchange.

Dataset Summary
TitleImproved host cell protein analysis in monoclonal antibody products through ProteoMiner
DescriptionHost cell proteins (HCPs) impurities are critical quality attributes that have the potential to negatively impact the quality and safety profile of a biopharmaceutical product. Since HCPs may often arebe present at low levels, developing therefore highly a sensitive analytical method is needed to for their identification and quantitation is critical for process optimization and improvement to reduce them in the final drug product.ensure the safety and efficacy of the biopharmaceutical product. While an enzyme-linked immunosorbent assay (ELISA) can capture and quantify overall HCP levels, liquid chromatography coupled to mass spectrometry (LC-MS) is emerging as a powerful tool to monitor individual HCP levels during the purification process development. The massive dynamic range of protein species present in a therapeutic antibody is a major challenge for mass spectrometry-based methods to detect low-abundance HCP impurities. This study reports a powerful strategy to identify HCPs in an antibody drug substance by applying ProteoMiner enrichment followed by shotgun proteomic analysis. Using this strategy, we observed that the low abundance HCPs were enriched up to 1000-fold. In addition, by spiking in known amounts of HCPs to purified antibody drug substance with low levels of HCPs, we demonstrated that our method could detect HCP at a concentration as low as 0.05 ppm. When applying this methodology to the study of HCPs in NIST monoclonal antibody (NISTmAb), more than 500 HCPs were confidently identified, which tripled the number of identified HCPs that have been previously reported. Parallel reaction monitoring (PRM) results confirmed that the novel HCPs found using this method were enriched between 100 to 400-fold, highlighting that our method enriches and equalizes all proteins thus improving the sensitivity of HCP identification and quantification.
HostingRepositoryjPOST
AnnounceDate2021-06-02
AnnouncementXMLSubmission_2021-06-01_08:00:04.356.xml
DigitalObjectIdentifier
ReviewLevelNon peer-reviewed dataset
DatasetOriginOriginal dataset
RepositorySupportUnsupported dataset by repository
PrimarySubmitterI-Hsuan Chen
SpeciesList scientific name: Mus musculus (Mouse); NCBI TaxID: 10090;
ModificationListS-carboxamidomethyl-L-cysteine; L-methionine sulfoxide
InstrumentQ Exactive
Dataset History
RevisionDatetimeStatusChangeLog Entry
02020-06-01 12:08:57ID requested
12021-06-01 08:00:04announced
22022-09-18 03:56:34announced2022-09-18: Updated FTP location.
Publication List
Dataset with its publication pending
Keyword List
submitter keyword: Host cell Protein, ProteoMiner
Contact List
Hui Xiao
lab head
I-Hsuan Chen
contact affiliationRegeneron Pharmaceuticals
dataset submitter
Full Dataset Link List
jPOST dataset URI
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.biosciencedbc.jp/archive/jpostrepos/JPST000861