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PXD053035

PXD053035 is an original dataset announced via ProteomeXchange.

Dataset Summary
TitleDevelopment of an optimized LC-MS workflow for Host Cell Protein characterization to support upstream process development
DescriptionHost cell proteins (HCPs) co-expressed during the production of biotherapeutics can affect the safety, efficacy and stability of the final product. As such, monitoring HCPs populations and amounts throughout the production and purification process are an essential part of the overall quality control framework. Mass spectrometry (MS)is used as an orthogonal method to enzyme-linked immunosorbent assays (ELISA) for simultaneous identification and quantification of HCPs, particularly for the analysis of downstream processes. In this study, we present an MS-based analytical protocol with improvements in both speed and identification performance that can be implemented for routine analysis to support upstream process development. The protocol adopts a streamlined sample preparation strategy combined with a high-throughput MS analysis pipeline. The developed method identifies and quantifies over 1000 HCPs, including 20 proteins listed as high risk in literature, in a clarified cell culture sample with repeatability and precision shown for digest replicates. In addition, we explore the effects of varying standard spike-ins and changes to the data processing pipeline on absolute quantification estimates of the HCPs, which highlights the importance of standardization for the wider use in industry.
HostingRepositoryPRIDE
AnnounceDate2025-05-07
AnnouncementXMLSubmission_2025-05-06_19:20:20.118.xml
DigitalObjectIdentifier
ReviewLevelPeer-reviewed dataset
DatasetOriginOriginal dataset
RepositorySupportUnsupported dataset by repository
PrimarySubmitterJanik Seidel
SpeciesList scientific name: Cricetulus griseus (Chinese hamster) (Cricetulus barabensis griseus); NCBI TaxID: 10029;
ModificationListacetylated residue; monohydroxylated residue; iodoacetamide derivatized residue
InstrumentZenoTOF 7600
Dataset History
RevisionDatetimeStatusChangeLog Entry
02024-06-12 04:24:49ID requested
12025-05-06 19:20:21announced
Publication List
10.1021/acs.jproteome.4c00637;
Seidel JD, Condina MR, Klingler-Hoffmann M, Young C, Donnellan L, Kyngdon C, Hoffmann P, Development of an Optimized LC-MS Workflow for Host Cell Protein Characterization to Support Upstream Process Development. J Proteome Res, 24(1):234-243(2025) [pubmed]
Keyword List
submitter keyword: process analytical technologies,Host cell proteins, LC-MS, clarified cell culture fluid, absolute quantification, Chinese hamster ovary, data-independent acquisition, hi3 quantification, bio-processing
Contact List
Prof. Peter Hoffmann
contact affiliationClinical and Health Sciences, Mass Spectrometry and Proteomics facility, University of South Australia, Adelaide, Australia
contact emailPeter.Hoffmann@unisa.edu.au
lab head
Janik Seidel
contact affiliationUniversity of South Australia (UniSA)
contact emailseijy006@mymail.unisa.edu.au
dataset submitter
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Dataset FTP location
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