PXD053035 is an
original dataset announced via ProteomeXchange.
Dataset Summary
Title | Development of an optimized LC-MS workflow for Host Cell Protein characterization to support upstream process development |
Description | Host 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. |
HostingRepository | PRIDE |
AnnounceDate | 2025-05-07 |
AnnouncementXML | Submission_2025-05-06_19:20:20.118.xml |
DigitalObjectIdentifier | |
ReviewLevel | Peer-reviewed dataset |
DatasetOrigin | Original dataset |
RepositorySupport | Unsupported dataset by repository |
PrimarySubmitter | Janik Seidel |
SpeciesList | scientific name: Cricetulus griseus (Chinese hamster) (Cricetulus barabensis griseus); NCBI TaxID: 10029; |
ModificationList | acetylated residue; monohydroxylated residue; iodoacetamide derivatized residue |
Instrument | ZenoTOF 7600 |
Dataset History
Revision | Datetime | Status | ChangeLog Entry |
0 | 2024-06-12 04:24:49 | ID requested | |
⏵ 1 | 2025-05-06 19:20:21 | announced | |
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 affiliation | Clinical and Health Sciences, Mass Spectrometry and Proteomics facility, University of South Australia, Adelaide, Australia |
contact email | Peter.Hoffmann@unisa.edu.au |
lab head | |
Janik Seidel |
contact affiliation | University of South Australia (UniSA) |
contact email | seijy006@mymail.unisa.edu.au |
dataset submitter | |
Full Dataset Link List
Dataset FTP location
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PRIDE project URI |
Repository Record List
[ + ]
[ - ]
- PRIDE
- PXD053035
- Label: PRIDE project
- Name: Development of an optimized LC-MS workflow for Host Cell Protein characterization to support upstream process development