PXD029305

PXD029305 is an original dataset announced via ProteomeXchange.

Title	Host Cell Protein Quantification Workflow Using Optimized Standards combined with Data-Independent Acquisition Mass Spectrometry
Description	Monitoring of host cell proteins (HCPs) during the manufacture of monoclonal antibodies (mAb) has become a critical requirement to provide effective and safe drug product. ELISA assays are still the current gold standard for the quantification of protein impurities. However, this technique has several limitations and does, among others, not enable the precise identification of proteins. In this context, mass spectrometry (MS) has emerged as an alternative and orthogonal method that delivers qualitative and quantitative information on all identified HCPs. However, in order to be routinely implemented in biopharmaceutical companies, liquid chromatography (LC)-MS based methods still need to be standardized to provide highest sensitivity and robust and accurate quantification. In this study, we developed a promising MS-based analytical workflow coupling the use of an innovative quantification standard, the HCP Profiler solution, with a spectral library-based data-independent acquisition (DIA) method and strict data validation criteria. The performance of the HCP Profiler solution was compared to more conventional standard protein spikes and the DIA approach was benchmarked against classical data-dependent acquisition (DDA) using samples collected at various stages of the manufacturing process. Finally, we further explored the possibility to use spectral library-free DIA. As a result, our method showed accurate and reproducible (coefficients of variation (CVs) < 10%) quantification of HCPs while reaching a sensitivity down to the sub-ng/mg mAb level.
HostingRepository	PRIDE
AnnounceDate	2023-11-14
AnnouncementXML	Submission_2023-11-14_09:03:46.641.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Steve Hessmann
SpeciesList	scientific name: Cricetulus griseus (Chinese hamster) (Cricetulus barabensis griseus); NCBI TaxID: 10029;
ModificationList	acetylated residue; monohydroxylated residue; iodoacetamide derivatized residue
Instrument	Q Exactive HF-X

Revision	Datetime	Status	ChangeLog Entry
0	2021-10-24 23:57:24	ID requested
1	2023-07-20 12:09:46	announced
⏵ 2	2023-11-14 09:03:50	announced	2023-11-14: Updated project metadata.

Hessmann S, Chery C, Sikora AS, Gervais A, Carapito C, Host cell protein quantification workflow using optimized standards combined with data-independent acquisition mass spectrometry. J Pharm Anal, 13(5):494-502(2023) [pubmed]

submitter keyword: HCP Profiler, protein quantification, data validation., data-independent acquisition (DIA),host cell proteins (HCPs)

Christine Carapito
contact affiliation	Laboratoire de Spectrométrie de Masse BioOrganique (LSMBO), Université de Strasbourg, CNRS, IPHC, UMR 7178, 25 Rue Becquerel, F-67087 Strasbourg, France
contact email	ccarapito@unistra.fr
lab head
Steve Hessmann
contact affiliation	LSMBO IPHC CNRS
contact email	steve.hessmann2@etu.unistra.fr
dataset submitter

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/2023/07/PXD029305

PRIDE project URI

• PRIDE
  1. PXD029305
     1. Label: PRIDE project
     2. Name: Host Cell Protein Quantification Workflow Using Optimized Standards combined with Data-Independent Acquisition Mass Spectrometry