PXD018229-2

PXD018229 is an original dataset announced via ProteomeXchange.

Title	Analysis of coagulation factor IX in bioreactor cell culture media predicts yield and quality of purified product
Description	Coagulation factor IX (FIX) is a highly complex post-translationally modified human serum glycoprotein and a high-value biopharmaceutical. The quality of recombinant FIX (rFIX), especially complete -carboxylation, is critical for rFIX clinical efficacy. Changes in bioreactor operating conditions during rFIX production are likely to have a strong effect on both protein production and the occupancy and structure of rFIX post-translational modifications (PTMs). We hypothesized that monitoring the bioreactor cell culture supernatant with Data Independent Acquisition Mass Spectrometry (MS) would allow is to predict of product yield and quality after purification. With the ultimate goal of optimizing rFIX production, we developed a suite of MS proteomics analytical methods and used these to investigate changes in rFIX yield, -carboxylation, and other PTMs, as well as host cell proteins during bioreactor culture and after purification. Our methods provided a detailed overview of the dynamics of site-specific PTM occupancy and abundance on rFIX during production, which accurately predicted the efficiency of purification and the quality of the purified product from different culture conditions. In addition, we identified new PTMs in rFIX near the GLA domain, which could impact rFIX GLA-dependent purification efficiency and protein function. The workflows presented here are applicable to other biologics and expression systems, and should aid in the optimization and quality control of upstream and downstream bioprocesses.
HostingRepository	PRIDE
AnnounceDate	2024-10-22
AnnouncementXML	Submission_2024-10-22_05:20:28.084.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Lucia Zacchi
SpeciesList	scientific name: Cricetulus griseus (Chinese hamster) (Cricetulus barabensis griseus); NCBI TaxID: 10029;
ModificationList	S-carboxamidoethyl-L-cysteine
Instrument	TripleTOF 5600

Revision	Datetime	Status	ChangeLog Entry
0	2020-03-27 02:50:17	ID requested
1	2021-03-28 17:14:22	announced
⏵ 2	2024-10-22 05:20:28	announced	2024-10-22: Updated project metadata.

Dataset with its publication pending

submitter keyword: gamma-carboxylation, mass spectrometry, glycoproteomics, CHO, host cell proteins,coagulation factor

Benjamin Schulz
contact affiliation	School of Chemistry and Molecular Biosciences, The University of Queensland
contact email	b.schulz@uq.edu.au
lab head
Lucia Zacchi
contact affiliation	The University of Queensland
contact email	l.zacchi@uq.edu.au
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD018229
     1. Label: PRIDE project
     2. Name: Analysis of coagulation factor IX in bioreactor cell culture media predicts yield and quality of purified product