PXD042062-1

PXD042062 is an original dataset announced via ProteomeXchange.

Title	Variation of Site-Specific Glycosylation Profiles of Recombinant Influenza Glycoproteins
Description	Recombinant proteins are of great interest in glycobiology and proteomics, known especially for their reproducibility and accessibility. However, variation in glycosylation among recombinant glycoproteins is not well understood and may depend on numerous conditions in the biomanufacturing process. In order to confidently assess variation in glycosylation measurements, it is vital to both optimize the measurement of, and determine the degree of variation between, distributions of glycosylation on specific sites of glycoproteins. This is especially important for glycoproteins that are known to have rapid sequence changes, such as with different influenza strains. In this study, eight strains of recombinant influenza hemagglutinin and neuraminidase produced from HEK293 cell line were obtained from four vendors and digestion was conducted using a series of complex multi-enzymatic methods designed to isolate glycopeptide sequons. Site-specific glycosylation profiles of intact glycopeptides were produced using mass spectrometric evaluation on an orbitrap system and compared using spectral similarity scores. Variation in glycan abundances and distribution was most pronounced between different strains of virus (similarity score = 383 out of 1000), whereas replicates resulted in low variation (similarity score = 957 out of 1000). Glycan variation was also measured based on differences between vendors, lots, batches, protease digestion, and intra-protein site. The most abundant glycans in all of these influenza glycoproteins were monofucosylated and complex, as reported by other laboratories. However, it was found that different vendors can produce very different glycan distributions for the same glycosylation site. Notably, it is demonstrated that glycan distributions are similar for conserved regions of influenza glycoproteins. Overall, these methods present a potential use in developing reproducible measurements of glycosylated biologics for quality control or making more informed decisions in biomanufacturing.
HostingRepository	PRIDE
AnnounceDate	2024-08-13
AnnouncementXML	Submission_2024-08-13_04:39:19.901.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Zachary Goecker
SpeciesList	scientific name: Influenza A virus (A/Hong Kong/485197/2014(H3N2)); NCBI TaxID: 1679038; scientific name: Influenza A virus (A/Netherlands/219/2003(H7N7)); NCBI TaxID: 680693; scientific name: Influenza A virus (A/New Caledonia/20/1999(H3N2)); NCBI TaxID: 480020; scientific name: Influenza A virus (A/Hong Kong/483/1997(H5N1)); NCBI TaxID: 88104; scientific name: Influenza A virus (A/Thailand/1(KAN-1)/2004(H5N1)); NCBI TaxID: 266827; scientific name: Influenza A virus (A/Japan/305/1957(H2N2)); NCBI TaxID: 387161; scientific name: Influenza A virus (A/Arizona/13/2008(H1N1)); NCBI TaxID: 593933; scientific name: Influenza A virus (A/California/04/2009(H1N1)); NCBI TaxID: 641501;
ModificationList	iodoacetamide derivatized residue
Instrument	Orbitrap Fusion Lumos

Revision	Datetime	Status	ChangeLog Entry
0	2023-05-08 12:02:24	ID requested
⏵ 1	2024-08-13 04:39:20	announced

10.1016/J.MCPRO.2024.100827;

submitter keyword: Glycosylation,Influenza A, Site-specific, Hemagglutinin

Stephen E.
contact affiliation	Biomolecular Measurement Division, National Institute of Standards and Technology
contact email	stephen.stein@nist.gov
lab head
Zachary Goecker
contact affiliation	National Institute of Standards and Technology
contact email	zachary.goecker@nist.gov
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD042062
     1. Label: PRIDE project
     2. Name: Variation of Site-Specific Glycosylation Profiles of Recombinant Influenza Glycoproteins