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PXD017214

PXD017214 is an original dataset announced via ProteomeXchange.

Dataset Summary
TitleS-Trap eliminates Pluronic F68 and cell culture media polymers for effective proteomic analysis of mammalian cell bioreactor supernatants
DescriptionProteomic analysis of bioreactor supernatants can inform on cellular metabolic status, viability, and productivity, as well as product quality, which can in turn help optimize bioreactor operation. Incubating mammalian cells in bioreactors requires the addition of polymeric surfactants such as Pluronic F68, which reduce the sheer stress caused by agitation. However, these surfactants are incompatible with mass spectrometry proteomics and must be eliminated during sample preparation. Here, we compared four different sample preparation methods to eliminate Pluronic F68 from filtered bioreactor supernatant samples: organic solvent precipitation; filter-assisted sample preparation (FASP); S-Trap; and single-pot, solid-phase, sample preparation (SP3). We found that SP3 and S-Trap substantially reduced or eliminated the polymer(s), but S-Trap provided the most robust clean-up and highest quality data. Additionally, we observed that SP3 sample preparation of our samples and in other published datasets was associated with partial alkylation of cysteines, which could impact the confidence and robustness of protein identification and quantification. Finally, we observed that several commercial mammalian cell culture media and media supplements also contained polymers with a similar mass spectrometry profile to Pluronic F68, and we suggest that proteomic analyses in these media will also benefit from the use of S-Trap sample preparation.
HostingRepositoryPRIDE
AnnounceDate2020-06-30
AnnouncementXMLSubmission_2020-06-30_00:13:28.xml
DigitalObjectIdentifier
ReviewLevelPeer-reviewed dataset
DatasetOriginOriginal dataset
RepositorySupportUnsupported dataset by repository
PrimarySubmitterLucia Zacchi
SpeciesList scientific name: Homo sapiens (Human); NCBI TaxID: 9606; scientific name: Saccharomyces cerevisiae (Baker's yeast); NCBI TaxID: 4932;
ModificationListNo PTMs are included in the dataset
InstrumentTripleTOF 5600
Dataset History
RevisionDatetimeStatusChangeLog Entry
02020-01-20 08:22:48ID requested
12020-06-30 00:13:29announced
Publication List
Zacchi LF, Recinos DR, Otte E, Aitken C, Hunt T, Sandford V, Lee YY, Schulz BL, Howard CB, S-Trap Eliminates Cell Culture Media Polymeric Surfactants for Effective Proteomic Analysis of Mammalian Cell Bioreactor Supernatants. J Proteome Res, 19(5):2149-2158(2020) [pubmed]
Keyword List
submitter keyword: mass spectrometry, polymer contaminants, alkylation, sample preparation, bottom-up proteomics, bioreactor samples, S-trap, SP3
Contact List
Benjamin Schulz
contact affiliationSchool of Chemistry and Molecular Biosciences, The University of Queensland
contact emailb.schulz@uq.edu.au
lab head
Lucia Zacchi
contact affiliationThe University of Queensland
contact emaill.zacchi@uq.edu.au
dataset submitter
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