PXD016215-1

PXD016215 is an original dataset announced via ProteomeXchange.

Title	Profiling of a high mannose-type N-glycosylated lipase using hydrophilic interaction chromatography-mass spectrometry
Description	We report the use of hydrophilic interaction liquid chromatography-mass spectrometry (HILIC-MS) to profile intact glycoform distributions of high mannose-type N-glycosylated proteins, using an industrially produced fungal lipase for the food industry as an example. We compared these results with conventional reversed phase LC-MS (RPLC-MS) and sodium dodecyl sulfate–polyacrylamide gel-electrophoresis (SDS-PAGE). HILIC appeared superior in resolving lipase heterogeneity, facilitating mass assignment of N-glycoforms and sequence variants. Fractions representing the four main HILIC elution bands for lipase were subjected to SDS-PAGE confirming that HILIC retention increased with the number of glycosylation sites (0-3) occupied. Additional bottom-up proteomic analysis of the fractions enabled the identification of the most abundant glycosylation sites. Compared to RPLC-MS, HILIC-MS provided a stronger reduction of the sample complexity delivered to the mass spectrometer, facilitating the assignment of the masses of glycoforms and sequence variants as well as increasing the number of glycoforms detected (69 more proteoforms, 177% increase). The HILIC-MS method required relatively short analysis time (< 30 min), in which over 100 glycoforms were distinguished. We suggest that HILIC-MS can be used to characterize bioengineering processes aimed at steering protein glycoform expression as well as to check the consistency of product batches.
HostingRepository	PRIDE
AnnounceDate	2020-05-04
AnnouncementXML	Submission_2020-05-04_01:24:31.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Andrea Gargano
SpeciesList	scientific name: Aspergillus niger ATCC 13496; NCBI TaxID: 1353008;
ModificationList	complex glycosylation
Instrument	Q Exactive

Revision	Datetime	Status	ChangeLog Entry
0	2019-11-11 02:36:02	ID requested
⏵ 1	2020-05-04 01:24:32	announced

Dataset with its publication pending

ProteomeXchange project tag: Biology/Disease-Driven Human Proteome Project (B/D-HPP), Human Proteome Project, Glycoproteomics (B/D-HPP)

submitter keyword: High mannose-type N-glycosylation, intact protein analysis, HILIC, high-resolution mass spectrometry

Andrea Gargano
contact affiliation	University of Amsterdam
contact email	a.gargano@uva.nl
lab head
Andrea Gargano
contact affiliation	University of Amsterdam
contact email	a.gargano@uva.nl
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD016215
     1. Label: PRIDE project
     2. Name: Profiling of a high mannose-type N-glycosylated lipase using hydrophilic interaction chromatography-mass spectrometry