PXD025971

PXD025971 is an original dataset announced via ProteomeXchange.

Title	Rapid Highly-Efficient Digestion and Peptide Mapping of Adeno-Associated Viruses
Description	[15:18] Toole, Estee Adeno-Associated Viruses (AAVs) comprise an area of rapidly growing interest due to their ability to act as a gene delivery vehicle in novel gene therapy strategies and vaccine development. Peptide mapping is a common technique in the biopharmaceutical industry to confirm the correct sequence, product purity, PTMs, and stability. However, conventional peptide mapping is time-consuming and has proven difficult to reproduce with viral capsids because of their high structural stability and the suboptimal localization of trypsin cleavage sites in the AAV protein sequences. In this study, we present an optimized workflow that provides thorough characterization within one day. This workflow is also highly reproducible due to its simplicity having very few steps, and easy to perform proteolytic digestion utilizing thermally-stable pepsin, active at 70°C in acidic conditions. The acidic conditions of the peptic digestions drive viral capsid denaturation and improve cleavage site accessibility. We characterized the efficiency and ease of digestion through peptide mapping of the AAV2 viral capsid protein. Using nanoflow liquid chromatography coupled with tandem mass spectrometry, we achieved 100% sequence coverage of the low abundance VP1 capsid protein with a digestion process taking only 10 minutes to prepare and 30 minutes to complete the digestion.
HostingRepository	PRIDE
AnnounceDate	2022-02-17
AnnouncementXML	Submission_2022-02-17_03:28:08.254.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Andrew Williamson
SpeciesList	scientific name: Adeno-associated virus - 2; NCBI TaxID: 10804;
ModificationList	monohydroxylated residue; deamidated residue
Instrument	Q Exactive

Revision	Datetime	Status	ChangeLog Entry
0	2021-05-12 01:46:22	ID requested
⏵ 1	2022-02-17 03:28:09	announced

submitter keyword: [15:18] Toole, Estee Gene Therapy, AAV, Viral Therapies, Peptide Mapping, Pepsin, Trypsin, nanoLC, proteolysis

Estee Toole
contact affiliation	Thermo Fisher Scientific
contact email	estee.toole@thermofisher.com
lab head
Andrew Williamson
contact affiliation	Thermo Fisher Scientific
contact email	andrew.williamson@thermofisher.com
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD025971
     1. Label: PRIDE project
     2. Name: Rapid Highly-Efficient Digestion and Peptide Mapping of Adeno-Associated Viruses