PXD035837

PXD035837 is an original dataset announced via ProteomeXchange.

Title	Fast & High Throughput LC-MS Characterization, and Peptide Mapping of Engineered AAV Capsids Using LC-MS/MS
Description	Adeno-associated virus (AAV) has emerged as a leading platform for gene therapy. With a skyrocketing rate of AAV research and the prevalence of many new engineered capsids being investigated in preclinical and clinical trials, capsid characterization plays an important role in serotype confirmation and quality control. Further, peptide mapping the capsid proteins might inevitably be a future requirement by regulatory agencies since it is a critical step in good manufacturing practice (GMP) for biotherapeutic characterization. To overcome many challenges that traditional methods like SDS-PAGE and Western blots carry, liquid chromatography & mass spectrometry (LC-MS) allows high resolution & sensitivity with great confidence in characterizing the AAV capsid proteins to the exact masses. Our optimized LC-MS method provides quick sample preparation, fast and high-throughput 4-min run, high sensitivity (only need ~2E10 vg per run for UV detection) allowing very efficient characterization of wild-type and engineered capsids. Additionally, , as well as LC-MS/MS peptide mapping of the AAV capsid proteins, including. lLysine-targeted chemical modification of the AAV capsids, for the first time, was characterized and peptide mapped in this study, therefore elucidating the most accessible Lysine residues of the AAV tested. Our detailed protocols method areis anticipated to promote the development and discovery of AAV variants with high accuracy and efficiency.
HostingRepository	PRIDE
AnnounceDate	2023-11-14
AnnouncementXML	Submission_2023-11-14_08:38:15.851.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Anh Lam
SpeciesList	scientific name: Adeno-associated virus 9; NCBI TaxID: 235455;
ModificationList	phosphorylated residue; acetylated residue; monohydroxylated residue; deamidated residue
Instrument	6545 Q-TOF LC/MS

Revision	Datetime	Status	ChangeLog Entry
0	2022-08-07 03:02:40	ID requested
1	2023-03-10 23:16:22	announced
⏵ 2	2023-11-14 08:38:16	announced	2023-11-14: Updated project metadata.

Lam AK, Zhang J, Frabutt D, Mulcrone PL, Li L, Zeng L, Herzog RW, Xiao W, Fast and high-throughput LC-MS characterization, and peptide mapping of engineered AAV capsids using LC-MS/MS. Mol Ther Methods Clin Dev, 27():185-194(2022) [pubmed]

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

submitter keyword: peptide map.,AAV capsids, engineered AAV capsids, LCMS, MSMS

Anh Kim Lam
contact affiliation	Indiana University
contact email	aklam@iu.edu
lab head
Anh Lam
contact affiliation	Indiana Univeristy School of Medicine
contact email	aklam@iu.edu
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD035837
     1. Label: PRIDE project
     2. Name: Fast & High Throughput LC-MS Characterization, and Peptide Mapping of Engineered AAV Capsids Using LC-MS/MS