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Open AccessArticle

Site-Specific N-Glycosylation on the AAV8 Capsid Protein

1
Center for Diagnostics & Therapeutics and Department of Chemistry, Georgia State University, Atlanta, GA 30302, USA
2
Sol Sherry Thrombosis Research Center, Lewis Katz, School of Medicine, Temple University, Philadelphia, PA 19140, USA
3
School of Biomedical Science, Huaqiao University, Quanzhou 362021, China
*
Authors to whom correspondence should be addressed.
These two authors contribute equally to the study.
Viruses 2018, 10(11), 644; https://doi.org/10.3390/v10110644
Received: 2 October 2018 / Revised: 5 November 2018 / Accepted: 15 November 2018 / Published: 17 November 2018
Adeno associated virus (AAV) is a versatile gene delivery tool, which has been approved as a human gene therapy vector for combating genetic diseases. AAV capsid proteins are the major components that determine the tissue specificity, immunogenicity and in vivo transduction performance of the vector. In this study, the AAV8 capsid glycosylation profile was systemically analyzed by peptide mass fingerprinting utilizing high-resolution mass spectrometry to determine the presence of capsid glycosylation. We identified N-glycosylation on the amino acid N499 of the capsid protein. We characterized the overall sugar profile for vector produced in 293 cells. Multiple N-glycosylated host-cell proteins (HCPs) copurified with AAV8 vectors and were identified by analyzing LC-MS data utilizing a human database and proteome discoverer search engine. The N-glycosylation analysis by MALDI-TOF MS, highlighted the probability of AAV8 interaction with terminal galactosylated N-glycans within the HCPs. View Full-Text
Keywords: Adeno associated virus; mass spectrometry; site specific N-glycan analysis; host cell protein analysis; virus-host interaction Adeno associated virus; mass spectrometry; site specific N-glycan analysis; host cell protein analysis; virus-host interaction
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Aloor, A.; Zhang, J.; Gashash, E.A.; Parameswaran, A.; Chrzanowski, M.; Ma, C.; Diao, Y.; Wang, P.G.; Xiao, W. Site-Specific N-Glycosylation on the AAV8 Capsid Protein. Viruses 2018, 10, 644.

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