Site-Specific N-Glycosylation on the AAV8 Capsid Protein

被引：33
作者：
Aloor, Arya [1 ,2 ]
Zhang, Junping [3 ,4 ]
Gashash, Ebtesam A. [1 ,2 ]
Parameswaran, Aishwarya [1 ,2 ]
Chrzanowski, Matthew [3 ]
Ma, Cheng [1 ,2 ]
Diao, Yong [4 ]
Wang, Peng George [1 ,2 ]
Xiao, Weidong [3 ]
机构：
[1] Georgia State Univ, Ctr Diagnost & Therapeut, Atlanta, GA 30302 USA
[2] Georgia State Univ, Dept Chem, Atlanta, GA 30302 USA
[3] Temple Univ, Sch Med, Lewis Katz, Sol Sherry Thrombosis Res Ctr, Philadelphia, PA 19140 USA
[4] Huaqiao Univ, Sch Biomed Sci, Quanzhou 362021, Peoples R China
来源：
VIRUSES-BASEL | 2018年 / 10卷 / 11期
基金：
中国国家自然科学基金;
关键词：
Adeno associated virus; mass spectrometry; site specific N-glycan analysis; host cell protein analysis; virus-host interaction; ADENOASSOCIATED VIRUS SEROTYPE-8; SIALIC-ACID BINDING; GENE-THERAPY; VIRAL CAPSIDS; IDENTIFICATION; VECTORS; TRANSDUCTION; RECEPTOR; TYPE-2; EFFICIENCY;
D O I：
10.3390/v10110644
中图分类号：
Q93 [微生物学];
学科分类号：
071005 ; 100705 ;
摘要：
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.
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页数：14
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