Journal: Glycobiology
Article Title: N-glycomic profiling of capsid proteins from Adeno-Associated Virus serotypes
doi: 10.1093/glycob/cwad074
Figure Lengend Snippet: Representative exoglycosidases sequential digestion array for InstantPC labeled N-glycans from AAV6 serotype by HILIC-FLD. The initial N-glycan profile prior to digestion is displayed in . N-glycan profiles subjected to specific exoglycosidase enzyme arrays are shown as follows: A) Αlpha (1-2,3,6)-mannosidase (Jack bean, JBM) to cleave terminal, non-reducing α (1-2,3,6)-linked mannose. B) Sialidase A ( Arthrobacter ureafaciens , ABS) to cleave linear and branched terminal, non-reducing α2-3,6,8,9-linked sialic acid. C) β (1-3,4)-galactosidase (bovine testis, BTG) to cleave terminal β (1-3,4)-linked galactose. D) β-(1-2,3,4,6)-hexosaminidase (jack bean, GUH) to cleave terminal, non-reducing β-(1-2,3,4,6)-linked N-acetylglucosamine (GlcNAc) and N-acetylgalactosamine (GalNAc). Higher enzyme concentrations may be necessary to remove bisecting β-(1-4)-linked GlcNAc. E) α (1-2,4,6)-Fucosidase O (Omnitrophica bacterium, FUCO) to cleave α (1-2,4,6)-linked fucose, but it cleaves α (1-6) core fucose more efficiently than other linkages. The numbers shown above the peaks refer to the structures in the initial dataset in the —AAV N-glycan assignments). The new N-glycan peaks after exoglycosidase sequential digestion are labeled as sequential digestion followed by numbers (SD1-72) and included in the secondary dataset in —Exoglycosidase sequential digestion). The dataset contains 81 N-glycan structures identified in the analysis prior to digestion (1-81) and 72 new N-glycan structures identified as exoglycosidases sequential digestion (SD1-72).
Article Snippet: The AdvanceBio Gly-X N-Glycan Prep with InstantPC kit, 96-ct (Part NO: GX96-IPC) consisting of three modules, including Gly-X deglycosylation module (Part NO: GX96-100), Gly-X InstantPC labeling module (Part NO: GX96-101), and Gly-X InstantPC Clean-up module (Part NO: GX96-102), AdvanceBio exoglycosidases of α (1-2,3,6)-Mannosidase (jack bean, Part NO: GKX-5010, JBM), Sialidase A ( Arthrobacter ureafaciens , Part NO: GK80040, ABS), β (1-3,4)-Galactosidase (bovine testis, Part NO: GKX5013, BTG), β-N-acetylhexosaminidase (jack bean, Part NO: GK80050, GUH), and InfinityLab Ultrapure LC/MS acetonitrile (Part NO: 5191-4496) were donated by Agilent Technologies (Santa Clara, CA 95051, U.S.A.). α (1-2,4,6)-Fucosidase O (Omnitrophica bacterium, Part NO: P0749, FUCO) were purchased from New England Biolabs (Ipswich, MA 01938-2723, USA).
Techniques: Labeling, Hydrophilic Interaction Liquid Chromatography, Glycoproteomics
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