, AG50-WX2 resin (Dowex, Dow Chemical Company

. Acetonitrile, , vol.99

H. Glycoclean and . Cartridges,

, 5-dihydroxybenzoic acid (Gentisic acid, vol.2

, Angiotensin I (1296.6853), Glu 1 -Fibrinopeptide B (1570.6774 Da), Da)

, TSK Amide-80 normal-phase HPLC column, 4.6 mm × 250 mm

, Acetonitrile 212, of low fluorescence grade for HPLC (Caledon

, 2AB) solution: 0.35 M 2AB solution in dimethyl sulfoxide/glacial acetic acid (7/3, v/v) containing 1 M sodium cyanoborohydride solution ( see Note 3 )

, 2AB-labeled dextran ladder

. 3mm and . Paper,

H. Glycoclean and . Cartridges,

, Solvent A: 50 mM ammonium formate

B. Solvent,

, GlcNAc 2 standard isolated from Phospholipase A 2 from bee venom as the most abundant N-glycan, and further characterized by, Man, vol.3

C. Saint-jore-dupas, L. Faye, and V. Gomord, From planta to pharma with glycosylation in the toolbox, Trends Biotechnol, vol.1, pp.317-323, 2007.

A. Fitchette-lainé, V. Gomord, M. Cabanes, J. Michalski, M. Saint-macary et al., N -glycans harboring the Lewis a epitope are expressed at the surface of plant cells, Plant J, vol.12, pp.1411-1417, 1997.

M. Bardor, G. Cabrera, P. M. Rudd, R. A. Dwek, J. A. Cremata et al., Analytical strategies to investigate plant N -glycan profiles in the context of plant-made pharmaceuticals, Curr. Opin. Struct. Biol, vol.16, pp.576-583, 2006.

L. Faye and M. J. Chrispeels, Characterization of N -linked oligosaccharides by affinoblotting with concanavalin A-peroxidase and treatment of the blots with glycosidases, Anal. Biochem, vol.149, pp.218-224, 1985.

N. Shibuya, I. J. Goldstein, E. J. Van-damme, and W. J. Peumans, Binding properties of a mannose-specific lectin from the snowdrop ( Galanthus nivealis ) bulb, J. Biol. Chem, vol.263, pp.728-734, 1988.

T. Animashaun, N. Mahmood, A. J. Hay, and R. C. Hughes, Inhibitory effect of novel mannose-binding lectins on HIVinfectivity and syncitium formation, Antiv. Chem. Chemother, vol.4, pp.145-153, 1993.

Y. Nagata and M. M. Burger, Wheat germ agglutinin. Molecular characteristics and specificity for sugar binding, J. Biol. Chem, vol.249, pp.3116-3122, 1974.

L. Faye, V. Gomord, A. Fitchette-lainé, and M. J. Chrispeels, Affinity purification of antibodies specific for Asn-linked glycans containing ?1-> 3 fucose or ?1-> 2 xylose, Anal. Biochem, vol.209, pp.104-108, 1993.

N. Tomiya, M. Kurono, H. Ishihara, S. Tejima, S. Endo et al., Structural analysis of N -linked oligosaccharides by a combination of glycopeptidase, exoglycosidases, and highperformance liquid chromatography, Anal. Biochem, vol.163, pp.489-499, 1987.

H. Ogawa, A. Hijikata, M. Amano, H. Fukushima, I. Ishizuka et al., Structures and contribution to the antigenicity of oligosaccharides of Japanese cedar ( Cryptomeria japonica) pollen allergen Cry j I: relationship between the structures and antigenic epitopes of plant N -linked complex-type glycans, Glycoconj. J, vol.13, pp.555-566, 1996.

M. Séveno, G. Cabrera, A. Trigero, C. Burel, J. Leprince et al., Plant N-glycan profiling of minute amounts of material, Anal. Biochem, vol.379, issue.1, pp.66-72, 2008.

H. Bakker, M. Bardor, J. W. Molthoff, V. Gomord, I. Elbers et al., Galactose-extended glycans of antibodies produced by transgenic plants, Proc. Natl. Acad. Sci. USA, vol.98, issue.5, pp.2899-2904, 2001.

C. Rayon, M. Cabanes-macheteau, C. Loutelier-bourrhis, I. Saliot-maire, J. Lemoine et al., Characterization of N -glycans from Arabidopsis thaliana . Application to a fucose-deficient mutant, Plant Physiol, vol.119, pp.725-733, 1999.

M. Cabanes-macheteau, A. Fitchette-lainé, C. Loutelier-bourhis, C. Lange, N. D. Vine et al., N -glycosylation of a mouse IgG expressed in transgenic tobacco plants, Glycobiology, vol.9, pp.365-372, 1999.

M. Bardor, M. Cabanes-macheteau, L. Faye, and P. Lerouge, Monitoring the N -glycosylation by fluorophore-assisted carbohydrate electrophoresis, vol.21, pp.2550-2556, 2000.

K. Ko, Y. Tekoah, P. M. Rudd, D. J. Harvey, R. A. Dwek et al., Function and glycosylation of plant-derived antiviral monoclonal antibody, Proc. Natl. Acad. Sci. USA, vol.100, pp.8013-8018, 2003.

A. Triguero, G. Cabrera, J. A. Cremata, C. Yuen, J. Wheeler et al., Plant-derived mouse IgG monoclonal antibody fused to KDEL endoplasmic reticulum-retention signal is N -glycosylated homogeneously throughout the plant with mostly high-mannose-type N -glycans, Plant Biotech. J, vol.3, pp.449-457, 2005.
DOI : 10.1111/j.1467-7652.2005.00137.x

URL : https://onlinelibrary.wiley.com/doi/pdf/10.1111/j.1467-7652.2005.00137.x

G. R. Guile, P. M. Rudd, D. R. Wing, S. B. Prime, and R. A. Dwek, A rapid high-resolution high-performance liquid chromatographic method for separating glycan mixtures and analyzing oligosaccharide profiles, Anal. Biochem, vol.240, pp.210-226, 1996.
DOI : 10.1006/abio.1996.0351

P. M. Rudd, C. Colominas, L. Royle, N. Murphy, E. Hart et al., A high-performance liquid chromatography based strategy for rapid, sensitive sequencing of N -linked oligosaccharide modifications to proteins in sodium dodecyl sulphate polyacrylamide electrophoresis gel bands, Proteomics, vol.1, pp.285-294, 2001.

D. J. Harvey, R. A. Dwek, and P. M. Rudd, Determining the structure of glycan moieties by mass spectrometry, Current Protocols in Protein Science, 2006.

L. Royle, C. M. Radcliffe, R. A. Dwek, and P. M. Rudd, Detailed structural analysis of N -glycans released from glycoproteins in SDS-PAGE gel bands using HPLC combined with exoglycosidase array digestions, Methods in Molecular Biology, vol.347, pp.125-144, 2006.

M. Bardor, C. Loutelier-bourhis, and L. Marvin, Analysis of plant glycoproteins by matrix-assisted laser desorption ionisation mass spectrometry: application to the N -glycosylation of bean phytohemagglutinin, Plant Physiol. Biochem, vol.37, pp.319-325, 1999.

M. Bardor, L. Faye, and P. Lerouge, Analysis of the N -glycosylation of recombinant glycoproteins produced in transgenic plants, Trends Plant Sci, vol.4, pp.376-380, 1999.

C. R. Bird, A. J. Gearing, and R. Thorpe, The use of Tween-20 alone as a blocking agent for the immunoblotting can cause artefactual results, J. Immunol. Methods, vol.106, pp.175-179, 1988.

A. Lainé and L. Faye, Significant immunological cross-reactivity of plant glycoproteins, Electrophoresis, vol.9, pp.841-844, 1988.

V. Kubelka, F. Altmann, E. Staudacher, V. Trotter, L. März et al., Primary structures of the N -linked carbohydrate chains from honeybee venom phospholipase A2, Eur. J. Biochem, vol.213, pp.1193-1204, 1993.

A. Vitale, T. G. Warner, and M. J. Chrispeels, Phaseolus vulgaris phytohemagglutinin contains high-mannose and modified oligoasaccharide chains, Planta, vol.160, pp.256-263, 1984.

M. Bardor, C. Loutelier-bourhis, T. Paccalet, P. Cosette, A. Fitchette et al., Monoclonal C5-1 antibody produced in transgenic alfalfa plants exhibits a N -glycosylation that is homogeneous and suitable for glyco-engineering into a human-compatible structure, Plant Biotech. J, vol.1, pp.451-462, 2003.