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Comparison of separation techniques for the elucidation of IgG N-glycans pooled from healthy mammalian species

Artikel i vetenskaplig tidskrift
Författare Barbara Adamczyk
Tharmala Tharmalingam-Jaikarana
Michael Schomberg
Ákos Szekrényes
Ronan M. Kelly
Niclas G. Karlsson
Andràs Guttman
Pauline M. Rudd
Publicerad i Carbohydrate Research
Volym 389
Sidor 174-185
ISSN 0008-6215
Publiceringsår 2014
Publicerad vid Institutionen för biomedicin, avdelningen för medicinsk kemi och cellbiologi
Sidor 174-185
Språk en
Länkar dx.doi.org/10.1016/j.carres.2014.01...
Ämnesord Protein glycosylation; IgG; Glycan analysis; HILIC–UPLC; RP–UPLC; CE-LIF
Ämneskategorier Biomedicinsk laboratorievetenskap/teknologi

Sammanfattning

The IgG N-glycome provides sufficient complexity and information content to serve as an excellent source for biomarker discovery in mammalian health. Since oligosaccharides play a significant role in many biological processes it is very important to understand their structure. The glycosylation is cell type specific as well as highly variable depending on the species producing the IgG. We evaluated the variation of N-linked glycosylation of human, bovine, ovine, equine, canine and feline IgG using three orthogonal glycan separation techniques: hydrophilic interaction liquid chromatography (HILIC)–UPLC, reversed phase (RP)–UPLC and capillary electrophoresis with laser induced fluorescence detection (CE-LIF). The separation of the glycans by these high resolution methods yielded different profiles due to diverse chemistries. However, the % abundance of structures obtained by CE-LIF and HILIC–UPLC were similar, whereas the analysis by RP-UPLC was difficult to compare as the structures were separated by classes of glycans (highly mannosylated, fucosylated, bisected, fucosylated and bisected) resulting in the co-elution of many structures. The IgGs from various species were selected due to the complexity and variation in their N-glycan composition thereby highlighting the complementarity of these separation techniques.

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