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001-es BibID:BIBFORM099698
035-os BibID:(cikkazonosító)462593
Első szerző:Liénard--Mayor, Théo
Cím:High sensitivity capillary electrophoresis with fluorescent detection for glycan mapping / Liénard--Mayor Théo, Yang Bin, Tran Nguyet Thuy, Bruneel Arnaud, Guttman Andras, Taverna Myriam, Mai Thanh Duc
Dátum:2021
ISSN:0021-9673
Megjegyzések:We present in this study a novel strategy to drastically improve the detection sensitivity and peak capac- ity for capillary electrophoresis with laser induced fluorescent detection (CE-LIF) of glucose oligomers and released glycans. This is based on a new approach exploiting a polymer-free background electrolyte (BGE) for CE-LIF of glycans. The best performance in terms of sample stacking and suppression of electroos- motic flow (EOF) was found for a BGE composed of triethanolamine/citric acid and triethanolamine/acetic acid at elevated ionic strengths (IS up to 200 mM). Compared to the conventional protocols for CE- LIF of glucose-oligosaccharides and released glycans, our polymer-free strategy offered up to 5-fold im- provement of detection sensitivity and visualization of higher degree of polymerization (DP) of glucose oligomers (18 vs 15). To further improve the detection sensitivity, a new electrokinetic preconcentration strategy via large volume sample stacking with electroosmotic modulation without having recourse to neutrally coated capillaries is proposed, offering a 200-fold signal enhancement. This approach is based on variation of the buffer's IS, rather than pH adjustment as in conventional methods, for EOF modula- tion or quasi-total reduction. This strategy allows selecting with high flexibility the best pH conditions to perform efficient preconcentration and separation. The new approach was demonstrated to be applicable for the analysis of N-linked oligosaccharides released from a model glycoprotein (Human Immunoglobu- lin G) and applied to map N-glycans from human serum for congenital disorders of glycosylation (CDG) diagnosis.
Tárgyszavak:Orvostudományok Elméleti orvostudományok idegen nyelvű folyóiratközlemény külföldi lapban
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Megjelenés:Journal of Chromatography A. - 1657 (2021), p. 1-9. -
További szerzők:Yang, Bin Tran, N. Thuy Bruneel, Arnaud Guttman András (1954-) (vegyészmérnök) Taverna, Myriam Mai, Thanh Duc
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