Quantitation of the oligosaccharides of human serum IgG from patients with rheumatoid arthritis: a critical evaluation of different methods (original) (raw)
1998, Journal of Immunological Methods
Several different chromatographic methods and a lectin-based assay have been compared for the quantitation of Ž . oligosaccharides released from immunoglobulin G IgG . The analysis of a series of IgG samples purified from the serum of rheumatoid arthritis patients was carried out by these methods to evaluate the percentage of the glycoforms having 0, 1 or 2 Ž . Ž. Ž. galactose residues G0, G1 and G2 in order to a identify the method that can be most widely used for quantitation, b Ž . accurately define the range of G0 values found in patients with rheumatoid arthritis, and c make available a series of characterised standards for distribution to clinical chemistry laboratories. The chromatographic methods involved: release of oligosaccharides by glycoamidase A after protease digestion followed by HPLC analysis of aminopyridine derivatives on Ž . reverse phase and normal phase columns; hydrazinolysis treatment with exoglycosidases G0 mix and Biogel P4 Ž . chromatography of 2-aminobenzamide 2-AB derivatives; hydrazinolysis and weak anion exchange or normal phase HPLC of 2-AB derivatives; release of oligosaccharides by PNGase F and either Biogel P4 chromatography of 2-AB derivatives or HPAEC-PAD analysis of native oligosaccharides. The G0 values given by these methods compared favourably with each other and a dot blot assay of denatured IgG interaction with Ricinus communis agglutinin and Bandeiraea simplicifolia S 0 0 2 2 -1 7 5 9 9 8 0 0 0 3 2 -5 ( ) F. H. Routier et al.r Journal of Immunological Methods 213 1998 113-130 114 lectin II. The HPLC and HPAEC methods give additional information that may be important in less routine assays. q 1998 Elsevier Science B.V. All rights reserved.
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