Human IgG preparations isolated by ion-exchange or protein G affinity chromatography differ in their glycosylation profiles.
IgG from patients with rheumatoid arthritis (RA) is abnormally glycosylated in the Fc region, with sialic acid and galactose levels lower than normal. Protein G and DEAE purify populations which are differentially glycosylated. Significantly increased exposure of sialic acid was detected in normal IgG compared with that of RA IgG when ion exchange was used to prepare samples. However, when the same samples were prepared using protein G, no difference in the detection of sialic acid was seen between the two groups. When examining the heavy chain of IgG, more sialic acid, galactose and N-acetylglucosamine were detected in DEAE purified IgG compared with that prepared by protein G Detection of sialic acid and N-acetylglucosamine was also increased on light chains from IgG prepared by ion exchange chromatography. Since this occurs notably on rheumatoid light chains it would appear that this arrangement would contribute to the overall glycosylation changes in IgG. In the case of molecules lacking galactose the discrimination between the RA and normal IgG is significantly improved when ion exchange chromatography is used. Since differentiation between disease and normal groups relies on the purification technique used, we recommend that more than one method is employed before undertaking an analysis of glycosylation changes.
Bond
A
A
Department of Cellular and Molecular Sciences, St. George's Hospital Medical School, London, UK.
Jones
M G
MG
Hay
F C
FC
D001172:Arthritis, Rheumatoid / Q000276:immunology; D002241:Carbohydrates / Q000737:chemistry; D002846:Chromatography, Affinity; D002848:Chromatography, DEAE-Cellulose; D006031:Glycosylation; D006801:Humans; D015151:Immunoblotting; D007074:Immunoglobulin G / Q000737:chemistry* / Q000302:isolation & purification*; D007143:Immunoglobulin Heavy Chains / Q000737:chemistry / Q000302:isolation & purification; D007147:Immunoglobulin Light Chains / Q000737:chemistry / Q000302:isolation & purification; D037102:Lectins; D009419:Nerve Tissue Proteins; D037121:Plant Lectins; D006023:Glycoproteins
D002241:Carbohydrates; D007074:Immunoglobulin G; D007143:Immunoglobulin Heavy Chains; D007147:Immunoglobulin Light Chains; D037102:Lectins; D009419:Nerve Tissue Proteins; D037121:Plant Lectins; C029152:G-substrate; C017707:Ricinus communis agglutinin-1; C046179:glycosylated IgG; D006023:Glycoproteins