RT Journal Article SR Electronic T1 High Throughput Isolation and Glycosylation Analysis of IgG–Variability and Heritability of the IgG Glycome in Three Isolated Human Populations JF Molecular & Cellular Proteomics JO Mol Cell Proteomics FD American Society for Biochemistry and Molecular Biology SP M111.010090 DO 10.1074/mcp.M111.010090 VO 10 IS 10 A1 Pučić, Maja A1 Knežević, Ana A1 Vidič, Jana A1 Adamczyk, Barbara A1 Novokmet, Mislav A1 Polašek, Ozren A1 Gornik, Olga A1 Šupraha-Goreta, Sandra A1 Wormald, Mark R. A1 Redžić, Irma A1 Campbell, Harry A1 Wright, Alan A1 Hastie, Nicholas D. A1 Wilson, James F. A1 Rudan, Igor A1 Wuhrer, Manfred A1 Rudd, Pauline M. A1 Josić, Djuro A1 Lauc, Gordan YR 2011 UL http://www.mcponline.org/content/10/10/M111.010090.abstract AB All immunoglobulin G molecules carry N-glycans, which modulate their biological activity. Changes in N-glycosylation of IgG associate with various diseases and affect the activity of therapeutic antibodies and intravenous immunoglobulins. We have developed a novel 96-well protein G monolithic plate and used it to rapidly isolate IgG from plasma of 2298 individuals from three isolated human populations. N-glycans were released by PNGase F, labeled with 2-aminobenzamide and analyzed by hydrophilic interaction chromatography with fluorescence detection. The majority of the structural features of the IgG glycome were consistent with previous studies, but sialylation was somewhat higher than reported previously. Sialylation was particularly prominent in core fucosylated glycans containing two galactose residues and bisecting GlcNAc where median sialylation level was nearly 80%. Very high variability between individuals was observed, approximately three times higher than in the total plasma glycome. For example, neutral IgG glycans without core fucose varied between 1.3 and 19%, a difference that significantly affects the effector functions of natural antibodies, predisposing or protecting individuals from particular diseases. Heritability of IgG glycans was generally between 30 and 50%. The individual's age was associated with a significant decrease in galactose and increase of bisecting GlcNAc, whereas other functional elements of IgG glycosylation did not change much with age. Gender was not an important predictor for any IgG glycan. An important observation is that competition between glycosyltransferases, which occurs in vitro, did not appear to be relevant in vivo, indicating that the final glycan structures are not a simple result of competing enzymatic activities, but a carefully regulated outcome designed to meet the prevailing physiological needs.