High throughput isolation and glycosylation analysis of IgG-variability and heritability of the IgG glycome in three isolated human populations.
High throughput isolation and glycosylation analysis of IgG-variability and heritability of the IgG glycome in three isolated human populations.
复制标题
DOI：
10.1074/mcp.m111.010090
复制
发表时间：
2011-10
期刊：
影响因子：
--
通讯作者：
Lauc G
中科院分区：
文献类型：
--
作者：
Pucić M;Knezević A;Vidic J;Adamczyk B;Novokmet M;Polasek O;Gornik O;Supraha-Goreta S;Wormald MR;Redzić I;Campbell H;Wright A;Hastie ND;Wilson JF;Rudan I;Wuhrer M;Rudd PM;Josić D;Lauc G
所有免疫球蛋白G分子都携带N-聚糖，N-聚糖调节其生物活性。IgG的N-糖基化的变化与各种疾病相关，并影响治疗性抗体和静脉内免疫球蛋白的活性。我们已经开发了一种新的96孔蛋白G整体板，并使用它来快速分离IgG从血浆中的2298个人从三个孤立的人群。通过PNGase F释放N-聚糖，用2-氨基苯甲酰胺标记，并通过亲水相互作用色谱法结合荧光检测进行分析。IgG糖组的大多数结构特征与先前的研究一致，但唾液酸化略高于先前报道的。唾液酸化在含有两个半乳糖残基和二等分GlcNAc的核心岩藻糖基化聚糖中尤其突出，其中中位唾液酸化水平接近80%。观察到个体之间的变异性非常高，约为总血浆糖组的3倍。例如，不含核心岩藻糖的中性IgG聚糖在1.3%至19%之间变化，这一差异显著影响天然抗体的效应子功能，从而诱发或保护个体免受特定疾病的影响。IgG聚糖的遗传率通常在30%至50%之间。个体的年龄与半乳糖的显著减少和二等分GlcNAc的增加相关，而IgG糖基化的其他功能元件并不随年龄变化。性别不是任何IgG聚糖的重要预测因素。一个重要的观察结果是，在体外发生的糖基转移酶之间的竞争似乎在体内并不相关，这表明最终的聚糖结构不是竞争酶活性的简单结果，而是一个精心调节的结果，旨在满足普遍的生理需求。
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.