HUMAN POLYREACTIVE AND MONOREACTIVE ANTIBODIES - EFFECT OF GLYCOSYLATION ON ANTIGEN-BINDING
HUMAN POLYREACTIVE AND MONOREACTIVE ANTIBODIES - EFFECT OF GLYCOSYLATION ON ANTIGEN-BINDING
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DOI：
10.1093/glycob/4.4.491
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发表时间：
1994-08-01
期刊：
影响因子：
4.3
通讯作者：
HARINDRANATH, N
中科院分区：
文献类型：
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作者：
DONADEL, G;CALABRO, A;HARINDRANATH, N
启动本实验以确定抗体分子的碳水化合物部分是否有助于多反应性。用衣霉素处理产生免疫球蛋白（Ig）M、IgG和伊加同种型的人单克隆多反应性或单反应性抗体的细胞系，以阻断蛋白质的N-连接糖基化。对分泌的天然和非糖基化蛋白的分析揭示了对[H-3]甘露糖掺入的> 95%抑制。十二烷基硫酸钠-聚丙烯酰胺凝胶电泳的蛋白质从衣霉素处理的细胞显示增加的流动性和[H-3]甘露糖掺入的免疫球蛋白重链的情况下，与缺乏糖基化。然后测试天然和非糖基化抗体结合不同抗原的能力。尽管缺乏糖基化，但多反应性和单反应性抗体都与抗原结合，反应性或特异性几乎没有损失。它的结论是，碳水化合物部分没有显着贡献的多反应性。
The present experiments were initiated to determine whether the carbohydrate portions of antibody molecules contribute to polyreactivity. Cell lines making human monoclonal polyreactive or monoreactive antibodies of the imnunoglobulin (lg) M, IgG and IgA isotypes were treated with tunicamycin to block N-linked glycosylation of the proteins. Analysis of the secreted native and non-glycosylated proteins revealed a > 95% inhibition of [H-3]mannose incorporation. Electrophoresis on sodium dodecyl sulphate-polyacrylamide gels of the proteins from tunicamycin-treated cells showed increased mobility and the absence of [H-3]mannose incorporation of the immunoglobulin heavy chains, consistent with the lack of glycosylation. The native and non-glycosylated antibodies were then tested for their ability to bind different antigens. Despite the lack of glycosylation, both polyreactive and monoreactive antibodies bound to antigens with little if any loss of reactivity or specificity. It is concluded that the carbohydrate moieties do not contribute significantly to polyreactivity.