Antibody production using a ciliate generates unusual antibody glycoforms displaying enhanced cell-killing activity.
Antibody production using a ciliate generates unusual antibody glycoforms displaying enhanced cell-killing activity.
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DOI：
10.1080/19420862.2016.1228504
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发表时间：
2016
期刊：
影响因子：
5.3
通讯作者：
Crispin M
中科院分区：
文献类型：
--
作者：
Calow J;Behrens AJ;Mader S;Bockau U;Struwe WB;Harvey DJ;Cormann KU;Nowaczyk MM;Loser K;Schinor D;Hartmann MW;Crispin M
抗体糖基化是优化抗体治疗的关键参数。在这里，我们描述了单细胞纤毛虫，嗜热四膜虫抗癌单克隆抗体利妥昔单抗的生产。由此产生的抗体表现出增强的抗体依赖细胞介导的细胞毒性，我们将其归因于不寻常的n -链糖基化。详细的色谱和质谱分析揭示了一个集中的，寡甘露聚糖型的聚糖，作为一个整体，显示出与典型的人类对应的异构体结构偏离，但其分支相当于在人类糖蛋白中观察到的代谢中间产物的片段。从沉积的晶体结构分析，我们预测纤毛虫聚糖与Fc结构域的蛋白质-碳水化合物相互作用与天然络合型聚糖非常相似。此外，还鉴定了与人类糖基化的生物合成前体相匹配的末端葡萄糖结构。我们的研究结果表明，基于纤毛虫的表达系统提供了一种大规模生产单克隆抗体的途径，这种单克隆抗体表现出糖基化，从而增强了细胞杀伤活性。
Antibody glycosylation is a key parameter in the optimization of antibody therapeutics. Here, we describe the production of the anti-cancer monoclonal antibody rituximab in the unicellular ciliate, Tetrahymena thermophila. The resulting antibody demonstrated enhanced antibody-dependent cell-mediated cytotoxicity, which we attribute to unusual N-linked glycosylation. Detailed chromatographic and mass spectrometric analysis revealed afucosylated, oligomannose-type glycans, which, as a whole, displayed isomeric structures that deviate from the typical human counterparts, but whose branches were equivalent to fragments of metabolic intermediates observed in human glycoproteins. From the analysis of deposited crystal structures, we predict that the ciliate glycans adopt protein-carbohydrate interactions with the Fc domain that closely mimic those of native complex-type glycans. In addition, terminal glucose structures were identified that match biosynthetic precursors of human glycosylation. Our results suggest that ciliate-based expression systems offer a route to large-scale production of monoclonal antibodies exhibiting glycosylation that imparts enhanced cell killing activity.
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影响因子：
7.3
作者：
Frenzel A;Hust M;Schirrmann T
通讯作者：
Schirrmann T
影响因子：
3.7
作者：
Cowan GJ;Bockau U;Eleni-Muus J;Aldag I;Samuel K;Creasey AM;Hartmann MW;Cavanagh DR
通讯作者：
Cavanagh DR
影响因子：
15
作者：
Bowden, Thomas A.;Baruah, Kavitha;Coles, Charlotte H.;Harvey, David J.;Yu, Xiaojie;Song, Byeong-Doo;Stuart, David I.;Aricescu, A. Radu;Scanlan, Christopher N.;Jones, E. Yvonne;Crispin, Max
通讯作者：
Crispin, Max
影响因子：
4.3
作者：
Goetze, Andrew M.;Liu, Y. Diana;Flynn, Gregory C.
通讯作者：
Flynn, Gregory C.
影响因子：
5.6
作者：
Crispin, Max;Bowden, Thomas A.;Jones, E. Yvonne
通讯作者：
Jones, E. Yvonne