Genmab BV, AD Utrecht, the Netherlands.
Biotechnol Bioeng. 2010 Feb 1;105(2):350-7. doi: 10.1002/bit.22535.
Rapid production of recombinant human IgG with improved antibody dependent cell-mediated cytotoxicity (ADCC) effector function is presented. The technique employs transient expression of IgG in suspension growing HEK-293F cells in the presence of the glycosidase inhibitor kifunensine. The procedure takes approximately 7 days, provided that expression plasmids encoding the IgG of interest are available. Kifunensine inhibits the N-linked glycosylation pathway of HEK-293F cells in the endoplasmatic reticulum, resulting in IgG with oligomannose type glycans lacking core-fucose. IgG1 transiently produced in kifunensine- treated HEK-293F cells has improved affinity for the FcgammaRIIIA molecule as measured in an ELISA based assay, and almost eightfold enhanced ADCC using primary peripheral blood mononuclear effector cells.
本文提出了一种快速生产具有改善的抗体依赖细胞介导的细胞毒性（ADCC）效应功能的重组人 IgG 的方法。该技术采用在存在糖苷酶抑制剂 kifunensine 的情况下，在悬浮生长的 HEK-293F 细胞中瞬时表达 IgG。只要有编码所需 IgG 的表达质粒可用，该过程大约需要 7 天。Kifunensine 在 ER 中抑制 HEK-293F 细胞的 N 连接糖基化途径，导致 IgG 具有缺少核心岩藻糖的寡甘露糖型聚糖。用基于 ELISA 的测定法测量，在 kifunensine 处理的 HEK-293F 细胞中瞬时产生的 IgG1 对 FcγRIIIA 分子的亲和力提高，并且使用原代外周血单核细胞效应细胞的 ADCC 增强近 8 倍。
