High-level expression of recombinant IgG in the human cell line PER.C6
High-level expression of recombinant IgG in the human cell line PER.C6
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DOI：
10.1021/bp025574h
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发表时间：
2003-01-01
影响因子：
2.9
通讯作者：
Bout, A
中科院分区：
文献类型：
--
作者：
Jones, D;Kroos, N;Bout, A
预计生产中的治疗性单克隆抗体数量将在未来几年迅速增加。因此，抗体表达平台的优化备受关注。有几个问题很重要，包括从实验室过渡到生产的速度、IgG 的产量和质量（特别是免疫球蛋白上存在的聚糖结构）。我们已经表征了人类细胞系 PER.C6 的生产重组 IgG 的能力。产量仍在优化中，但在非补料分批培养中，PER.C6 能够生长至 5x10(6) 个细胞/mL 的细胞密度，并产生 300-500 mg/L IgG；在补料分批培养中，这种情况可能会显着增加。产生抗体的细胞系的产生速度很快，因为高产量不需要插入基因的多轮扩增。插入基因的基因拷贝数在每个基因组1-10个拷贝的范围内。此外，PER.C6 是人类细胞系，因此不添加对人类具有免疫原性的聚糖。核心岩藻糖分子基本上始终存在，半乳糖残基以生理水平存在（每个聚糖有 0、1 和 2 个半乳糖残基，比例为 1:2:1）。没有看到杂合或高甘露糖结构。
The number of therapeutic monoclonal antibodies in production is expected to rise rapidly in the next few years. As a result, there is much focus on the optimization of antibody expression platforms. Several issues are important including the speed of transition from bench to manufacturing, yield of IgG, and quality (particularly of the glycan structures present on immunoglobulins). We have characterized the human cell line PER.C6 for its ability to produce recombinant IgG. Production yields are still being optimized, but in nonfed batch culture, PER.C6 is able to grow to a cell density of 5x10(6) cells/mL and produce 300-500 mg/L IgG; this is likely to increase significantly in fed batch cultures. The generation of antibody-producing cell lines is fast, as rounds of amplification of inserted genes are not required for high production yields. The gene copy number of inserted genes is in the region of 1-10 copies per genome. In addition, PER.C6 is a human cell line, and so does not add glycans, which are immunogenic in humans. A core fucose molecule is essentially always present, and galactose residues are present at a physiological level (0, 1, and 2 galactose residues per glycan are present at a ratio of 1:2:1). No hybrid or high-mannose structures are seen.