Stable expression of chimeric heavy chain antibodies in CHO cells.
Stable expression of chimeric heavy chain antibodies in CHO cells.
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DOI：
10.1007/978-1-61779-968-6_18
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发表时间：
2012-01-01
期刊：
影响因子：
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通讯作者：
Durocher, Yves
中科院分区：
文献类型：
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作者：
Agrawal, Vishal;Slivac, Igor;Durocher, Yves
与非骆驼科动物Fc区融合的骆驼科单域抗体，也称为嵌合重链抗体（cHCAb），由于其相对较小的尺寸（80 kDa）和完整的Fc，作为治疗和诊断候选物具有巨大潜力。在本章中，我们描述了两种方法，即有限稀释法和小池法，用于在悬浮培养和无血清培养中使用严格的抗生素筛选产生稳定表达cHCAb的非扩增中国仓鼠卵巢细胞系。这两种方案都不需要购置或使用昂贵的自动化设备，因此可以在任何具有最基本细胞培养设施的实验室中应用。所提供的方案能够分离出在摇瓶中进行分批培养时能够产生高达100 mg/L抗体的稳定克隆。
Camelid single domain antibodies fused to noncamelid Fc regions, also called chimeric heavy chain antibodies (cHCAb), offer great potential as therapeutic and diagnostic candidates due to their relatively small size (80 kDa) and intact Fc. In this chapter, we describe two approaches, limiting dilution and minipools, for generating nonamplified Chinese hamster ovary cell lines stably expressing cHCAb in suspension and serum-free cultures using a stringent antibiotic selection. Neither of the protocols necessitates the acquisition or implementation of expensive automated infrastructures and thus could be applied in any lab with minimal cell culture setup. The given protocol allows the isolation of stable clones capable of generating up to 100 mg/L of antibody in batch mode performed in shaker flasks.