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在植物中生产骆驼状抗体。

Production of camel-like antibodies in plants.

作者信息

De Buck Sylvie, Virdi Vikram, De Meyer Thomas, De Wilde Kirsten, Piron Robin, Nolf Jonah, Van Lerberge Els, De Paepe Annelies, Depicker Ann

机构信息

Department of Plant Systems Biology, VIB, Ghent, Belgium.

出版信息

Methods Mol Biol. 2012;911:305-24. doi: 10.1007/978-1-61779-968-6_19.

Abstract

Transgenic plants for the production of high-value recombinant complex and/or glycosylated proteins are a promising alternative for conventional systems, such as mammalian cells and bacteria. Many groups use plants as production platform for antibodies and antibody fragments. Here, we describe how bivalent camel-like antibodies can be produced in leaves and seeds. Camel-like antibodies are fusions of the antigen-binding domain of heavy chain camel antibodies (VHH) with an Fc fragment of choice. Transient expression in Nicotiana benthamiana leaves allows the production of VHH-Fc antibodies within a few days after the expression plasmid has been obtained. Generation of stable Arabidopsis thaliana transformants allows production of scalable amounts of VHH-Fc antibodies in seeds within a year. Further, we describe how the in planta-produced VHH-Fc antibodies can be quantified by Western blot analysis with Fc-specific antibodies.

摘要

用于生产高价值重组复合物和/或糖基化蛋白的转基因植物是传统系统(如哺乳动物细胞和细菌)的一种有前途的替代方案。许多研究小组将植物用作抗体和抗体片段的生产平台。在此,我们描述了如何在叶片和种子中生产双价骆驼源抗体。骆驼源抗体是重链骆驼抗体(VHH)的抗原结合结构域与所选Fc片段的融合体。在获得表达质粒后几天内,通过在本氏烟草叶片中瞬时表达可产生VHH-Fc抗体。生成稳定的拟南芥转化体可在一年内从种子中生产出可扩展数量的VHH-Fc抗体。此外,我们还描述了如何通过使用Fc特异性抗体的蛋白质免疫印迹分析对植物中产生的VHH-Fc抗体进行定量。

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