在植物中生产基于设计的 VHH 抗体。

Production of Designer VHH-Based Antibodies in Plants.

机构信息

VIB-VUB Center for Structural Biology, Brussels, Belgium.

Structural Biology Brussels, Vrije Universiteit Brussel, Brussels, Belgium.

出版信息

Methods Mol Biol. 2022;2446:205-230. doi: 10.1007/978-1-0716-2075-5_10.

DOI:10.1007/978-1-0716-2075-5_10

PMID:35157275

Abstract

Simplified monoclonal antibodies can be produced by fusing a VHH or nanobody, derived from camelid heavy-chain-only antibodies to the Fc domain of either IgG (VHH-IgG), IgA (VHH-IgA), or IgY (VHH-IgY). These recombinant antibodies are encoded by a single gene and their production can be easily scaled up in plants. This chapter contains methods for Gateway cloning of VHH-Fc fusions into the binary T-DNA vectors pEAQ-HT-DEST1 and pPhasGW, electroporation of Agrobacterium with the resulting constructs, transient antibody expression in Nicotiana benthamiana leaves, and stable antibody expression in Arabidopsis thaliana seeds. The properties of chimeric VHH-based antibodies produced in plants enable novel passive immunization treatments, such as in-feed oral delivery or intravenous injection.

摘要

通过将源自骆驼重链抗体的 VHH 或纳米抗体与 IgG（VHH-IgG）、IgA（VHH-IgA）或 IgY（VHH-IgY）的 Fc 结构域融合，可以生产简化的单克隆抗体。这些重组抗体由单个基因编码，其生产可以在植物中轻松放大。本章包含了将 VHH-Fc 融合物通过 Gateway 克隆到二元 T-DNA 载体 pEAQ-HT-DEST1 和 pPhasGW 中的方法、用构建物电穿孔农杆菌、在 Nicotiana benthamiana 叶片中瞬时表达抗体以及在拟南芥种子中稳定表达抗体。在植物中产生的基于 VHH 的嵌合抗体的特性使新型被动免疫治疗成为可能，例如经饲料口服递送或静脉内注射。