Boes Alexander, Reimann Andreas, Twyman Richard M, Fischer Rainer, Schillberg Stefan, Spiegel Holger
Fraunhofer Institute for Molecular Biology and Applied Ecology IME, Forckenbeckstrasse 6, 52074, Aachen, Germany.
TRM Ltd, PO Box 93, York, YO43 3WE, UK.
Methods Mol Biol. 2016;1404:597-619. doi: 10.1007/978-1-4939-3389-1_39.
There are currently no vaccines that provide sterile immunity against malaria. Various proteins from different stages of the Plasmodium falciparum life cycle have been evaluated as vaccine candidates, but none of them have fulfilled expectations. Therefore, combinations of key antigens from different stages of the parasites life cycle may be essential for the development of efficacious malaria vaccines. Following the identification of promising antigens using bioinformatics, proteomics, and/or immunological approaches, it is necessary to express, purify, and characterize these proteins and explore the potential of fusion constructs combining different antigens or antigen domains before committing to expensive and time-consuming clinical development. Here, using malaria vaccine candidates as an example, we describe how Agrobacterium tumefaciens-based transient expression in plants can be combined with a modular and flexible cloning strategy as a robust and versatile tool for the rapid production of candidate antigens during research and development.
目前尚无能够提供针对疟疾的无菌免疫的疫苗。恶性疟原虫生命周期不同阶段的各种蛋白质已被评估为候选疫苗,但它们均未达到预期效果。因此,来自寄生虫生命周期不同阶段的关键抗原组合对于开发有效的疟疾疫苗可能至关重要。在使用生物信息学、蛋白质组学和/或免疫学方法鉴定出有前景的抗原之后,有必要对这些蛋白质进行表达、纯化和表征,并在投入昂贵且耗时的临床开发之前探索结合不同抗原或抗原结构域的融合构建体的潜力。在此,我们以疟疾候选疫苗为例,描述了基于根癌农杆菌的植物瞬时表达如何与模块化且灵活的克隆策略相结合,作为一种强大且通用的工具,用于在研发过程中快速生产候选抗原。