Chen Qiang, Lai Huafang, Hurtado Jonathan, Stahnke Jake, Leuzinger Kahlin, Dent Matthew
The Biodesign Institute, Center for Infectious Disease and Vaccinology, Arizona State University, USA.
College of Technology and Innovation, Arizona State University, USA.
Adv Tech Biol Med. 2013 Jun;1(1). doi: 10.4172/atbm.1000103.
Current human biologics are most commonly produced by mammalian cell culture-based fermentation technologies. However, its limited scalability and high cost prevent this platform from meeting the ever increasing global demand. Plants offer a novel alternative system for the production of pharmaceutical proteins that is more scalable, cost-effective, and safer than current expression paradigms. The recent development of deconstructed virus-based vectors has allowed rapid and high-level transient expression of recombinant proteins, and in turn, provided a preferred plant based production platform. One of the remaining challenges for the commercial application of this platform was the lack of a scalable technology to deliver the transgene into plant cells. Therefore, this review focuses on the development of an effective and scalable technology for gene delivery in plants. Direct and indirect gene delivery strategies for plant cells are first presented, and the two major gene delivery technologies based on agroinfiltration are subsequently discussed. Furthermore, the advantages of syringe and vacuum infiltration as gene delivery methodologies are extensively discussed, in context of their applications and scalability for commercial production of human pharmaceutical proteins in plants. The important steps and critical parameters for the successful implementation of these strategies are also detailed in the review. Overall, agroinfiltration based on syringe and vacuum infiltration provides an efficient, robust and scalable gene-delivery technology for the transient expression of recombinant proteins in plants. The development of this technology will greatly facilitate the realization of plant transient expression systems as a premier platform for commercial production of pharmaceutical proteins.
目前的人用生物制品大多通过基于哺乳动物细胞培养的发酵技术生产。然而，其有限的可扩展性和高成本使得该平台无法满足全球不断增长的需求。植物为药用蛋白的生产提供了一种新型替代系统，该系统比目前的表达模式更具可扩展性、成本效益更高且更安全。基于解构病毒的载体的最新发展使得重组蛋白能够快速、高水平地瞬时表达，进而提供了一个首选的植物生产平台。该平台商业应用的一个遗留挑战是缺乏一种可扩展的技术将转基因导入植物细胞。因此，本综述重点关注植物基因传递有效且可扩展技术的发展。首先介绍了植物细胞的直接和间接基因传递策略，随后讨论了基于农杆菌浸润的两种主要基因传递技术。此外，还广泛讨论了注射器浸润和真空浸润作为基因传递方法的优势，包括它们在植物中商业生产人用药物蛋白的应用和可扩展性。综述中还详细介绍了成功实施这些策略的重要步骤和关键参数。总体而言，基于注射器浸润和真空浸润的农杆菌浸润为植物中重组蛋白的瞬时表达提供了一种高效、稳健且可扩展的基因传递技术。该技术的发展将极大地促进植物瞬时表达系统成为药用蛋白商业生产的首要平台的实现。
