Department of Biological & Agricultural Engineering, Texas A&M University, College Station, TX 77843, United States.
Biotechnol Adv. 2012 Mar-Apr;30(2):419-33. doi: 10.1016/j.biotechadv.2011.07.020. Epub 2011 Aug 6.
Plants are becoming commercially acceptable for recombinant protein production for human therapeutics, vaccine antigens, industrial enzymes, and nutraceuticals. Recently, significant advances in expression, protein glycosylation, and gene-to-product development time have been achieved. Safety and regulatory concerns for open-field production systems have also been addressed by using contained systems to grow transgenic plants. However, using contained systems eliminates several advantages of open-field production, such as inexpensive upstream production and scale-up costs. Upstream technological achievements have not been matched by downstream processing advancements. In the past 10 years, the most research progress was achieved in the areas of extraction and pretreatment. Extraction conditions have been optimized for numerous proteins on a case-by-case basis leading to the development of platform-dependent approaches. Pretreatment advances were made after realizing that plant extracts and homogenates have unique compositions that require distinct conditioning prior to purification. However, scientists have relied on purification methods developed for other protein production hosts with modest investments in developing novel plant purification tools. Recently, non-chromatographic purification methods, such as aqueous two-phase partitioning and membrane filtration, have been evaluated as low-cost purification alternatives to packed-bed adsorption. This paper reviews seed, leafy, and bioreactor-based platforms, highlights strategies for the primary recovery and purification of recombinant proteins, and compares process economics between systems. Lastly, the future direction and research needs for developing economically competitive recombinant proteins with commercial potential are discussed.
植物作为用于生产人类治疗药物、疫苗抗原、工业用酶和营养保健品的重组蛋白的商业生产方法正逐渐被接受。目前，在表达、蛋白质糖基化和基因到产物的开发时间方面已取得了重大进展。通过使用封闭系统来种植转基因植物，已经解决了露天生产系统的安全和监管问题。但是，使用封闭系统消除了露天生产的几个优势，例如廉价的上游生产和规模化成本。上游技术成就并没有与下游加工的进步相匹配。在过去的 10 年中，在提取和预处理领域取得了最多的研究进展。针对许多蛋白质，已经根据具体情况优化了提取条件，从而开发了基于平台的方法。在意识到植物提取物和匀浆具有独特的组成成分，需要在纯化之前进行独特的调理后，预处理方面取得了进展。然而，科学家们一直依赖于为其他蛋白质生产宿主开发的纯化方法，而在开发新型植物纯化工具方面的投资很少。最近，非色谱纯化方法（如双水相萃取和膜过滤）已被评估为填充床吸附的低成本替代方法。本文综述了种子、叶状和生物反应器平台，重点介绍了用于初级回收和重组蛋白纯化的策略，并比较了不同系统之间的工艺经济性。最后，讨论了开发具有商业潜力的经济竞争力的重组蛋白的未来方向和研究需求。
