Abstract
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.
Highlights
Plants have been explored as alternatives to mammalian, insect, and bacterial cell cultures for the expression and production of recombinant pharmaceutical proteins [1,2]
It is important to note that the conditions we reported above are developed for N. benthamiana plants for infiltration with Agrobacterium harboring MagnICON and geminiviral vectors
The results showed that agroinfiltration with geminiviral replicon vectors permits high-level production of VLPs derived from the Norwalk virus capsid protein (NVCP) and therapeutic monoclonal antibodies (mAbs) against Ebola (EBV) or West Nile (WNV) viruses in lettuce [34]
Summary
Plants have been explored as alternatives to mammalian, insect, and bacterial cell cultures for the expression and production of recombinant pharmaceutical proteins [1,2]. Substantial efforts have been focused on the development of efficient transgene delivery methods that are scalable for large-scale transient expression operations, as well as on the search for new expression vectors to further enhance the accumulation of recombinant protein. Expression vector construction, plant growth, Agrobacterium culture preparation, and the actual infiltration are the four major steps of agroinfiltration for the production of pharmaceutical proteins.
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