Agroinfiltration as an Effective and Scalable Strategy of Gene Delivery for Production of Pharmaceutical Proteins

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.

Generation and Analysis of Novel Plant-Derived Antibody-Based Therapeutic Molecules against West Nile Virus

“…The resulting coding sequence of pHu-E16scFv-C H 1-3 was cloned into a MagnICON-based plant expression vector [13], and transformed into Agrobacterium tumefaciens . A. tumefaciens strains containing the pHu-E16scFv-C H 1-3 construct were co-delivered into N. benthamiana leaves along with the promoter module and an integrase construct through agroinfiltration [14], [15]. Western blot analysis after reducing or non-reducing gel electrophoresis confirmed that pHu-E16scFv-C H 1-3 was produced in leaves with the expected molecular weight ( Fig 1B , Lane 5 ), and that it assembled into a dimer ( Fig 1D , Lane 7 ).…”

In vitro plant tissue culture: means for production of biological active compounds

“…Bioballistics can be used on a wider range of plant genotypes than are amenable to Agrobacterium transformation. This technique lacks the pathogenic characteristic of the bacteria and simplifies the cloning process since it does not require a specific vector (Chen et al 2014b;Altpeter et al 2016). However, plant tissues subjected to this technique often show difficulties in regeneration after bombardment and in the transgene performance.…”

Optimization of inside and outside factors to improve recombinant protein yield in plant

“…A recent significant development in the area of agroinfiltration is using "deconstructed" viral vectors. In this new type of vector, unnecessary viral genome components for the function of plasmid expression are removed, which leads to the assembly of larger transgenes while keeping viral replication and transcription (Chen et al 2013a). The MagnICON system represents an efficient and robust genetransferring technology for the transient expression of biopharmaceuticals in plant platforms.…”