Gene Delivery into Plant Cells for Recombinant Protein Production

Abstract

Recombinant proteins are primarily produced from cultures of mammalian, insect, and bacteria cells. In recent years, the development of deconstructed virus-based vectors has allowed plants to become a viable platform for recombinant protein production, with advantages in versatility, speed, cost, scalability, and safety over the current production paradigms. In this paper, we review the recent progress in the methodology of agroinfiltration, a solution to overcome the challenge of transgene delivery into plant cells for large-scale manufacturing of recombinant proteins. General gene delivery methodologies in plants are first summarized, followed by extensive discussion on the application and scalability of each agroinfiltration method. New development of a spray-based agroinfiltration and its application on field-grown plants is highlighted. The discussion of agroinfiltration vectors focuses on their applications for producing complex and heteromultimeric proteins and is updated with the development of bridge vectors. Progress on agroinfiltration in Nicotiana and non-Nicotiana plant hosts is subsequently showcased in context of their applications for producing high-value human biologics and low-cost and high-volume industrial enzymes. These new advancements in agroinfiltration greatly enhance the robustness and scalability of transgene delivery in plants, facilitating the adoption of plant transient expression systems for manufacturing recombinant proteins with a broad range of applications.

Gene-silencing suppressors for high-level production of the HIV-1 entry inhibitor griffithsin in Nicotiana benthamiana

“…In addition, transient expression is genome integration-independent strategy, which will not be affected by position effects existing in stable transformation, once that the expression vector remains an episomal DNA molecule [22]. Our result is in accordance with previous works, which indicated that agroinfiltration could efficiently resulted in transferring of Agrobacterium into the plant leaves and robust expression of recombinant proteins [24,[52][53][54]. In our study, the infiltration of agrobacterium into the intercellular space of the leaf was performed with syringe method (Fig.…”

“…Moreover, lower induction of the immune system and no limitation in terms of the size of transgenic DNA add versatility to this system. In addition to its speed and high yield, the transient expression platform presents versatility in terms of the expression and accumulation of personalized recombinant proteins, including therapeutics for patient-specific cancers and vaccines for viruses that show rapid antigenic drift and/or multiple strains with unpredictable outbreaks [24,25].…”

Using transgenic plants and modified plant viruses for the development of treatments for human diseases

“…The peak protein expression is typically observed in less than 7 days postinfiltration which is significantly faster when compared to the full virus strategy which requires more than 2 weeks in order to generate a systemic infection for expression. The promise of this platform has been evidenced in numerous successful clinical trials, which demonstrated safety and efficacy of plant-made protein therapeutics and biologics [11 ]. For example, in responding to the H1N1 influenza virus pandemic that occurred in 2009, Medicago, a Canadian company, reported producing the vaccine candidate, hemagglutinin in 19 days in Nicotiana benthamiana [10].…”

Transient Expression of an Immunogenic Envelope Attachment Glycoprotein of Nipah Virus in Nicotiana benthamiana

“…Plant transient expression can be used to produce vaccines at low cost on an agricultural scale and with a low risk of contamination of the product with animal pathogens as reviewed in (Chen & Lai 2015). Furthermore, plant-based vaccines have been proposed as a new approach for oral delivery such as expression of recombinant protein in edible plants like corn and rice (Kapila et al 1997;Nochi et al 2007;Shoji et al 2011).…”