ISPE San Diego Chapter, Lucas Arzola, Graduate Student, University of California San Diego
The 2001 anthrax mailings raised awareness of the devastating effect a massive bioterrorist attack could have. In case of a large-scale outbreak, responsive, scalable, and cost-effective manufacturing technologies are needed to rapidly deliver effective therapeutics into the clinical setting. Non-transgenic tobacco plants can be used as cost-effective biofactories of therapeutic proteins. Agrobacterium, a bacteria that naturally transfers genetic material into plants, can be engineered to rapidly enable production of a protein of interest in tobacco. The infiltration of Agrobacterium into tobacco comprises the innovative manufacturing platform we are developing. Our research focuses on developing this technology for producing CMG2-Fc, an experimental antibody for anthrax. Of particular interest is using suppression of plant gene silencing as a strategy to increase the yield of CMG2-Fc in tobacco. We tested the co-production of CMG2-Fc with nine different gene silencing suppressors: p1, p10, p19, p21, p24, p25, p38, 2b, and HCPro. The ELISA assay was performed to quantitatively measure the amount of CMG2-Fc protein produced in tobacco. We observed that using the gene silencing suppressor p1 resulted in a maximum production of 0.56 g CMG2-Fc per kg of tobacco leaf after only 3.5 days post-infiltration, a ten-fold increase in the production level when compared to absence of suppression. This result is significant because 1 gram of protein per kilogram of leaf is considered the production threshold that will make this technology commercially viable. This innovative manufacturing platform has great promise, because it is easy to customize for quick and large-scale production of virtually any therapeutic protein, including vaccines and antibodies.
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