2006 AIChE Annual Meeting
(425h) Transient Expression of Functional Human Alpha-1-Antitrypsin in Nicotiana benthamiana Plants and Suspension Cultures
Authors
Michael A. Plesha - Presenter, University of California, Davis
Ting-Kuo Huang, University of California, Davis
Mysore R. Sudarshana, University of California, Davis
Sandra L. Uratsu, University of California, Davis
Bryce W. Falk, University of California at Davis
Abhaya M. Dandekar, University of California
Alan P. Jackman, University of California, Davis
Karen A. McDonald, University of California at Davis
Transient recombinant protein expression has been evaluated as a production route capable of enabling rapid, large-scale production of functional human therapeutic proteins that could be needed for widespread treatment and/or vaccination in the event of a crisis. In this study, transient expression of a model human blood protein, alpha-1-antitrypsin (AAT), was demonstrated by agroinfiltration of Nicotiana benthamiana leaves and suspension cultures. Production of functional recombinant human AAT was evaluated in planta after coinfiltration with Agrobacterium tumefaciens carrying one of three different expression cassettes (constitutive, chemically inducible, or a chemically-inducible viral amplicon) and Agrobacterium tumefaciens carrying the potent gene silencing suppressor p19. Functional recombinant AAT reached levels of up to 17 mg per 100 g leaf tissue and was stable in plant biomass for up to four days. Production in planta was compared to recombinant AAT produced constitutively in suspension cultures. By utilizing this transient expression system, functional human protein was produced in just 4 days from a frozen stock of recombinant Agrobacterium tumefaciens.