High‐level rapid production of full‐size monoclonal antibodies in plants by a single‐vector DNA replicon system
- 31 December 2009
- journal article
- research article
- Published by Wiley in Biotechnology & Bioengineering
- Vol. 106 (1), 9-17
- https://doi.org/10.1002/bit.22652
Abstract
Plant viral vectors have great potential in rapid production of important pharmaceutical proteins. However, high‐yield production of hetero‐oligomeric proteins that require the expression and assembly of two or more protein subunits often suffers problems due to the “competing” nature of viral vectors derived from the same virus. Previously we reported that a bean yellow dwarf virus (BeYDV)‐derived, three‐component DNA replicon system allows rapid production of single recombinant proteins in plants (Huang et al., 2009. Biotechnol Bioeng 103: 706–714). In this article, we report further development of this expression system for its application in high‐yield production of oligomeric protein complexes including monoclonal antibodies (mAbs) in plants. We showed that the BeYDV replicon system permits simultaneous efficient replication of two DNA replicons and thus, high‐level accumulation of two recombinant proteins in the same plant cell. We also demonstrated that a single vector that contains multiple replicon cassettes was as efficient as the three‐component system in driving the expression of two distinct proteins. Using either the non‐competing, three‐vector system or the multi‐replicon single vector, we produced both the heavy and light chain subunits of a protective IgG mAb 6D8 against Ebola virus GP1 (Wilson et al., 2000. Science 287: 1664–1666) at 0.5 mg of mAb per gram leaf fresh weight within 4 days post‐infiltration of Nicotiana benthamiana leaves. We further demonstrated that full‐size tetrameric IgG complex containing two heavy and two light chains was efficiently assembled and readily purified, and retained its functionality in specific binding to inactivated Ebola virus. Thus, our single‐vector replicon system provides high‐yield production capacity for hetero‐oligomeric proteins, yet eliminates the difficult task of identifying non‐competing virus and the need for co‐infection of multiple expression modules. The multi‐replicon vector represents a significant advance in transient expression technology for antibody production in plants. Biotechnol. Bioeng. 2010; 106: 9–17.Keywords
This publication has 29 references indexed in Scilit:
- A DNA replicon system for rapid high‐level production of virus‐like particles in plantsBiotechnology & Bioengineering, 2009
- Extremely High-Level and Rapid Transient Protein Production in Plants without the Use of Viral ReplicationPlant Physiology, 2008
- An efficient plant viral expression system generating orally immunogenic Norwalk virus-like particlesVaccine, 2008
- Production of vaccines and therapeutic antibodies for veterinary applications in transgenic plants: an overviewTransgenic Research, 2007
- Rapid high-yield expression of full-size IgG antibodies in plants coinfected with noncompeting viral vectorsProceedings of the National Academy of Sciences, 2006
- Use of viral vectors for vaccine production in plantsImmunology & Cell Biology, 2005
- Fluorescent labelling reveals spatial separation of potyvirus populations in mixed infected Nicotiana benthamiana plantsJournal of General Virology, 2003
- The production of recombinant pharmaceutical proteins in plantsNature Reviews Genetics, 2003
- Monoclonal antibody manufacturing in transgenic plants — myths and realitiesCurrent Opinion in Biotechnology, 2002
- Production of antibodies in transgenic plantsNature, 1989