A novel two-component Tobacco mosaic virus-based vector system for high-level expression of multiple therapeutic proteins including a human monoclonal antibody in plants.
Expression of multiple therapeutic proteins from Tobacco mosaic virus (TMV)-based vectors was not successful when plants were coinoculated with a mixture of two TMV vectors engineered to express two foreign genes individually. Here, we have engineered and developed a defective RNA (dRNA)-based TMV vector (dRT-V) that utilizes two components of the same virus, with the dRNA component depending on the helper virus for replication. Agrobacterium-mediated coinoculation of Nicotiana benthamiana plants with both components of the dRT-V resulted in high-level expression of a human growth hormone and a lichenase-fused lethal factor protein of Bacillus anthracis. Furthermore, both heavy and light chains were expressed and assembled into a monoclonal antibody (mAb) specific to the protective antigen of B. anthracis, and the average yield of the purified antibody obtained was 120 mg/kg of fresh tissue. Our data suggest that dRT-V has a potential for rapid, cost-effective, large-scale manufacturing of multiple therapeutic proteins including mAbs in response to any biological emergencies.
Roy
Gourgopal
G
Fraunhofer USA Center for Molecular Biotechnology, Newark, DE, USA. groy@fraunhofer-cmb.org
Weisburg
Sangeetha
S
Rabindran
Shailaja
S
Yusibov
Vidadi
V
D000907:Antibodies, Bacterial / Q000235:genetics / Q000378:metabolism*; D000911:Antibodies, Monoclonal / Q000378:metabolism*; D005786:Gene Expression Regulation / Q000235:genetics* / Q000502:physiology*; D005818:Genetic Engineering; D005822:Genetic Vectors; D006801:Humans; D014026:Nicotiana / Q000235:genetics* / Q000378:metabolism*; D014027:Tobacco Mosaic Virus
D000907:Antibodies, Bacterial; D000911:Antibodies, Monoclonal