Rapid Transient Production in Plants by Replicating and Non-Replicating Vectors Yields High Quality Functional Anti-HIV Antibody
Open Access
- 12 November 2010
- journal article
- research article
- Published by Public Library of Science (PLoS) in PLOS ONE
- Vol. 5 (11), e13976
- https://doi.org/10.1371/journal.pone.0013976
Abstract
The capacity of plants and plant cells to produce large amounts of recombinant protein has been well established. Due to advantages in terms of speed and yield, attention has recently turned towards the use of transient expression systems, including viral vectors, to produce proteins of pharmaceutical interest in plants. However, the effects of such high level expression from viral vectors and concomitant effects on host cells may affect the quality of the recombinant product. To assess the quality of antibodies transiently expressed to high levels in plants, we have expressed and characterised the human anti-HIV monoclonal antibody, 2G12, using both replicating and non-replicating systems based on deleted versions of Cowpea mosaic virus (CPMV) RNA-2. The highest yield (approximately 100 mg/kg wet weight leaf tissue) of affinity purified 2G12 was obtained when the non-replicating CPMV-HT system was used and the antibody was retained in the endoplasmic reticulum (ER). Glycan analysis by mass-spectrometry showed that the glycosylation pattern was determined exclusively by whether the antibody was retained in the ER and did not depend on whether a replicating or non-replicating system was used. Characterisation of the binding and neutralisation properties of all the purified 2G12 variants from plants showed that these were generally similar to those of the Chinese hamster ovary (CHO) cell-produced 2G12. Overall, the results demonstrate that replicating and non-replicating CPMV-based vectors are able to direct the production of a recombinant IgG similar in activity to the CHO-produced control. Thus, a complex recombinant protein was produced with no apparent effect on its biochemical properties using either high-level expression or viral replication. The speed with which a recombinant pharmaceutical with excellent biochemical characteristics can be produced transiently in plants makes CPMV-based expression vectors an attractive option for biopharmaceutical development and production.This publication has 59 references indexed in Scilit:
- High‐level rapid production of full‐size monoclonal antibodies in plants by a single‐vector DNA replicon systemBiotechnology & Bioengineering, 2009
- pEAQ: versatile expression vectors for easy and quick transient expression of heterologous proteins in plantsPlant Biotechnology Journal, 2009
- Transient co‐expression for fast and high‐yield production of antibodies with human‐like N‐glycans in plantsPlant Biotechnology Journal, 2009
- 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
- Cost-effective production of a vaginal protein microbicide to prevent HIV transmissionProceedings of the National Academy of Sciences of the United States of America, 2008
- TRBO: A High-Efficiency Tobacco Mosaic Virus RNA-Based Overexpression VectorPlant Physiology, 2007
- Rapid high-yield expression of full-size IgG antibodies in plants coinfected with noncompeting viral vectorsProceedings of the National Academy of Sciences of the United States of America, 2006
- Use of viral vectors for vaccine production in plantsImmunology & Cell Biology, 2005
- Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4Nature, 1970