Rapid Transient Production in Plants by Replicating and Non-Replicating Vectors Yields High Quality Functional Anti-HIV Antibody

Sainsbury, Frank, Sack, Markus, Stadlmann, Johannes, Quendler, Heribert, Fischer, Rainer and Lomonossoff, George P. (2010) Rapid Transient Production in Plants by Replicating and Non-Replicating Vectors Yields High Quality Functional Anti-HIV Antibody. PLoS One, 5 11: . doi:10.1371/journal.pone.0013976


Author Sainsbury, Frank
Sack, Markus
Stadlmann, Johannes
Quendler, Heribert
Fischer, Rainer
Lomonossoff, George P.
Title Rapid Transient Production in Plants by Replicating and Non-Replicating Vectors Yields High Quality Functional Anti-HIV Antibody
Journal name PLoS One   Check publisher's open access policy
ISSN 1932-6203
Publication date 2010-11-01
Year available 2010
Sub-type Article (original research)
DOI 10.1371/journal.pone.0013976
Open Access Status DOI
Volume 5
Issue 11
Total pages 9
Place of publication San Francisco, CA United States
Publisher Public Library of Science
Language eng
Formatted abstract
Background: 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.

Methodology/Principal Findings: 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.

Conclusions: 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.
 
Keyword Cowpea mosaic virus
Dna Replicon System
Monoclonal antibody
Viral Vectors
Neutralizing Antibodies
Endoplasmic Reticulum
Q-Index Code C1
Q-Index Status Provisional Code
Grant ID MEST-CT-2004-504273
Institutional Status Non-UQ

Document type: Journal Article
Sub-type: Article (original research)
Collection: Australian Institute for Bioengineering and Nanotechnology Publications
 
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