Abstract
Variability in recombinant IgG yield in transgenic tobacco plants has previously been observed in relation to leaf position, and is interpreted as a function of ageing and the senescence process, leading to increasing protein degradation. Here, similar findings are demonstrated in plants of different ages, expressing IgG but not IgG-HDEL, an antibody form that accumulates within the endoplasmic reticulum. Antibody yields declined following wounding in young transgenic plants expressing IgG but not in those expressing IgG-HDEL. However, in mature IgG plants, the opposite was demonstrated, with significant boosts in yield, while mature IgG-HDEL plants could not be boosted. The lack of response in IgG-HDEL plants suggests that the changes induced by wounding occur post-translationally, and the findings might be explained by wounding responses that differ in plants according to their developmental stages. Plant mechanisms involved in senescence and wounding overlap to a significant degree and compounds such as ethylene, jasmonic acid and salicylic acid are important for mediating downstream effects. Treatment of transgenic plants with ethylene also resulted in a decrease in recombinant IgG yield, which was consistent with the finding that wounded plants could induce lower IgG yields in neighbouring non-wounded plants. Treatment with 1-MCP, an ethylene antagonist, abrogated the IgG yield drop that resulted from wounding, but had no effect on the more gradual IgG yield loss associated with increasing plant age.
Similar content being viewed by others
References
Bowles D (1998) Signal transduction in the wound response of tomato plants. Philos Trans R Soc Lond B Biol Sci 353:1495–1510
Buchanan-Wollaston V, Page T, Harrison E, Breeze E, Lim PO, Nam HG, Lin JF, Wu SH, Swidzinski J, Ishizaki K, Leaver CJ (2005) Comparative transcriptome analysis reveals significant differences in gene expression and signalling pathways between developmental and dark/starvation-induced senescence in Arabidopsis. Plant J 42:567–585
Chang C, Kwok SF, Bleecker AB, Meyerowitz EM (1993) Arabidopsis ethylene-response gene Etr1—similarity of product to 2-component regulators. Science 262:539–544
Colgan R, Atkinson CJ, Paul M, Hassan S, Drake PMW, Sexton AL, Santa-Cruz S, James D, Hamp K, Gutteridge C and Ma JK-C (2010) Optimisation of contained Nicotiana tabacum cultivation for the production of recombinant protein pharmaceuticals. Transgenic Res 19(2):241–256
Colleluori DM, Tien D, Kang F, Pagliei T, Kuss R, McCormick T, Watson K, McFadden K, Chaiken I, Buckheit RW Jr, Romano JW (2005) Expression, purification, and characterization of recombinant cyanovirin-N for vaginal anti-HIV microbicide development. Protein Expr Purif 39:229–236
Conrad U, Fiedler U (1998) Compartment-specific accumulation of recombinant immunoglobulins in plant cells: an essential tool for antibody production and immunomodulation of physiological functions and pathogen activity. Plant Mol Biol 38:101–109
Elbers IJ, Stoopen GM, Bakker H, Stevens LH, Bardor M, Molthoff JW, Jordi WJ, Bosch D, Lommen A (2001) Influence of growth conditions and developmental stage on N-glycan heterogeneity of transgenic immunoglobulin G and endogenous proteins in tobacco leaves. Plant Physiol 126:1314–1322
Farmer EE, Ryan CA (1990) Interplant communication—airborne methyl jasmonate induces synthesis of proteinase-inhibitors in plant-leaves. Proc Natl Acad Sci USA 87:7713–7716
Gepstein S (2004) Leaf senescence—not just a ‘wear and tear’ phenomenon. Genome Biol 5:2
Gepstein S, Sabehi G, Carp MJ, Hajouj T, Nesher MFO, Yariv I, Dor C, Bassani M (2003) Large-scale identification of leaf senescence-associated genes. Plant J 36:629–642
Gilissen LJW, vanStaveren MJ, Hakkert JC, Smulders MJM (1996) Competence for regeneration during tobacco internodal development - Involvement of plant age, cell elongation stage, and degree of polysomaty. Plant Physiol 111:1243–1250
Green TR, Ryan CA (1972) Wound-Induced Proteinase Inhibitor in Plant Leaves - Possible Defense Mechanism against Insects. Science 175:776–777
Hassan S, van Dolleweerd CJ, Ioakeimidis F, Keshavarz-Moore E, Ma JK (2008) Considerations for extraction of monoclonal antibodies targeted to different subcellular compartments in transgenic tobacco plants. Plant Biotechnol J 6:733–748
Herms DA, Mattson WJ (1992) The dilemma of plants—to grow or defend. Q Rev Biol 67:283–335
Ma JK, Lehner T, Stabila P, Fux CI, Hiatt A (1994) Assembly of monoclonal antibodies with IgG1 and IgA heavy chain domains in transgenic tobacco plants. Eur J Immunol 24:131–138
McCormick AA, Reddy S, Reinl SJ, Cameron TI, Czerwinkski DK, Vojdani F, Hanley KM, Garger SJ, White EL, Novak J, Barrett J, Holtz RB, Tuse D, Levy R (2008) Plant-produced idiotype vaccines for the treatment of non-Hodgkin’s lymphoma: safety and immunogenicity in a phase I clinical study. Proc Natl Acad Sci USA 105:10131–10136
Muller T, Dietzschold B, Ertl H, Fooks AR, Freuling C, Fehlner-Gardiner C, Kliemt J, Meslin FX, Franka R, Rupprecht CE, Tordo N, Wanderler AI, Kieny MP (2009) Development of a mouse monoclonal antibody cocktail for post-exposure rabies prophylaxis in humans. PLoS Negl Trop Dis 3:e542
Nuttall J, Vine N, Hadlington JL, Drake P, Frigerio L, Ma JK (2002) ER-resident chaperone interactions with recombinant antibodies in transgenic plants. Eur J Biochem 269:6042–6051
ODonnell PJ, Calvert C, Atzorn R, Wasternack C, Leyser HMO, Bowles DJ (1996) Ethylene as a signal mediating the wound response of tomato plants. Science 274:1914–1917
Schilmiller AL, Howe GA (2005) Systemic signaling in the wound response. Curr Opin Plant Biol 8:369–377
Sexton A, Drake PM, Mahmood N, Harman SJ, Shattock RJ, Ma JK (2006) Transgenic plant production of Cyanovirin-N, an HIV microbicide. FASEB J 20:356–358
Shi QM, Li CJ, Zhang FS (2006) Nicotine synthesis in Nicotiana tabacum L. induced by mechanical wounding is regulated by auxin. J Exp Bot 57:2899–2907
Sisler EC, Serek M, Dupille E (1996) Comparison of cyclopropene, 1-methylcyclopropene, and 3,3-dimethylcyclopropene as ethylene antagonists in plants. Plant Growth Regul 18:169–174
Sparrow PAC, Irwin JA, Dale PJ, Twyman RM, Ma JKC (2007) Pharma-planta: road testing the developing regulatory guidelines for plant-made pharmaceuticals. Transgenic Res 16:147–161
Stevens LH, Stoopen GM, Elbers IJ, Molthoff JW, Bakker HA, Lommen A, Bosch D, Jordi W (2000) Effect of climate conditions and plant developmental stage on the stability of antibodies expressed in transgenic tobacco. Plant Physiol 124:173–182
Valdes R, Gomez L, Padilla S, Brito J, Reyes B, Alvarez T, Mendoza O, Herrera O, Ferro W, Pujol M, Leal V, Linares M, Hevia Y, Garcia C, Mila L, Garcia O, Sanchez R, Acosta A, Geada D, Paez R, Luis VJ, Borroto C (2003) Large-scale purification of an antibody directed against hepatitis B surface antigen from transgenic tobacco plants. Biochem Biophys Res Commun 308:94–100
van Dolleweerd CJ, Chargelegue D, Ma JK (2003) Characterization of the conformational epitope of Guy’s 13, a monoclonal antibody that prevents Streptococcus mutans colonization in humans. Infect Immun 71:754–765
Wu J, Hettenhausen C, Meldau S, Baldwin IT (2007) Herbivory rapidly activates MAPK signaling in attacked and unattacked leaf regions but not between leaves of Nicotiana attenuata. Plant Cell 19:1096–1122
Acknowledgments
The authors would like to acknowledge DTI (TP/3/BIO/6/I/17346), the EU Pharma-Planta Integrated Project and the Hotung Foundation for supporting the project. Also, Dr. Barry O’Keefe for providing CV-N specific antiserum, and Dr. Gillian Arnold for statistical support. We also thank Mike Davies for technical support at EMR and Dr. Vicky Buchanan-Wollaston for helpful discussions.
Author information
Authors and Affiliations
Corresponding author
Additional information
Sally Hassan and Richard Colgan contributed equally to the work.
Electronic supplementary material
Below is the link to the electronic supplementary material.
Rights and permissions
About this article
Cite this article
Hassan, S., Colgan, R., Paul, M.J. et al. Recombinant monoclonal antibody yield in transgenic tobacco plants is affected by the wounding response via an ethylene dependent mechanism. Transgenic Res 21, 1221–1232 (2012). https://doi.org/10.1007/s11248-012-9595-1
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s11248-012-9595-1