Abstract
Biopharmaceuticals such as antibodies and recombinant subunit vaccines are generally produced on a commercial basis by process-scale fermentation in bacteria, yeast or animal cells. Plants and plant cells have joined the exclusive club of commercial production platforms comparatively recently, but they offer certain advantages over the more established systems particularly in terms of economy, scalability, response times and formulation options. After a promising start and then a rocky transition from early R&D to preclinical and clinical development, plants are now becoming more firmly established as an alternative to microbes and mammalian cells for the production of pharmaceutical proteins. Several plant-derived pharmaceuticals have undergone clinical trials and the first products for human use are approaching market authorization, with antibodies and vaccines strongly represented among these front runners. Although scientific advances have played an important role in the maturation of plant-based production technology, perhaps the most critical development has been the definition of a workable regulatory framework which has recently yielded the first processes for biopharmaceutical production in plants that comply with good manufacturing practice. In this chapter, we consider the state of the art in plant-based production systems for antibodies and vaccines and discus the development issues which remain to be addressed before plants become an acceptable mainstream production technology.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Aviezer D, Brill-Almon E, Shaaltiel Y, Hashmueli S, Bartfeld D, Mizrachi S, Liberman Y, Freeman A, Zimran A, Galun E (2009) A plant-derived recombinant human glucocerebrosidase enzyme – a preclinical and phase I investigation. PLoS One 4:e4792
Cardoso RMF, Zwick MB, Stanfield RL, Kunert R, Binley JM, Katinger H, Burton DR, Wilson IA (2005) Broadly neutralizing anti-HIV antibody, 4E10 recognizes a helical conformation of a highly conserved fusion-associated motif in gp41. Immunity 22:163–173
Castilho A, Strasser R, Stadlmann J, Grass J, Jez J, Gattinger P, Kunert R, Quendler H, Pabst M, Leonard R, Altmann F, Steinkellner H (2010) In planta protein sialylation through overexpression of the respective mammalian pathway. J Biol Chem 285:15923–15930
Conley AJ, Joensuu JJ, Jevnikar AM, Menassa R, Brandle JE (2009) Optimization of elastin-like polypeptide fusions for expression and purification of recombinant proteins in plants. Biotechnol Bioeng 103:562–573
Conley AJ, Joensuu JJ, Richman A, Menassa R (2011) Protein body-inducing fusions for high-level production and purification of recombinant proteins in plants. Plant Biotechnol J 9:419–433
Cox KM, Sterling JD, Regan JT, Gasdaska JR, Frantz KK, Peele CG, Black A, Passmore D, Moldovan-Loomis C, Srinivasan M, Cuison S, Cardarelli PM, Dickey LF (2006) Glycan optimization of a human monoclonal antibody in the aquatic plant Lemna minor. Nat Biotechnol 24:1591–1597
D’Aoust MA, Couture MM, Charland N, Trépanier S, Landry N, Ors F, Vézina LP (2010) The production of hemagglutinin-based virus-like particles in plants: a rapid, efficient and safe response to pandemic influenza. Plant Biotechnol J 8:1–13
De Muynck B, Navarre C, Boutry M (2010) Production of antibodies in plants: status after twenty years. Plant Biotechnol J 8:529–563
Decker EL, Reski R (2004) The moss bioreactor. Curr Opin Plant Biol 7:166–170
Desai P, Shrivastava N, Padh H (2010) Production of heterologous protein in plants: strategies for optimal expression. Biotechnol Adv 28:427–435
Fischer R, Schillberg S, Hellwig S, Twyman RM, Drossard J (2011) GMP issues for plant-derived recombinant proteins. Biotechnol Adv (in press)
Floss DM, Sack M, Stadlmann J, Rademacher T, Scheller J, Stoger E, Fischer R, Conrad U (2008) Biochemical and functional characterization of anti-HIV antibody-ELP fusion proteins from transgenic plants. Plant Biotechnol J 6:379–391
Franklin SE, Mayfield SP (2005) Recent developments in the production of human therapeutic proteins in eukaryotic algae. Expert Opin Biol Ther 5:225–235
Gavilondo JV, Larrick JW (2000) Antibody engineering at the millennium. Biotechniques 29:128–138
Giritch A, Marillonnet S, Engler C, van Eldik G, Botterman J, Klimyuk V, Gleba Y (2006) Rapid high-yield expression of full-size IgG antibodies in plants coinfected with noncompeting viral vectors. Proc Natl Acad Sci USA 103:14701–14706
Gleba Y, Marillonnet S, Klimyuk V (2004) Engineering viral expression vectors for plants: the ‘full virus’ and the ‘deconstructed virus’ strategies. Curr Opin Plant Biol 7:182–188
Gleba Y, Klimyuk V, Marillonnet S (2005) Magnifection – a new platform for expressing recombinant vaccines in plants. Vaccine 23:2042–2048
Gomord V, Sourrouille C, Fitchette AC, Bardor M, Pagny S, Lerouge P, Faye L (2004) Production and glycosylation of plant-made pharmaceuticals: the antibodies as a challenge. Plant Biotechnol J 2:83–100
Gomord V, Fitchette AC, Menu-Bouaouiche L, Saint-Jore-Dupas C, Plasson C, Michaud D, Faye L (2010) Plant-specific glycosylation patterns in the context of therapeutic protein production. Plant Biotechnol J 8:564–587
Gottschalk U (2009) Process scale purification of antibodies. Wiley, New York
Hellwig S, Drossard J, Twyman RM, Fischer R (2004) Plant cell cultures for the production of recombinant proteins. Nat Biotechnol 22:1415–1422
Hiatt A, Cafferkey R, Bowdish K (1989) Production of antibodies in transgenic plants. Nature 342:76–78
Hood EE (2002) From green plants to industrial enzymes. Enzyme Microb Technol 30:279–283
Hood EE, Witcher DR, Maddock S, Meyer T, Baszczynski C, Bailey M, Flynn P, Register J, Marshall L, Bond D, Kulisek E, Kusnadi A, Evangelista R, Nikolov Z, Wooge C, Mehigh RJ, Herman R, Kappel WK, Ritland D, Li CP, Howard J (1997) Commercial production of avidin from transgenic maize: characterization of transformant, production, processing, extraction and purification. Mol Breed 3:291–306
Huang Z, Phoolcharoen W, Lai H, Piensook K, Cardineau G, Zeitlin L, Whaley KJ, Arntzen CJ, Mason HS, Chen Q (2010) High-level rapid production of full-size monoclonal antibodies in plants by a single-vector DNA replicon system. Biotechnol Bioeng 106:9–17
Kapusta J, Modelska A, Figlerowicz M, Pniewski T, Letellier M, Lisowa O, Yusibov V, Koprowski H, Plucienniczak A, Legocki AB (1999) A plant-derived edible vaccine against hepatitis B virus. FASEB J 13:1796–1799
Kusnadi AR, Evangelista RL, Hood EE, Howard JA, Nikolov ZL (1998) Processing of transgenic corn seed and its effect on the recovery of recombinant β-glucuronidase. Biotechnol Bioeng 60:44–52
Landry N, Ward BJ, Trépanier S, Montomoli E, Dargis M, Lapini G, Vézina LP (2010) Preclinical and clinical development of plant-made virus-like particle vaccine against avian H5N1 influenza. PLoS One 5:e15559
Ma JK, Hikmat BY, Wycoff K, Vine ND, Chargelegue D, Yu L, Hein MB, Lehner T (1998) Characterization of a recombinant plant monoclonal secretory antibody and preventive immunotherapy in humans. Nat Med 4:601–606
Ma JKC, Drake P, Christou P (2003) The production of recombinant pharmaceutical proteins in plants. Nat Rev Genet 4:794–805
Ma JKC, Barros E, Bock R, Christou P, Dale PJ, Dix PJ, Fischer R, Irwin J, Mahoney R, Pezzotti M, Schillberg S, Sparrow P, Stoger E, Twyman RM (2005a) Molecular farming for new drugs and vaccines. Current perspectives on the production of pharmaceuticals in transgenic plants. EMBO Rep 6:593–599
Ma JKC, Chikwamba R, Dale PJ, Fischer R, Mahoney R, Twyman RM (2005b) Plant-derived pharmaceuticals – the road forward. Trends Plant Sci 10:580–585
Marillonnet S, Thoeringer C, Kandzia R, Klimyuk V, Gleba Y (2005) Systemic Agrobacterium tumefaciens-mediated transfection of viral replicons for efficient transient expression in plants. Nat Biotechnol 23:718–723
McCormick AA, Kumagai MH, Hanley K, Turpen TH, Hakim I, Grill LK, Tuse D, Levy S, Levy R (1999) Rapid production of specific vaccines for lymphoma by expression of the tumor-derived single-chain Fv epitopes in tobacco plants. Proc Natl Acad Sci USA 96:703–708
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
Menkhaus TJ, Bai Y, Zhang C, Nikolov ZL, Glatz CE (2004) Considerations for the recovery of recombinant proteins from plants. Biotechnol Prog 20:1001–1014
Nikolov ZL, Woodard SL (2004) Downstream processing of recombinant proteins from transgenic feedstock. Curr Opin Biotechnol 15:479–486
Nochi T, Takagi H, Yuki Y, Yang L, Masumura T, Mejima M, Nakanishi U, Matsumura A, Uozumi A, Hiroi T, Morita S, Tanaka K, Takaiwa F, Kiyono H (2007) Rice-based mucosal vaccine as a global strategy for cold-chain- and needle-free vaccination. Proc Natl Acad Sci USA 104:10986–10991
Pogue GP, Vojdani F, Palmer KE, Hiatt E, Hume S, Phelps J, Long L, Bohorova N, Kim D, Pauly M, Velasco J, Whaley K, Zeitlin L, Garger SJ, White E, Bai Y, Haydon H, Bratcher B (2010) Production of pharmaceutical-grade recombinant aprotinin and a monoclonal antibody product using plant-based transient expression systems. Plant Biotechnol J 8:638–654
Rademacher T, Sack M, Arcalis E, Stadlmann J, Balzer S, Altmann F, Quendler H, Stiegler G, Kunert R, Fischer R, Stoger E (2008) Recombinant antibody 2G12 produced in maize endosperm efficiently neutralize HIV-1 and contains predominantly single-GlcNAc N-glycans. Plant Biotechnol J 6:189–201
Ramessar K, Rademacher T, Sack M, Stadlmann J, Platis D, Stiegler G, Labrou N, Altmann F, Ma J, Stöger E, Capell T, Christou P (2008) Cost-effective production of a vaginal protein microbicide to prevent HIV transmission. Proc Natl Acad Sci USA 105:3727–3732
Raskin I, Ribnicky DM, Komarnytsky S, Ilic N, Poulev A, Borisjuk N, Brinker A, Moreno DA, Ripoll C, Yakoby N, O’Neal JM, Cornwell T, Pastor I, Fridlender B (2002) Plants and human health in the twenty-first century. Trends Biotechnol 20:522–531
Richter LJ, Thanavala Y, Arntzen CJ, Mason HS (2000) Production of hepatitis B surface antigen in transgenic plants for oral immunization. Nat Biotechnol 18:1167–1171
Rybicki EP (2010) Plant-made vaccines for humans and animals. Plant Biotechnol J 8:620–637
Sack M, Paetz A, Kunert R, Bomble M, Hesse F, Stiegler G, Fischer R, Katinger H, Stoger E, Rademacher T (2007) Functional analysis of the broadly neutralizing human anti-HIV-1 antibody 2F5 produced in transgenic BY-2 suspension cultures. FASEB J 21:1655–1664
Sainsbury F, Lomonossoff GP (2008) Extremely high-level and rapid protein production in plants without the use of viral replication. Plant Physiol 148:1212–1218
Schähs M, Strasser R, Stadlmann J, Kunert R, Rademacher T, Steinkellner H (2007) Production of a monoclonal antibody in plants with a humanized N-glycosylation pattern. Plant Biotechnol J 5:657–663
Schillberg S, Zimmermann S, Zhang MY, Fischer R (2001) Antibody-based resistance to plant pathogens. Transgenic Res 10:1–12
Schillberg S, Fischer R, Emans N (2003) Molecular farming of recombinant antibodies in plants. Cell Mol Life Sci 60:433–445
Sharma AK, Sharma MK (2009) Plants as bioreactors: recent developments and emerging opportunities. Biotechnol Adv 27:811–832
Sheridan C (2010) Fresh from the biologic pipeline – 2009. Nat Biotechnol 28:307–310
Sijmons PC, Dekker BMM, Schrammeijer B, Verwoerd TC, Van Den Elzen PJM, Hoekema A (1990) Production of correctly processed human serum albumin in transgenic plants. Bio/Technol 8:217–221
Spok A, Twyman RM, Fischer R, Ma JKC, Sparrow PAC (2008) Evolution of a regulatory framework for plant-made pharmaceuticals. Trends Biotechnol 26:506–517
Sriraman R, Bardor M, Sack M, Vaquero C, Faye L, Fischer R, Finnern R, Lerouge P (2004) Recombinant anti-hCG antibodies retained in the endoplasmic reticulum of transformed plants lack core-xylose and core-α(1,3)fucose residues. Plant Biotechnol J 2:279–287
Strasser R, Altmann F, Mach L, Glossl J, Steinkellner H (2004) Generation of Arabidopsis thaliana plants with complex N-glycans lacking β1,2-linked xylose and core α1,3-linked fucose. FEBS Lett 561:132–136
Strasser R, Stadlmann J, Schähs M, Stiegler G, Quendler H, Mach L, Glössl J, Weterings K, Pabst M, Steinkellner H (2008) Generation of glycoengineered Nicotiana benthamiana for the production of monoclonal antibodies with a homogeneous human-like N-glycan structure. Plant Biotechnol J 6:392–402
Strasser R, Castilho A, Stadlmann J, Kunert R, Quendler H, Gattinger P, Jez J, Rademacher T, Altmann F, Mach L, Steinkellner H (2009) Improved virus neutralization by plant-produced anti-HIV antibodies with a homogeneous β1,4-galactosylated N-glycan profile. J Biol Chem 284:20479–20485
Tacket CO, Mason HS, Losonsky G, Clements JD, Levine MM, Arntzen CJ (1998) Immunogenicity in humans of a recombinant bacterial-antigen delivered in transgenic potato. Nat Med 4:607–609
Tacket CO, Mason HS, Losonsky G, Estes MK, Levine MM, Arntzen CJ (2000) Human immune responses to a novel Norwalk virus vaccine delivered in transgenic potatoes. J Infect Dis 182:302–305
Tacket CO, Pasetti MF, Edelman R, Howard JA, Streatfield S (2004) Immunogenicity of recombinant LT-B delivered orally to humans in transgenic corn. Vaccine 22:4385–4389
Tiwari S, Verma PC, Singh PK, Tuli R (2009) Plants as bioreactors for the production of vaccine antigens. Biotechnol Adv 27:449–467
Torrent M, Llompart B, Lasserre-Ramassamy S, Llop-Tous I, Bastida M, Marzabal P, Westerholm-Parvinen A, Saloheimo M, Heifetz PB, Ludevid MD (2009) Eukaryotic protein production in designed storage organelles. BMC Biol 7:5
Triguero A, Cabrera G, Cremata JA, Yuen CT, Wheeler J, RamÃrez NI (2005) Plant-derived mouse IgG monoclonal antibody fused to KDEL endoplasmic reticulum-retention signal is N-glycosylated homogeneously throughout the plant with mostly high-mannose-type N-glycans. Plant Biotechnol J 3:449–457
Twyman RM, Stoger E, Schillberg S, Christou P, Fischer R (2003) Molecular farming in plants: host systems and expression technology. Trends Biotechnol 21:570–578
Twyman RM, Schillberg S, Fischer R (2005) Transgenic plants in the biopharmaceutical market. Expert Opin Emerg Drugs 10:185–218
Vézina LP, Faye L, Lerouge P, D’Aoust MA, Marquet-Blouin E, Burel C, Lavoie PO, Bardor M, Gomord V (2009) Transient co-expression for fast and high-yield production of antibodies with human-like N-glycans in plants. Plant Biotechnol J 7:442–455
Whaley KJ, Hiatt A, Zeitlin L (2011) Emerging antibody products and Nicotiana manufacturing. Hum Vaccine 7:349–356
Witcher D, Hood EE, Peterson D, Bailey M, Bond D, Kusnadi A, Evangelista R, Nikolov Z, Wooge C, Mehigh R, Kappel W, Register JC, Howard JA (1998) Commercial production of β-glucuronidase (GUS): a model system for the production of proteins in plants. Mol Breed 4:301–312
Xu J, Ge X, Dolan MC (2011) Towards high-yield production of pharmaceutical proteins with plant cell suspension cultures. Biotechnol Adv 29:278–299
Yusibov V, Hooper DC, Spitsin SV, Fleysh N, Kean RB, Mikheeva T, Deka D, Karasev A, Cox S, Randall J, Koprowski H (2002) Expression in plants and immunogenicity of plant virus-based experimental rabies vaccine. Vaccine 20:3155–3164
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2012 Springer Science+Business Media B.V.
About this chapter
Cite this chapter
Twyman, R.M., Schillberg, S., Fischer, R. (2012). The Production of Vaccines and Therapeutic Antibodies in Plants. In: Wang, A., Ma, S. (eds) Molecular Farming in Plants: Recent Advances and Future Prospects. Springer, Dordrecht. https://doi.org/10.1007/978-94-007-2217-0_7
Download citation
DOI: https://doi.org/10.1007/978-94-007-2217-0_7
Published:
Publisher Name: Springer, Dordrecht
Print ISBN: 978-94-007-2216-3
Online ISBN: 978-94-007-2217-0
eBook Packages: Chemistry and Materials ScienceChemistry and Material Science (R0)