Skip to main content

Thank you for visiting You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

Genetic modification

The production of recombinant pharmaceutical proteins in plants

Key Points

  • The use of plants as expression hosts for the large-scale production of recombinant proteins is a recent innovation that has potential advantages of economy, scalability and safety over traditional expression systems.

  • Many pharmaceutical proteins have been expressed in plants as part of 'proof of principle' studies, including human and animal proteins, recombinant subunit vaccines and recombinant antibodies. Only a few of these proteins have reached advanced stages of development and even fewer have begun clinical trials.

  • Most pharmaceutical proteins have been produced in transgenic tobacco plants, because tobacco has a long history as a model organism and robust expression constructs are available. However, there is increasing interest in the use of other species, particularly cereals, legumes, fruit and vegetables.

  • Diverse plant-based expression systems, such as transient expression, plant cell-suspension cultures, recombinant plant viruses and the chloroplast transgenic system are being investigated.

  • Pharmaceutical proteins that are expressed in dry cereal and legume seeds are highly stable and can be stored for long periods at room temperature with no loss of activity.

  • The expression of antibodies and vaccines in edible fruit and vegetables might allow oral administration in partially processed plant tissues.

  • There are some differences between the glycan structures of recombinant glycoproteins that are produced in animals and plants, but so far there is no evidence that such differences cause adverse reactions in human patients.

  • The acceptability of plant-derived pharmaceutical proteins depends on the production of such proteins under cGMP conditions, in line with other expression systems.


Imagine a world in which any protein, either naturally occurring or designed by man, could be produced safely, inexpensively and in almost unlimited quantities using only simple nutrients, water and sunlight. This could one day become reality as we learn to harness the power of plants for the production of recombinant proteins on an agricultural scale. Molecular farming in plants has already proven to be a successful way of producing a range of technical proteins. The first plant-derived recombinant pharmaceutical proteins are now approaching commercial approval, and many more are expected to follow.

Access options

Rent or Buy article

Get time limited or full article access on ReadCube.


All prices are NET prices.

Figure 1: Complex long-chain glycan structure in plants and humans.


  1. 1

    Schwartz, J. R. Advances in Escherichia coli production of therapeutic proteins. Curr. Opin. Biotechnol. 12, 195–201 (2001).

    Google Scholar 

  2. 2

    Chu, L. & Robinson, D. K. Industrial choices for protein production by large-scale cell culture. Curr. Opin. Biotechnol. 12, 180–187 (2001).

    CAS  PubMed  Google Scholar 

  3. 3

    Houdebaine, L. M. Transgenic animal bioreactors. Transgenic Res. 9, 305–320 (2000).

    Google Scholar 

  4. 4

    Fischer, R. & Emans, N. Molecular farming of pharmaceutical proteins. Transgenic Res. 9, 279–299 (2000).

    CAS  PubMed  Google Scholar 

  5. 5

    Giddings, G. Transgenic plants as protein factories. Curr. Opin. Biotechnol. 12, 450–454 (2001).

    CAS  PubMed  Google Scholar 

  6. 6

    Barta, A. et al. The expression of a nopaline synthase human growth hormone chimaeric gene in transformed tobacco and sunflower callus tissue. Plant Mol. Biol. 6, 347–357 (1986).

    CAS  PubMed  Google Scholar 

  7. 7

    Hiatt, A., Cafferkey, R. & Bowdish, K. Production of antibodies in transgenic plants. Nature 342, 76–78 (1989).

    CAS  PubMed  Google Scholar 

  8. 8

    Mason, H. S., Lam, D. M. K. & Arntzen, C. J. Expression of hepatitis B surface antigen in transgenic plants. Proc. Natl Acad. Sci. USA 89, 11745–11749 (1992).

    CAS  PubMed  Google Scholar 

  9. 9

    Thanavala, Y., Yang, Y. -F., Lyons, P., Mason, H. S. & Arntzen, C. J. Immunogenicity of transgenic plant-derived hepatitis B surface antigen. Proc. Natl Acad. Sci. USA 92, 3358–3361 (1995).

    CAS  PubMed  Google Scholar 

  10. 10

    Hood, E. E. et al. Criteria for high-level expression of a fungal laccase gene in transgenic maize. Plant Biotechnol. J. 1, 129–140 (2003).

    CAS  PubMed  Google Scholar 

  11. 11

    Hood, E. E. et al. Commercial production of avidin from transgenic maize: characterization of transformant, production, processing, extraction and purification. Mol. Breeding 3, 291–306 (1997).

    CAS  Google Scholar 

  12. 12

    Chong, D. K. X. et al. Expression of the human milk protein β-casein in transgenic potato plants. Transgenic Res. 6, 289–296 (1997).

    CAS  PubMed  Google Scholar 

  13. 13

    Ruggiero, F. et al. Triple helix assembly and processing of human collagen produced in transgenic tobacco plants. FEBS Lett. 469, 132–136 (2000).

    CAS  PubMed  Google Scholar 

  14. 14

    Staub, J. M. et al. High-yield production of a human therapeutic protein in tobacco chloroplasts. Nature Biotechnol. 18, 333–338 (2000). This report shows that the secreted human protein somatotropin is soluble and biologically active when expressed in tobacco chloroplasts and has correctly formed disulphide bonds.

    CAS  Google Scholar 

  15. 15

    Fernandez-San Millan, A., Mingo-Castel, A., Miller, M. & Daniell, H. A chloroplast transgenic approach to hyper-express and purify human serum albumin, a protein highly susceptible to proteolytic degradation. Plant Biotechnol. 1, 77–79 (2003).

    Google Scholar 

  16. 16

    Moloney, M., Boothe, J. & Van Rooijen, G. Oil bodies and associated proteins as affinity matrices. US Patent 6,509,453 (2003)

  17. 17

    Chadd, H. E. & Chamow, S. M. Therapeutic antibody expression technology. Curr. Opin. Biotechnol. 12, 188–194 (2001).

    CAS  PubMed  Google Scholar 

  18. 18

    Ma, J. K. et al. Generation and assembly of secretory antibodies in plants. Science 268, 716–719 (1995). This study shows for the first time that secretory antibodies, with 10 polypeptide chains that represent the products of four genes, can be assembled correctly in transgenic plants. Two rounds of crossing, which involved four singly-transgenic lines, were required to stack all four transgenes in the same plant.

    CAS  PubMed  Google Scholar 

  19. 19

    Richter, L. J., Thanavala, Y., Arntzen, C. J. & Mason, H. S. Production of hepatitis B surface antigen in transgenic plants for oral immunization. Nature Biotechnol. 18, 1167–1171 (2000).

    CAS  Google Scholar 

  20. 20

    Kapusta, J. et al. A plant-derived edible vaccine against hepatitis B virus. FASEB J. 13, 1796–1799 (1999).

    CAS  PubMed  Google Scholar 

  21. 21

    Tacket, C. O. et al. Immunogenicity in humans of a recombinant bacterial-antigen delivered in a transgenic potato. Nature Med. 4, 607–609 (1998).

    CAS  PubMed  Google Scholar 

  22. 22

    Tacket, C. O. et al. Human immune responses to a novel Norwalk virus vaccine delivered in transgenic potatoes. J. Infect. Dis. 182, 302–305 (2000).

    CAS  PubMed  Google Scholar 

  23. 23

    Chong, D. K. X. & Langridge, W. H. R. Expression of full-length bioactive antimicrobial human lactoferrin in potato plants. Transgenic Res. 9, 71–78 (2000).

    CAS  PubMed  Google Scholar 

  24. 24

    Zhang, X., Urry, D. W. & Daniell, H. Expression of an environmentally friendly synthetic protein-based polymer in transgenic tobacco plants. Plant Cell Reps. 16, 174–179 (1996).

    Google Scholar 

  25. 25

    Guda, C., Lee, S. B. & Daniell, H. Stable expression of a biodegradable protein-based polymer in stable tobacco chloroplasts. Plant Cell Reps. 19, 257–262 (2000).

    CAS  Google Scholar 

  26. 26

    Merle, C. et al. Hydroxylated human homotrimeric collagen I in Agrobacterium tumefaciens-mediated transient expression and in transgenic tobacco plant. FEBS Lett. 515, 114–118 (2002).

    CAS  PubMed  Google Scholar 

  27. 27

    Scheller, J., Guhrs, K. H., Grosse, F. & Conrad, U. Production of spider silk proteins in tobacco and potato. Nature Biotechnol. 19, 573–577 (2001).

    CAS  Google Scholar 

  28. 28

    O'Dell, J. T., Nagy, F. & Chua, N. H. Identification of DNA sequences required for activity of the cauliflower mosaic virus 35S promoter. Nature 313, 810–812 (1985).

    CAS  Google Scholar 

  29. 29

    Lawton, M. A. et al. Expression of a soybean β-conclycinin gene under the control of the cauliflower mosaic virus 35S and 19S promoters in transformed petunia tissues. Plant Mol. Biol. 9, 315–324 (1987).

    CAS  PubMed  Google Scholar 

  30. 30

    Kay, R., Chan, A., Daly, M. & McPherson, J. Duplication of CaMV-35S promoter sequences creates a strong enhancer for plant genes. Science 236, 1299–1302 (1987).

    CAS  PubMed  Google Scholar 

  31. 31

    Christensen, A. H. & Quail, P. H. Ubiquitin promoter-based vectors for high-level expression of selectable and/or screenable marker genes in monocotyledonous plants. Transgenic Res. 5, 213–218 (1996).

    CAS  Google Scholar 

  32. 32

    Vain, P., Finer, K. R., Engler, D. E., Pratt, R. C. & Finer, J. J. Intron-mediated enhancement of gene expression in maize (Zea mays L.) and bluegrass (Poa pratensis L.). Plant Cell Rep. 15, 489–494 (1996).

    CAS  PubMed  Google Scholar 

  33. 33

    Stoger, E. et al. Cereal crops as viable production and storage systems for pharmaceutical scFv antibodies. Plant Mol. Biol. 42, 583–590 (2000).

    CAS  PubMed  Google Scholar 

  34. 34

    Artsaenko, O., Kettig, B., Fiedler, U., Conrad, U. & Düring K. Potato tubers as a biofactory for recombinant antibodies. Mol. Breeding 4, 313–319 (1998).

    CAS  Google Scholar 

  35. 35

    Padidam, M. Chemically regulated gene expression in plants. Curr. Opin. Plant Biol. 6, 169–177 (2003).

    CAS  PubMed  Google Scholar 

  36. 36

    Padidam, M., Gore, M., Lu, D. L. & Smirnova, O. Chemical-inducible, ecdysone receptor-based gene expression system for plants. Transgenic Res. 12, 101–109 (2003).

    CAS  PubMed  Google Scholar 

  37. 37

    Cramer, C. L., Boothe, J. G. & Oishi, K. K. Transgenic plants for therapeutic proteins: linking upstream and downstream technologies. Curr. Top. Microbiol. Immunol. 240, 95–118 (1999).

    CAS  PubMed  Google Scholar 

  38. 38

    Schillberg, S., Zimmermann, S., Voss, A. & Fischer, R. Apoplastic and cytosolic expression of full-size antibodies and antibody fragments in Nicotiana tabacum. Transgenic Res. 8, 255–263 (1999). This paper compares the stability of identical scFv antibodies that are targeted to different compartments, and shows that the secretory pathway is generally much more suitable for antibody accumulation than the cytosol.

    CAS  PubMed  Google Scholar 

  39. 39

    De Jaeger, G. et al. High-level accumulation of single-chain variable fragments in the cytosol of transgenic. Petunia hybrida. Eur. J. Biochem. 259, 1–10 (1998).

    Google Scholar 

  40. 40

    Schouten, A., Rossien, J., Bakker, J. & Schots, A. Formation of disulfide bridges by a single-chain Fv antibody in the reducing ectopic environment of the plant cytosol. J. Biol. Chem. 277, 19339–19345 (2002).

    CAS  PubMed  Google Scholar 

  41. 41

    Conrad, U. & Fiedler, U. 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 (1998).

    CAS  PubMed  Google Scholar 

  42. 42

    Plasterk, R. H. A. & Ketting, R. F. The silence of the genes. Curr. Opin. Genet. Dev. 10, 562–567 (2000).

    CAS  PubMed  Google Scholar 

  43. 43

    Anandalakshmi, R. et al. A viral suppressor of gene silencing in plants. Proc. Natl Acad. Sci. USA 95, 13079–13084 (1998).

    CAS  Google Scholar 

  44. 44

    Gelvin, S. B. Agrobacterium-mediated plant transformation: the biology behind the “gene-jockeying” tool. Microbiol. Mol. Biol. Rev. 67, 16–23 (2003).

    CAS  PubMed  PubMed Central  Google Scholar 

  45. 45

    Britt, A. B. & May, G. D. Re-engineering plant gene targeting. Trends Plant Sci. 8, 90–95 (2003).

    CAS  PubMed  PubMed Central  Google Scholar 

  46. 46

    Veluthambi, K., Gupta, A. K. & Sharma, A. The current status of plant transformation technologies. Curr. Sci. India 84, 368–380 (2003).

    CAS  Google Scholar 

  47. 47

    Christou, P. Transformation technology. Trends Plant Sci. 1, 423–431 (1996).

    Google Scholar 

  48. 48

    Twyman, R. M., Stoger, E., Kohli, A. & Christou, P. in Genetic Engineering: Principles and Practice Vol. 24 (ed. Setlow, J. K.) 107–136 (Kluwer-Plenum, New York, 2002).

    Google Scholar 

  49. 49

    Martineau, B., Voelker, T. A. & Sanders, V. A. On defining T-DNA. Plant Cell 6, 1032–1033 (1994).

    PubMed  PubMed Central  Google Scholar 

  50. 50

    Hanson, B. et al. A simple method to enrich an Agrobacterium-transformed population for plants containing only T-DNA sequences. Plant J. 19, 727–734 (1999).

    CAS  PubMed  Google Scholar 

  51. 51

    Fu, X. et al. Linear transgene constructs lacking vector backbone sequences generate low-copy-number transgenic plants with simple integration patterns. Transgenic Res. 9, 11–19 (2000).

    CAS  PubMed  Google Scholar 

  52. 52

    Lerouge, P., Bardor, M., Pagny, S., Gomord, V. & Faye, L. N-glycosylation of recombinant pharmaceutical glycoproteins produced in transgenic plants: towards humanisation of plant N-glycans. Curr. Pharmaceutical Biotech. 1, 347–354 (2000).

    CAS  Google Scholar 

  53. 53

    Bardor, M. et al. Immunoreactivity in mammals of two typical plant glyco-epitopes: core α(1,3)-fucose and core xylose. Glycobiology 13, 427–434 (2003).

    CAS  PubMed  Google Scholar 

  54. 54

    Chargelegue, D., Vine, N. D., van Dolleweerd, C. J., Drake, P. M. & Ma, J. K. A murine monoclonal antibody produced in transgenic plants with plant-specific glycans is not immunogenic in mice. Transgenic Res. 9, 187–194 (2000). The first published paper that discusses the immunogenicity of a plant-derived glycosylated recombinant protein.

    CAS  PubMed  Google Scholar 

  55. 55

    Warner, T. G. in Carbohydrates in Chemistry and Biology (eds Ernst, B., Hart, G. W. & Sanay, P.) 1043–1064 (Wiley, New York, 2000).

    Google Scholar 

  56. 56

    Blixt, O., Allin, K., Pereira, L., Datta, A. & Paulson, J. C. Efficient chemoenzymatic synthesis of O-linked sialyl oligosaccharides. J. Am. Chem. Soc. 124, 5739–5746 (2002).

    CAS  PubMed  Google Scholar 

  57. 57

    Bakker, H. et al. Galactose-extended glycans of antibodies produced by transgenic plants. Proc. Natl Acad. Sci. USA 98, 2899–2904 (2001). In this study, a transgenic tobacco line that expressed the heavy and light chains of a murine antibody was crossed with a line that expressed human β-1,4-galactosyltransferase. The progeny produced antibodies 30% of which had partially galactosylated N-glycans, which provided a useful approach for the 'humanization' of plant glycans.

    CAS  PubMed  Google Scholar 

  58. 58

    Raju, T. S., Briggs, J., Borge, S. M. & Jones, A. J. S. Species-specific variation in glycosylation of IgG: evidence for the species-specific sialylation and branch-specific galactosylation and importance for engineering recombinant glycoprotein therapeutics. Glycobiology 10, 477–486 (2000). In this study, cell-specific glycosylation of immunoglobulins was studied by mass spectrometry and capillary electrophoresis/laser-induced fluorescence in 13 different animal systems. The glycan patterns were found to be unique in different species, which indicated that some might be more suitable than others for the production of human therapeutic proteins. The same might apply to plant systems, which were not considered in this paper.

    CAS  PubMed  Google Scholar 

  59. 59

    Borisjuk, N. V. et al. Production of recombinant proteins in plant root exudates. Nature Biotechnol. 17, 466–469 (1999).

    CAS  Google Scholar 

  60. 60

    Komarnytsky, S., Borisjuk, N. V., Borisjuk, L. G., Alam, M. Z. & Raskin, I. Production of recombinant proteins in tobacco guttation fluid. Plant Physiol. 124, 927–933 (2000).

    CAS  PubMed  PubMed Central  Google Scholar 

  61. 61

    Drake, P. M. W. et al. Rhizosecretion of a monoclonal antibody protein complex from transgenic tobacco roots. Plant Mol. Biol. 52, 233–241 (2003).

    CAS  PubMed  Google Scholar 

  62. 62

    Maliga, P. Engineering the plastid genome of higher plants. Curr. Opin. Plant Biol. 5, 164–172 (2002).

    CAS  PubMed  Google Scholar 

  63. 63

    Daniell, H., Khan, M. S. & Allison, L. Milestones in chloroplast genetic engineering: an environmentally friendly era in biotechnology. Trends Plant Sci. 7, 84–91 (2002).

    CAS  PubMed  PubMed Central  Google Scholar 

  64. 64

    Tregoning, J. S. et al. Expression of tetanus toxin fragment C in tobacco chloroplasts. Nucleic Acids Res. 31, 1174–1179 (2003).

    CAS  PubMed  PubMed Central  Google Scholar 

  65. 65

    Daniell, H., Lee, S. B., Panchal, T. & Wiebe, P. O. Expression of the native cholera B toxin subunit gene and assembly as functional oligomers in transgenic tobacco chloroplasts. J. Mol. Biol. 311, 1001–1009 (2001).

    CAS  PubMed  PubMed Central  Google Scholar 

  66. 66

    Khan, M. S. & Maliga, P. Fluorescent antibiotic resistance marker for tracking plastid transformation in higher plants. Nature Biotechnol. 17, 910–915 (1999).

    CAS  Google Scholar 

  67. 67

    Fischer, R., Emans, N., Schuster, F., Hellwig, S. & Drossard, J. Towards molecular farming in the future: using plant-cell-suspension cultures as bioreactors. Biotechnol. Appl. Biochem. 30, 109–112 (1999).

    CAS  PubMed  Google Scholar 

  68. 68

    Stöger, E. et al. Practical considerations for pharmaceutical antibody production in different crop systems. Mol. Breeding 9, 149–158 (2002). This paper considers in detail the factors that should be evaluated when choosing a crop system for the production of pharmaceutical proteins. The same scFv is expressed in many species to compare intrinsic yields, and features such as storage, distribution and biosafety are discussed, as well as economic factors.

    Google Scholar 

  69. 69

    Witcher, D. et al. Commercial production of β-glucuronidase (GUS): a model system for the production of proteins in plants. Mol. Breeding 4, 301–312 (1998).

    CAS  Google Scholar 

  70. 70

    Hood, E. E., Woodard, S. L. & Horn, M. E. Monoclonal antibody manufacturing in transgenic plants myths and realities. Curr. Opin. Biotechnol. 13, 630–635 (2002).

    CAS  PubMed  Google Scholar 

  71. 71

    Hood, E. E. From green plants to industrial enzymes. Enzyme Microbial Technol. 30, 279–283 (2002).

    CAS  Google Scholar 

  72. 72

    Zeitlin, L. et al. A humanized monoclonal antibody produced in transgenic plants for immunoprotection of the vagina against genital herpes. Nature Biotechnol. 16, 1361–1364 (1998).

    CAS  Google Scholar 

  73. 73

    Khoudi, H. et al. Production of a diagnostic monoclonal antibody in perennial alfalfa plants. Biotechnol. Bioeng. 64, 135–143 (1999).

    CAS  PubMed  Google Scholar 

  74. 74

    Perrin, Y. et al. Transgenic pea seeds as bioreactors for the production of a single-chain Fv fragment (scFV) antibody used in cancer diagnosis and therapy. Mol. Breeding 6, 345–352 (2000).

    CAS  Google Scholar 

  75. 75

    De Wilde, C., Peeters, K., Jacobs, A., Peck, I. & Depicker, A. Expression of antibodies and Fab fragments in transgenic potato plants: a case study for bulk production in crop plants. Mol. Breeding 9, 2871–282 (2002).

    Google Scholar 

  76. 76

    Schunmann, P. H. D., Coia, G. & Waterhouse, P. M. Biopharming the Simpli-RED™ HIV diagnostic reagent in barley, potato and tobacco. Mol. Breeding 9, 113–121 (2002).

    Google Scholar 

  77. 77

    McGarvey, P. B. et al. Expression of the rabies virus glycoprotein in transgenic tomatoes. Biotechnology 13, 1484–1487 (1995).

    CAS  PubMed  Google Scholar 

  78. 78

    Sala, F. et al. Vaccine antigen production in transgenic plants: strategies, gene constructs and perspectives. Vaccine 21, 803–808 (2003).

    CAS  PubMed  Google Scholar 

  79. 79

    Commandeur, U., Twyman, R. M. & Fischer, R. The biosafety of molecular farming in plants. AgBiotechNet 5, ABN 110 (2003).

    Google Scholar 

  80. 80

    Hare, P. D. & Chua, N. -H. Excision of selectable marker genes from transgenic plants. Nature Biotechnol. 20, 575–579 (2002).

    CAS  Google Scholar 

  81. 81

    Zuo, J. R., Niu, Q. W., Ikeda, Y. & Chua, N. H. Marker-free transformation: increasing transformation frequency by the use of regeneration-promoting genes. Curr. Opin. Biotechnol. 13, 173–180 (2002).

    CAS  PubMed  Google Scholar 

  82. 82

    Eastham, K. & Sweet, J. Genetically Modified Organisms (GMOs): the Significance of Gene Flow through Pollen Transfer. Environment Issue Report No. 28 (European Environment Agency, Copenhagen, 2002)

    Google Scholar 

  83. 83

    Kay, E., Vogel, T. M., Bertolla, F., Nalin, R. & Simonet, P. In situ transfer of antibiotic resistance genes from transgenic (transplastomic) tobacco plants to bacteria. Appl. Environ. Microbiol. 68, 3345–3351 (2002).

    CAS  PubMed  PubMed Central  Google Scholar 

  84. 84

    Smalla, K. et al. in Proceedings of the 6th International Symposium on the Biosafety of Genetically Modified Organisms 146–154 (Univ. Extension Press, Univ. of Saskatchewan, Canada, 2000).

    Google Scholar 

  85. 85

    Smyth, S. & Phillips, P. W. B. Product differentiation alternatives: identity preservation, segregation and traceability. AgBioForum 5, 30–42 (2002).

    Google Scholar 

  86. 86

    Schillberg, S., Fischer, R. & Emans, N. Molecular farming of recombinant antibodies in plants. Cell. Mol. Life Sci. 60, 433–445 (2003). This is a comprehensive discussion of the technical issues concerning the production of antibodies in plants, which is treated in much more detail than is possible in the present review.

    CAS  PubMed  Google Scholar 

  87. 87

    Stoger, E., Sack, M., Fischer, R. & Christou, P. Plantibodies: applications, advantages and bottlenecks. Curr. Opin. Biotechnol. 13, 161–166 (2002).

    CAS  PubMed  Google Scholar 

  88. 88

    McCormick, A. A. et al. 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 (1999).

    CAS  PubMed  Google Scholar 

  89. 89

    Larrick, J. W., Yu, L., Naftzger, C., Jaiswal, S. & Wycoff, K. Production of secretory IgA antibodies in plants. Biomolecular Eng. 18, 87–94 (2001). This paper presents a useful summary of recent advances in the plant-based production of secretory IgAs with a discussion of purification methods and production costs.

    CAS  Google Scholar 

  90. 90

    Ma, J. K. et al. Characterization of a recombinant plant monoclonal secretory antibody and preventive immunotherapy in humans. Nature Med. 4, 601–606 (1998).

    CAS  PubMed  Google Scholar 

  91. 91

    Vaquero, C. et al. A carcinoembryonic antigen-specific diabody produced in tobacco. FASEB J. 16, 408–410 (2002).

    CAS  PubMed  Google Scholar 

  92. 92

    Kathuria, S. et al. Efficacy of plant-produced recombinant antibodies against HCG. Human Reproduction 17, 2054–2061 (2002).

    CAS  PubMed  Google Scholar 

  93. 93

    Miele, L. Plants as bioreactors for pharmaceuticals: regulatory considerations. Trends Biotechnol. 15, 45–50 (1997).

    CAS  PubMed  Google Scholar 

  94. 94

    Emlay, D. in Plants as Factories for Protein Production (eds Hood, E. E. & Howard, J.) 175–180 (Kluwer Academic, New York, 2002).

    Google Scholar 

  95. 95

    Lloyd-Evans, M. & Nair, A. in Biopharming: the Emerging World Market of Plant-based Therapeutics. Theta Report No. 1214 63–80 (PJB Medical Publications Inc., New York, 2002).

    Google Scholar 

  96. 96

    Stoger, E., Schillberg, S., Twyman, R. M., Fischer, R. & Christou, P. in Methods in Molecular Biology. Antibody Engineering: Protocols and Methods 2nd edn (ed. Lo, B. K. C.) (Humana Press Inc., New Jersey, in the press).

  97. 97

    Schillberg, S., Zimmermann, S., Findlay, K. & Fischer, R. Plasma membrane display of anti-viral single chain Fv fragments confers resistance to tobacco mosaic virus. Mol. Breeding 6, 317–326 (2000).

    CAS  Google Scholar 

  98. 98

    Sijmons, P. C. et al. Production of correctly processed human serum albumin in transgenic plants. Biotechnology 8, 217–221 (1990).

    CAS  PubMed  Google Scholar 

  99. 99

    Zhu, Z. et al. Expression of human α-interferon in plants. Virology 172, 213–222 (1994).

    Google Scholar 

  100. 100

    Matsumoto, S., Ikura, K., Ueda, M. & Sasaki, R. Characterisation of a human glycoprotein (erythropoetin) produced in cultured tobacco cells. Plant Mol. Biol. 27, 1163–1172 (1995).

    CAS  PubMed  Google Scholar 

  101. 101

    Delaney, D. et al. in Plant Biotechnology: 2002 and Beyond. Proceedings of the 10th IAPTC&B Congress, Orlando, Florida. (ed. Vasil, I.) 393–394 (Kluwer Academic, Dordrecht, The Netherlands, 2002).

    Google Scholar 

  102. 102

    Torres, E. et al. Rice cell culture as an alternative production system for functional diagnostic and therapeutic antibodies. Transgenic Res. 8, 441–449 (1999).

    CAS  PubMed  Google Scholar 

  103. 103

    Terashima, M. et al. Production of functional human α1–antitrypsin by plant cell culture. Appl. Microbiol. Biotechnol. 52, 516–523 (1999).

    CAS  PubMed  Google Scholar 

  104. 104

    Düring, K., Hippe, S., Kreuzaler, F. & Schell, J. Synthesis and self-assembly of a functional monoclonal antibody in transgenic Nicotiana tabacum. Plant Mol. Biol. 15, 281–293 (1990).

    PubMed  Google Scholar 

  105. 105

    Ma, J. K.-C. et al. Generation and assembly of secretory antibodies in plants. Science 268, 716–719 (1995).

    CAS  PubMed  Google Scholar 

  106. 106

    Francisco, J. A. et al. Expression and characterization of bryodin 1 and a bryodin 1-based single-chain immunotoxin from tobacco cell culture. Bioconjug. Chem. 8, 708–713 (1997).

    CAS  PubMed  Google Scholar 

  107. 107

    Mayfield, S. P. et al. Expression and assembly of a fully active antibody in algae. Proc. Natl Acad. Sci. USA 100, 438–442 (2003).

    CAS  PubMed  Google Scholar 

  108. 108

    Streatfield, S. J. et al. Plant-based vaccines: unique advantages. Vaccine 19, 2742–2748 (2001).

    CAS  PubMed  Google Scholar 

  109. 109

    Ma, S. W. et al. Transgenic plants expressing autoantigens fed to mice to induce oral immune tolerance. Nature Med. 3, 793–796 (1997).

    CAS  PubMed  Google Scholar 

  110. 110

    Yu, J. & Langridge, W. H. A plant-based multicomponent vaccine protects mice from enteric diseases. Nature Biotechnol. 19, 548–552 (2001).

    CAS  Google Scholar 

  111. 111

    Lamphear, B. J. et al. Delivery of subunit vaccines in maize seed. J. Control Release 83, 169–180 (2002).

    Google Scholar 

Download references


The authors are grateful to R. Twyman for critical assessment and help with manuscript preparation.

Author information



Corresponding author

Correspondence to Julian K-C. Ma.

Related links

Related links




Further information

Epicyte Pharmaceutical

Large Scale Biology Corp.




Pew Initiative on Food and Biotechnology

Phytomedics, Inc.


SemBioSys Genetics Inc.

Sigma Inc.



(scFvs). Monoclonal antibody derivatives that comprise a single polypeptide in which the variable regions of the heavy and light immunoglobulin chains are joined together by a flexible linker. scFvs are advantageous because only one transgene is required, and the molecules themselves are small and lack the effector functions of normal antibodies; however, a disadvantage is that they are univalent, whereas serum antibodies are divalent.


The large-scale production of recombinant proteins in living cells or organisms; frequently applied to the use of crop plants or domestic animals as expression hosts because of the allusion to agriculture.


In the context of this article, a gene or protein that is not derived from the species in which it is expressed.


A transgenic plant in which the transgene is found in the plastid genome rather than the nuclear genome.


A recombinant antibody that comprises the heavy- and light-chain variable regions joined by a flexible peptide linker. The linker is long enough to allow separation of the domains so that two of the polypeptides can assemble into a dimer, making the antibody divalent.


A recombinant antibody in which the heavy- and light-chain variable regions are part of the same polypeptide chain, which also includes the heavy-chain hinge region and one heavy-chain constant domain.


Usually leaves of tobacco (although many other species can be used) that are transiently transformed with Agrobacterium tumefaciens, which results in the transient expression of recombinant proteins. This is a useful strategy for testing expression constructs and obtaining small amounts of protein for analysis before going to the expense of transgenics.


Producing toxins in the gut that specifically affect the intestinal mucosa.


(Dicots). Broad-leaf flowering plants the seeds of which contain two cotyledons (embryonic seed leaves that either remain in the seed when the plant germinates or emerge and become green). Examples include potato, tomato, tobacco and all peas and beans.


Narrow-leaf plants the seeds of which contain one cotyledon. Examples include cereals, grasses, orchids and lilies.


A single antigenic determinant on a protein that is recognized by an antibody. A single protein can have many epitopes.


Interstitial cells in the testis that are responsible for the production of male sex hormones, such as testosterone, and are important in male sexual differentiation.


An in vitro mutagenesis procedure that is often carried out using the polymerase chain reaction in which specific mutations are introduced into a DNA molecule.


A short sequence of mainly hydrophobic amino acids at the N-terminus of secreted proteins. This peptide is captured by a signal-recognition particle as it emerges from the ribosome, which allows the ribosome to be transported to the endoplasmic reticulum.


Proteins the function of which is to ensure correct folding of other proteins during or after synthesis, or the refolding of denatured proteins.


The extracellular space. In plants, this is a large and continuous network of cavities under the cell wall. Proteins that are secreted from the cell often remain trapped here.


When transgenes integrate into genomic DNA, the expression level is often influenced by the surrounding chromatin. Local regulatory elements, such as enhancers, also influence transgene expression. Position effects lead to wide variations in transgene expression levels, even in plants that are transformed with identical constructs.


Short peptide sequences added to recombinant proteins, which bind strongly to particular affinity matrices and can be used to purify recombinant proteins.


Transformation that is achieved using the natural gene-transfer mechanism of Agrobacterium tumefaciens.


Transformation that is achieved by mixing walled plant cells with silicon carbide fibres that penetrate the cell wall and membrane, which generate pores through which DNA can be taken up into the cell.


Transformation that is achieved by exposing cells or protoplasts to a brief pulse of electricity, which results in the formation of transient membrane pores through which DNA can be taken up into the cell.


Any technique for introducing DNA into unwalled plant cells (protoplasts), such as calcium phosphate transfection, PEG transfection or electroporation.


Imperfect 25 bp direct repeat sequences that flank the piece of DNA that is transferred to the plant genome by Agrobacterium tumefaciens. These sequences are recognized by the bacterial VIRD1 and VIRD2 proteins, which form an endonuclease complex. Cleavage of the border sequences initiates T-DNA transfer.


A hybrid cell line that is created by fusing a mortal antibody-producing B-lymphocyte with an immortalized myeloma line. The hybridoma line is immortal and produces a continuous supply of a particular monoclonal antibody.


A family of flowering plants (order Solanales) that comprise 100 genera and 2,500 species, many of which are economically important as food or medicinal crops. Examples include tobacco, potato and tomato.


Callus tissue is undifferentiated plant tissue, which grows when seeds or explants are cultured on media that contains an appropriate balance of plant hormones. Friable callus tissue is easily broken into fragments.


Batch fermentation is a closed system in which all of the substrate is added at the beginning, whereas in the fed-batch process the substrate is added in increments as fermentation proceeds. Continuous fermentation is an open system in which substrate is added continuously at a steady rate. Perfusion fermentation is a continuous process that allows cells to be grown at high density, and so results in increased biomass and product yields.


The soil zone that surrounds plant roots, which is rich in microorganisms and in which interactions occur between plants and microbes.


Fluid that seeps from the apoplast onto the leaf surface. In plants with large leaves, such as tobacco, large amounts of guttation fluid can be produced each day.

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Ma, JC., Drake, P. & Christou, P. The production of recombinant pharmaceutical proteins in plants. Nat Rev Genet 4, 794–805 (2003).

Download citation

Further reading


Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing