Application of transgenic plants as factories for producing biopharmaceuticals

Paper Details

Review Paper 01/04/2014
Views (187) Download (6)

Application of transgenic plants as factories for producing biopharmaceuticals

Hadi Hashemzade, Alireza Zebarjadi, Nasim Akhshi, Kazem Aghaee
J. Bio. Env. Sci.4( 4), 58-74, April 2014.
Certificate: JBES 2014 [Generate Certificate]


Antibodies (Ab) are glycoprotein compounds produced by the vertebrate immune-system. They recognize and bind to their aim antigens with high affinity and specificity, which enable them to be used for numerous applications, such as the diagnosis, prevention, resistance to pathogens of plant, and cure of human and animal illness. The importance of antibodies as an in vitro investigation tool has been incorporate in vivo applications in basic researches of proteins and other compounds. Recently, the production of antibodies in plants because many reasons, including low-cost production, ease to storage large-scale production, safety and etc. is proposed. This technology is referring to the plantibody. Plantibody approach is one of the new events in molecular technology field. Plants as a bio-factory can be also used for biopharmaceutical and drugs production. Pharmaceuticals that derived from plants are poised to become the next major commercial development in plant biotechnology. But, however, there are some concerns in this context. In the present article, we try to summarize the recent advances made in the use of transgenic plants as biological factories for the production of drugs, vaccine, and biopharmaceuticals also discuss the plantibodies as a low-cost, efficient method for the production of biological materials with demonstrated utility to the pharmaceutical industry or medical community. In addition, a referring to applications of antibodies produced in plants and has been compared with other expression system.


Agrios J. 1997. Plant pathology. Academic press 365P.

Aliahmadi A, Rahmani N, Abdollahi M. 2006. Plant-derived human vaccines; An overview. Int. J. Pharmacol 2: 268-279.

Andersen D, Krummen CL. 2002. Recombinant protein expression for therapeutic applications. Curr Opin Biotechnol 13: 117–123.

Arakawa T, Chong DKX, Merrit JL, Langridge WHR. 1997. Expression of cholera toxin B subunit oligomers in transgenic potato plants. Transgenic Res 6: 403–413.

Artsaenko O, Kettig B, Fiedler U, Conrad U, During K. 1998. Potato tubers as a biofactory for recombinant antibodies. Mol Breed 4: 313–319.

Ata Saei A, Ghanbari P, Barzegar A. 2012. Haematococcus as a promising cell factory to produce recombinant pharmaceutical proteins. Mol Biol Rep DOI 10.1007/s11033,012,1861.

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-apreclinical and phase I investigation. PLoS One 43: e4792.

Azhdari H. 2009. Cloning and transformation of human IFN gene to tobacco plants. M.Sc. thesis. Department of Biotechnology, Faculty of Agriculture, Tarbiat Modarres University. Tehran – Iran.

Bagheri KH. 2009. Gamma,Oleosin interferon gene transfer to Canola and study of transgenic plants. PhD Thesis Department of Plant Breeding Faculty of Agriculture Tarbiat Modarres University Tehran – Iran.

Barta A, Sommergruber K, Thompson D, Hartmuth K, Matzke M, Matzke A. 1986. The expression of a nopaline synthase human growth hormone chimaeric gene in transformed tobacco and sunflower callus tissue. Plant Mol Biol 6: 347–57.

Beaujean A, Ducrocq,Assaf C, Sangwan RS, Lilius G, Bulow L. 2000. Sangwan, Norreel, B,S, Engineering direct fructose production in processed potato tubers by expressing a bifunctional alpha, amylase/glucose isomerase gene complex. Biotech Bioeng 70: 9–16.

Biemelt S, Sonnewald U, Galmbacher P, Willmitzer L, Müller M. 2003. Production of human papillomavirus type 16 virus,like particles in transgenic plants. J. Virol 7717: 9211–9220.

Bornke F, Hajirezaei M, Sonnewald U. 2002. Potato tubers as bioreactors for palatinose production. Biotech 96: 119–124.

Cabanes,Macheteau M, Fitchette,Lainé AC, Loutelier,Bourhis C, Lange C, Vine ND, Ma JK, Lerouge P, Faye L. 1999. N,Glycosylation of a mouse IgG expressed in transgenic tobacco plants. Glycobiology 9: 65-72.

Cardoso RM, 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.

Chen YD, Chen TA. 1998. Expression of engineered antibodies in plants, a possible tool for spiroplasma and phytoplasma disease control. Phytopathology 88: 1367–1371.

Chintalacharuvu KR, Morrison SL. 1999. Production and characterization of recombinant IgA. Immunotechnology 4: 65-74.

Cox KM, Sterling JD, Regan JT, GasdaskaJR, 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. Nature Biotechnology 24: 1591-1597.

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.

Crofls AJ, Denecke J. 1998. Calreticulin and canexin in plants. Tends plant Sci 3: 396-399.

D’Arcy CJ, Torrance L, Matrin RR. 1989. Discrimination among luteoviruses and their strains by monoclonal antibodies and identification of common epitopes. Phytopathology 79: 869–873.

Dalal M, Dani RG, Kumar PA. 2006. Current trends in the genetic engineering of vegetable crops. Sci. Hortic 107: 215-225.

Danielle H, Khan MS, Allison L. 2002. Milestones in chloroplast genetic engineering, an environmental friendly era in biotechnology. Trends Plant Sci 7: 84–91.

Danielle H, Stephen JS, Wycoff K. 2001. Medical molecular farming, production of antibodies, biopharmaceuticals and edible vaccines in plants. trends in Plant Sci 6: 1360-1385.

De Neve M, de Loose M, Jacobs A, van Houdt H, Kaluza B Weidle U, Depicker A. 1993. Assembly of an antibody and its derived antibody fragment in Nicotiana and Arabidopsis. Transgenic Res 2: 27–37.

Desai P, Shrivastava N, Padh H. 2010. Production of heterologous protein in plants, strategies for optimal expression. BiotechnolAdv 28: 427–435.

Drake PM, Chargelegue DM, Vine ND, van Dolleweerd CJ, Obregon P, Ma JK. 2003. Rhizosecretion of a monoclonal antibody protein complex from transgenic tobacco roots. Plant Mol Biol 52: 33-41.

Düring K, Hippe S, Kreuzaler F, Schell J. 1990. Synthesis and selfassembly of a functional monoclonal antibody in transgenic Nicotiana tabacum. Plant Mol Biol 15: 281-93.

Ehsani P, Khabiri A, Domansky NN. 1997. Polypeptides of hepatitis B surface antigen produced in transgenic potato. Gene 190: 107–1.1

Erokhina TN, Kastalieva TB, Mozhaeva KA. 2002. Preparing of monoclonal antibodies against BYDV using viruses purified from naturally infected plants, Proceedings of International Symposium. “Barley Yellow Dwarf Disease, Recent Advances and Future Strategies” 1–5 Sept El Batan Texcoco Mexico. 101–103.

Erokhina TN. 1995. Monoclonal antibodies to barley yellowdwarf virus, An immune enzyme test system for virus diagnostics. Bioorg Khim in Russian. 21: 256–260.

Fiedler U, Conrad U. 1995. High-level production and long-term storage of engineered antibodies in transgenic tobacco seeds. Biotech 1310: 1090-1093.

Fischer R, Richard M, Schillberg S. 2003. Production of antibodies in plants and their use for global health. Vaccine 21: 820–825.

Fischer R, Twyman MR, Hellwig S, Drossard J, Schillberg S. 2006. Facing the Future with Pharmaceuticals from Plants. Biotechnology and Sustainable Agriculture and Beyond, Proceedings of the 11th IAPTC&B Congress.

Floss DM, Falkenburg D, Conrad U. 2007. Production of vaccines and therapeutic antibodies for veterinary applications in transgenic plants. an overview Transgenic Res 163: 315,332.

Gething MJ, Sambrook J. 1992. Protein folding in the cell, Nature 355: 33-45.

Ghasempour HR, Kahrizi D, Mehdiah N. 2007. New Discussions in Biotechnology. Razi University Press 274p.

Giddings G, Allison G, Brooks D, Carter A. 2000. transgenic plants as factories for biopharmaceuticals. Nature America Inc 1811: 1151-5.

Haq TA, Mason HS, Clements JD, Arntzen CJ. 1995. Oral immunization with a recombinant bacterial antigen produced in transgenic plants. Science 268: 714-716.

He Y, Xu J, Tang C, Wu Y. 2005. Facilitation of pentachlorophenol degradation in the rhizosphere of ryegrass Loliumperenne L. Soil Biol Biochem 37: 2017–2024.

Hiatt A, Cafferkey R, Bowdish K. 1989. Production of antibodies in transgenic plants. Nature 3426245: 76-78.

Honari H. 2008. Expression of PA gene from Bacillus antracis in Iranian lettuce Lactuca sativa. Ph.D. thesis, Department of Plant Breeding, College of Agriculture, Tehran University. Tehran, Iran.

Hood EE, Witcher DR, Maddock S, Meyer T, Baszczynski C, Bailey M, Flynn P, Register J, Marshall L, Bond D, Kulisek E, Kusnadi AR, Evangelista R, Nikolov ZL, Wooge C, Mehigh RJ, Hernan R, Kappel WK, Ritland D, Li CP, Howard JA. 1997. Commercial production of avidin from transgenic maize, characterization of transformant, production, processing, extraction and purification. Mol Breeding 3: 291-306.

Hood EE, Woodard SL, Horn ME. 2002. Monoclonal antibody manufacturing in transgenic plants, myths and realities. Curr Opin Biotechnol 13: 630, 635.

Xiao XW, Chu PWG, Frenkel MJ, Tabe LM, Shukla DD, Hanna PJ, Higgins TJV, Muller WJ, Ward CW. 2000. Antibody,mediated improved resistance to ClYVV and PVY infections in transgenic tobacco plants expressing a single,chain variable region antibody. Mol Breed 6, 421–431.

Xu J, Okada Sh, Tan L, Goodrum KJ, Kopchick JJ, Kieliszewski MJ. 2010. Human growth hormone expressed in tobacco cells as an arabinogalactan, protein fusion glycoprotein has a prolonged serum life, Transgenic Res 19: 849–867.

Ismaili A, Jalali Javaran M, Rasaee MJ, Rahbarizadeh F, RajabiMemari H. 2006. Cloning and expression of recombinant camelid single,domain antibody in Tobacco. Iranian Journal of Biotechnology 43: 162-168.

Jaeger GD, Wilde CD, Eeckhout D, Fiers D, Depicker A. 2000. The plantibody approach, expression of antibody genes in plants to modulate plant metabolism or to obtain pathogen resistance. Plant Mol Biol 43: 419–428.

Jalali Javaran M, Mohebodini M, Masoumi Asl A, Saifi Nabi Abad H, Alizadeh H, Mahbodi F, Ismail A, Rajabi Memari H, Moini A, Honari H, Bagheri Kh, Yaghobi MM, Zebarjadi AR, Rasai MJ, Shakib AM, Rahbarizadeh F, Masoumi H, Forozandeh Moghadam M, Sharifi, Sirchi GR, Dymyad S, Sadat Noori SA, Vishlaghi N, Hosseini Pour A, Taheri Javan N, Razmi Sh, Rahimi Far P, Latif B, Abdolinasab M, Azhdari H, Poorkhaleghi M, Razmi A, Khosravi H, Kazemi H. 2009. The success of molecular farming in Iran. J. Agri Biotech 11: 19-48.

Jobling SA, Cees AM, Van Der Honde P, Punt J. 2003. Immunomodulation of enzyme function in plants by single domain antibody fragments. Nat Biotechnol 21: 77–80.

Kaiser J. 2008. Is the drought over for pharming? Science 320: 473,475.

Kay E, Drake PMW, Christou P. 2002. In situ transfer of antibiotic resistance genes from transgenic transplastomic. tobacco plants to bacteria. Appl Environ Microb 68: 3345–3351.

Langridge WHR. 2000. Edible vaccines. Scientific American 2833: 66-71.

Larrick JW, Thomas DW. 2001. Producing proteins in transgenic plants and animals. Curr Opin Biotechnol 12: 411–418.

Larrick JW, Yu L, Chen J, Jaiswal S, Wycoff K. 1998. Production of antibodies in transgenic plants. Res Immunol 149: 603–608.

Larrick JW, Yu L, Naftzger C, Jaiswal S, Wycoff K. 2001. Production of secretory IgA antibodies in plants. Biomol Eng 18: 87–94.

Lauterslager TGM, Florack DEA, van derWalTJ, Molthoff JW, Langeveld JPM, Bosch D. 2001. Oral immunization of naive and primed animals with transgenic potato tubers expressing LT, B. Vaccine 19: 49–55.

Lee SM, Kang K, Chung H, Yoo SH, Xu XM, Lee SB, Cheong JJ, Danielle H, Kim M. 2006. Plastid transformation in the monocotyledonous cereal crop, rice (Oryza sativa) and transmission of transgenes to their progeny. Mol. Cells 21: 401−410.

Lentz EM, Segretin ME, Morgenfeld MM, Wirth SA, Dus Santos MJ, Mozgovoj MV, Wigdorovitz A, Bravo-Almonacid FF. 2010. High expression level of a foot and mouth disease virus epitope in tobacco transplastomic plants. Planta 231: 387–395.

Lico C, Desideio A, Banchieri S, Benvenetu E. 2005. Plants as bio factories, Production of pharmaceutical recombinant proteins. Proceedings of the International Congress “In the Wake of the Double Helix, From the Green Revolution to the Gene Revolution”, Bologna, Italy 577,593.

Ma JK, Drake PM, Christou P. 2003. The production of recombinant pharmaceutical proteins in plants. Nat Rev Genet 4: 794–805.

Ma JK, Hiatt A, Hein M, Vine ND, Wang F, Stabila P, van Dolleweerd C, Mostov K, Lehner T. 1995. Generation and assembly of secretory antibodies in plants. Science 268: 716 – 719.

Ma JKC, Hein M. 1995. Immunotherapeutic potential of antibodies produced in plants. Trends Biotechnol 13: 522–527.

Ma JKC, Hiatt A. 1996. Expressing antibodies in plants for immunotherapy. In, Owen MRL, Pen J eds. transgenic plants, a production of pharmaceutical proteins. John Wiley & Sons, Chichester, U.K. 229-243pp.

Mason HS, Lam DMK, Arntzen CJ. 1992. Expression of hepatitis B surface antigen in transgenic plants. Proc. Natl. Acad. Sci. USA. 89: 11745-11749.

Mayfield SP, Franklin SE, Lerner RA. 2003. Expression and assembly of a fully active antibody in algae. ProcNatlAcadSci USA 100: 438–442.

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. 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.

McGarvey PB, Hammond J, Dienelt MM, Hooper DC, Fu ZF, Dietzschold B, Koprowski H, Michaels FH. 1995. Expression of the rabies virus glycoprotein in transgenic tomatoes. Bio Technol 13: 1484-1487.

Misaki R, Kimura Y, Palacpac NQ, Yoshida S, Fujiyama K, Seki T. 2003. Plant cultured cells expressing human beta1, 4, galactosyltransferase secrete glycoproteins with galactose,extended N,linked glycans. Glycobiology 13: 199-205.

Mor TS, Sternfeld M, Soreq H, Arntzen CJ, Mason HS 2001. Expression of recombinant human acetylcholinesterase in transgenic tomato plants. Biotechnol Bioeng 75: 259-266.

Noris E, Poli A, Cojoca R, Ritta M, Cavallo F, Vaglio S, Matic S, Landolfo S. 2011. A human papillomavirus 8 E7 protein produced in plants is able to trigger the mouse immune system and delay the development of skin lesions. Arch Virol 156: 587–595.

Obembe OO. 2010. Plant Biotechnology on a flight, Is Africa on board?. Life Science Journal 74: 14-25.

Pen J, van Ooyen AJJ, van den Elzen PJM, Rietveld K, Hoekema A. 1992. Direct screening for high,level expression of an introduced α,amylase gene in plants. Plant Mol Biol 18: 1133,1139.

Pujol M, Ramírez NI, Ayala M, Gavilondo JV, Valdés R, Rodríguez M, Brito J, Padilla S, Gómez L, Reyes B, Peral R, Pérez M, Marcelo JL, Milá L, Sánchez RF, Páez R, Cremata JA, Enríquez G, Mendoza, Ortega MO, Borroto C. 2005. An integral approach towards a practical application for a plant, made monoclonal antibody in vaccine purification. Vaccine 23: 1833-1837.

Purwar Sh, Soma S. Marla, US, Kumar SA. 2009. Basal expression studies of cystatins during specific growth stages of wheat spikes for defining their possible role in differential and stage dependent immunity against Karnal bunt Tilletiaindica. Mol Biol Rep 37: 1377–1389.

Ramírez N, Rodríguez M, Ayala M, Cremata J, Pérez M, Martínez A, Linares M, Hevia Y, Páez R, Valdés R, Gavilondo JV, Selman,Housein G. 2003. Expression and characterization of an anti, hepatitis B surface antigen. glycosylated mouse antibody in transgenic tobacco Nicotiana tabacum. plants and its use in the immunopurification of its target antigen. Biotechnol Appl Bioc 383: 223-230.

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.

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.

Rogers HG, Parkers HC. 1995. transgenic plants and the environment. J. of Exp. Bot 286: 467-488.

Ruggiero F, Exposito JY, Bournat P, Gruber V, Perret S, Comte J, Olagnier B, Garrone R, Theisen M. 2000. Triple helix assembly and processing of human collagen produced in transgenic tobacco plants. FEBS Lett 469: 132-136.

Schillberg S, Emans N, Fischer R. 2002. Antibody molecular farming in plants and plant cells. Phytochem Rev 1: 45–54.

Schillberg S, Fischer R, Emans N. 2003. Molecular farming of recombinant antibodies in plants. Cell Mol Life Sci 60: 33–45.

Schillberg S, Twyman RM. 2007. Pharma-Planta, Recombinant pharmaceuticals from plants for human health. In, Engelhard M, Hagen K, Thiele F eds. Pharming, A New Branch of Biotechnology. European Academy 43: 13-30.

Schillberg S, Zimmermann S, Zhang MY, Fischer R. 2001. Antibody based resistance to plant pathogens. Transgenic Res 10: 1–12.

Sharma A, Sharma KMK. 2009. Plants as bioreactors, Recent developments and emerging opportunities. BIOTECHNOL ADV 27: 811-832.

Shin-Young H, Tae-Geum K, Tae-Ho K, Yong-Suk J, Moon-Sik Y. 2007. Production of an Anti-mouse Mhc Class I Monoclonal Antibody with Biological Activity in Transgenic Tobacco. Protein Expr. Purif 54: 134-8.

Sijmons P.C, Dekker BMM, Schrammeijer B, Verwoerd, TC, Van Den Elzen PJM, Hoekema A. 1990. Production of correctly processed human serum albumin in transgenic plants. Nat. Biotechnol 8: 217–221.

Smith ML, Mason HS, Shuler ML. 2002. Hepatitis B surface antigen HbsAg. expression in plant cell culture, kinetics of antigen accumulation in batch culture and its intracellular form. BiotechnolBioeng 80: 12–22.

Sojikul P, Buehner N, Mason HS. 2003. A plant signal peptide hepatitis B surface antigen fusion protein with enhanced stability and immunogenicity expressed in plant cells. Proc Natl Acad Sci USA 100: 2209–14.

Spoke A, Karner S. 2008. Plant molecular farming opportunities and challenges. European comission press 148p.

Staub JM, Garcia B, Graves J, Hajdukiewicz PT, Hunter P, Nehra N, Paradkar V, Schlittler M, Carroll JA, Spatola L, Ward D, Ye G and Russell DA. 2000. High-yield production of a human therapeutic protein in tobacco chloroplasts. Nat Biotechnol 18: 333-338.

Stiger M, Neuhaus G, Manma, T. 1991. Self assembly of immunoglobulins in the cytoplasm of the alga Acetabulavia meditennnce. Plant Sci 73: 181-190.

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

Stoger E, Sack M, Perrin Y, Vaquero C, Torres E, Twyman RM, Christou P, Fischer R 2002. Practical considerations for pharmaceutical antibody production in different crop systems. Mol Breeding 9: 149–158.

Strasser R, AltmannF, MachL, Glössl J, Steinkellner H. 2004. Generation of Arabidopsis thaliana plants with complex N-glycans lacking beta1-2-linked xylose and core alpha1-3-linked fucose. FEBS Lett 561: 132-136.

Streatfield SJ, Jilka JM, Hood EE, Turner DD, Bailey MR, Mayor JM, Woodard SL, Beifuss KK, Horn ME, Delaney DE, Tizard IR, Howard JA. 2001. Plant-based vaccines, unique advantages. Vaccine 19: 2742-2748.

Tackaberry ES, Dudani AK, Prior F, Tocchi M, Sardana R, Altosaar I, Ganz PR 1999. Development of biopharmaceuticals in plant expression systems, cloning, expression and immunological rectivity of human cytomegalovirus glycoprotein B UL55 in seeds of transgenic tobacco. Vaccine 17: 3020-3029.

Tacket CO, Mason HS, Losonsky, G, Clements JD, Levine MM, Arntzen CJ. 1998. Immunogenicity in humans of a recombinant bacterial antigen delivered in a transgenic potato. J Nat Med 4: 607-609.

Triguero A, Cabrera G, Cremata JA, Yuen CT, Wheeler J, Ramírez 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.

Tuboly T, Yu W, Bailey A, Degrandis S, Du S, Erickson L, Nagy E. 2000. Immunogenicity of porcine transmissible gastroenteritis virus spike protein expressed in plants. Vaccine 18: 2023-2028.

Voss A, Niersback M, Hain R, Hirsch HJ, Liao YC, Kreuzaler F, Fischer R. 1995. Reduced virus infectivity in N. tabacum secreting a TMVspecific full size antibody. Mol Breed 1: 39–50.

Walmsley  AM,  Arntzen  CJ.  2000.  Plants  for delivery of edible vaccines. Curr Opin Biotechnol 11: 126–129.

Wang HH, Yin WB, Hu ZM. 2009. Advances in chloroplast engineering. J. Genet. Genomics 36: 387−398.

Wang XG, Zhang GH, Liu ChX, Zhang YH, Xiao ChZ, Fang RX. 2001. Purified Cholera Toxin B Subunit from Transgenic Tobacco Plants Possesses Authentic Antigenicity. Biotech Bioeng 724: 490-494.

Woodard SL, Mayor JM, Bailey MR, Barker DK, Love RT, Lane JR, Delaney DE, McComas, Wagner JM, Mallubhotla HD, Hood EE, Dangott LJ, Tichy SE, JA Howard JA. 2003. Maize Zea mays.,derived bovine trypsin, characterization of the first large,scale, commercial protein product from transgenic plants. Biotechnol Appl Biochem 38: 123–130.

Yu J, Langridge WHR. 2001. A plant-based multicomponent vaccine protects mice from enteric diseases. Nat Biotechnol 19: 548 – 552.

Yuan Q, Hu W, Pestka JJ, He SY, Hart LP. 2000. Expression of a functional antizearalenone single-chain Fv antibody in transgenic Arabidopsis plants. Appl. Environ. Microbiol 66: 3499–3505.

Zhang X, Urry DW, Danielle H. 1996. Expression of an environmentally friendly synthetic protein,based polymer gene in transgenic tobacco plants. Plant Cell Rep 16: 174-179.

Zimmermann S, Schillberg S, Liao YC, Fischer R. 1998. Intracellular expression of TMV,specific single,chain Fv fragments leads to improved virus resistance in Nicotiana tabacum. Mol Breed 4: 369–379.