Horticulture, Environment, and Biotechnology

, Volume 59, Issue 2, pp 285–292 | Cite as

Expression, glycosylation, and function of an anti-rabies virus monoclonal antibody in tobacco and Arabidopsis plants

  • Ilchan Song
  • Sol-Ah Park
  • Dalmuri Han
  • Hae Kyung Lee
  • Hyun Joo An
  • Kisung KoEmail author
Research Report Tissue Culture/Biotechnology


Plants have emerged as one of the most attractive systems for producing human therapeutic proteins against viral diseases. These include diagnostic reagents, vaccines, and antibodies. This process is known as molecular biofarming. The objective of this study was to develop and evaluate tobacco and Arabidopsis as plant platforms for producing human anti-rabies monoclonal antibody (mAb). Both tobacco and Arabidopsis transgenic plants were generated by Agrobacterium-mediated transformation. Purification of mAb SO57K from each plant was performed with ammonium sulfate-mediated precipitation and protein A affinity columns. SDS–PAGE analysis showed that the purity of mAb SO57K obtained from each transgenic plant was similar, whereas Arabidopsis showed approximately twofold greater protein expression than tobacco. The N-glycosylation was not significantly different between proteins from the two plant species, with both showing oligo-mannose glycan structures. The mAbs SO57 derived from both the model plants had similar neutralizing efficacy against target virus strain CVS-11. Taken together, tobacco and Arabidopsis are both promising platforms for producing a human anti-rabies mAb.


Molecular biofarming Platform Tobacco Arabidopsis Transgenic plant Anti-rabies monoclonal antibody Neutralizing efficacy 



This research was supported by the Chung-Ang University Research Scholarship Grants in 2015 and the Korean Rural Administration (Grant Code # PJ011110).


  1. Carneiro JMT, Madrid KC, Maciel BCM, Arruda MAZ (2015) Arabidopsis thaliana and omics approaches: a review. J Integr OMICS. Google Scholar
  2. Chander V, Singh RP, Verma PC (2012) Development of monoclonal antibodies suitable for rabies virus antibody and antigen detection. Indian J Virol 23:317–325. CrossRefPubMedPubMedCentralGoogle Scholar
  3. Clough SJ, Bent AF (1998) Floral dip: a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana. Plant J 16:735–743. CrossRefPubMedGoogle Scholar
  4. Gerngross TU (2004) Advances in the production of human therapeutic proteins in yeasts and filamentous fungi. Nat Biotechnol 22:1409–1414. CrossRefPubMedGoogle Scholar
  5. Gomord V, Faye L (2004) Posttranslational modification of therapeutic proteins in plants. Curr Opin Plant Biol 7:171–181. CrossRefPubMedGoogle Scholar
  6. Goto H, Minamoto N, Ito H, Luo TR, Sugiyama M, Kinjo T, Kawai A (1995) Expression of the nucleoprotein of rabies virus in Escherichia-coli and mapping of antigenic sites. Arch Virol 140:1061–1074. CrossRefPubMedGoogle Scholar
  7. Hiatt A, Cafferkey R, Bowdish K (1989) Production of antibodies in transgenic plants. Nature 342:76–78. CrossRefPubMedGoogle Scholar
  8. Jamal A, Ko K, Kim HS, Choo YK, Joung H, Ko K (2009) Role of genetic factors and environmental conditions in recombinant protein production for molecular farming. Biotechnol Adv 27:914–923. CrossRefPubMedGoogle Scholar
  9. Kang Y, Shin YK, Park S-W, Ko K (2016) Effect of nitrogen deficiency on recombinant protein production and dimerization and growth in transgenic plants. Hortic Environ Biotechnol 57:299–307. CrossRefGoogle Scholar
  10. Kang YJ, Kim DS, Myung SC, Ko K (2017) Expression of a human prostatic acid phosphatase (PAP)-IgM Fc fusion protein in plants using in vitro tissue subculture. Front Plant Sci 8:274. PubMedPubMedCentralGoogle Scholar
  11. Kim DS, Song I, Kim J, Kim DS, Ko K (2016) Plant recycling for molecular biofarming to produce recombinant anti-cancer mAb. Front Plant Sci 7:1037. PubMedPubMedCentralGoogle Scholar
  12. Ko K (2014) Expression of recombinant vaccines and antibodies in plants. Monoclon Antib Immunodiagn Immunother 33:192–198. CrossRefPubMedPubMedCentralGoogle Scholar
  13. Ko KS, Tekoah Y, Rudd PM, Harvey DJ, Dwek RA, Spitsin S, Hanlon CA, Rupprecht C, Dietzschold B et al (2003) Function and glycosylation of plant-derived antiviral monoclonal antibody. PNAS 100:8013–8018. CrossRefPubMedPubMedCentralGoogle Scholar
  14. Ko KS, Ahn MH, Song MR, Choo YK, Kim HS, Ko KN, Jung HU (2008) Glyco-engineering of biotherapeutic proteins in plants. Mol Cells 25(4):494–503PubMedGoogle Scholar
  15. Lee JH, Ko K (2017) Production of recombinant anti-cancer vaccines in plants. Biomol Ther 25:345–353. CrossRefGoogle Scholar
  16. Lee JH, Park DY, Lee KJ, Kim YK, So YK, Ryu JS, Oh SH, Han YS, Ko K et al (2013) Intracellular reprogramming of expression, glycosylation, and function of a plant-derived antiviral therapeutic monoclonal antibody. PLoS ONE 8:e68772. CrossRefPubMedPubMedCentralGoogle Scholar
  17. Mason HS, Lam DMK, Arntzen CJ (1992) Expression of hepatitis-B surface-antigen in transgenic plants. PNAS 89:11745–11749. CrossRefPubMedPubMedCentralGoogle Scholar
  18. McGettigan JP (2010) Experimental rabies vaccines for humans. Expert Rev Vaccines 9:1177–1186. CrossRefPubMedPubMedCentralGoogle Scholar
  19. Moussavou G, Ko K, Lee JH, Choo YK (2015) Production of monoclonal antibodies in plants for cancer immunotherapy. Biomed Res Int. PubMedPubMedCentralGoogle Scholar
  20. Park SR, Lim CY, Kim DS, Ko K (2015) Optimization of ammonium sulfate concentration for purification of colorectal cancer vaccine candidate recombinant protein GA733-FcK isolated from plants. Front Plant Sci 6:1040. PubMedPubMedCentralGoogle Scholar
  21. Song I, Kim DS, Kim MK, Jamal A, Hwang K-A, Ko K (2015) Comparison of total soluble protein in various horticultural crops and evaluation of its quantification methods. Hortic Environ Biotechnol 56:123–129. CrossRefGoogle Scholar
  22. Triguero A, Cabrera G, Cremata JA, Yuen CT, Wheeler J, Ramirez 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. CrossRefPubMedGoogle Scholar
  23. Wurm FM (2004) Production of recombinant protein therapeutics in cultivated mammalian cells. Nat Biotechnol 22:1393–1398. CrossRefPubMedGoogle Scholar
  24. Yao J, Weng YQ, Dickey A, Wang KY (2015) Plants as factories for human pharmaceuticals: applications and challenges. Int J Mol Sci 16:28549–28565. CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

© Korean Society for Horticultural Science and Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  • Ilchan Song
    • 1
  • Sol-Ah Park
    • 1
  • Dalmuri Han
    • 2
  • Hae Kyung Lee
    • 2
  • Hyun Joo An
    • 3
  • Kisung Ko
    • 1
    Email author
  1. 1.Department of Medicine, College of MedicineChung-Ang UniversitySeoulKorea
  2. 2.Division of Bacterial Disease ResearchKorea Centers for Disease Control and PreventionOsongKorea
  3. 3.Graduate School of Analytical Science and TechnologyChungnam National UniversityDaejeonKorea

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