Horticulture, Environment, and Biotechnology : HEB

Korean Society of Horticultural Science (한국원예학회)
권호 표지
DOI QR코드

DOI QR Code

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

  • Song, Ilchan (Department of Medicine, College of Medicine, Chung-Ang University) ;
  • Park, Sol-Ah (Department of Medicine, College of Medicine, Chung-Ang University) ;
  • Han, Dalmuri (Division of Bacterial Disease Research, Korea Centers for Disease Control and Prevention) ;
  • Lee, Hae-Kyung (Division of Bacterial Disease Research, Korea Centers for Disease Control and Prevention) ;
  • An, Hyun Joo (Graduate School of Analytical Science and Technology, Chungnam National University) ;
  • Ko, Kisung (Department of Medicine, College of Medicine, Chung-Ang University)
  • Received : 2017.08.16
  • Accepted : 2017.08.29
  • Published : 2018.04.30
⟨ Previous Next ⟩

Abstract

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.

Keywords

Acknowledgement

Supported by : Chung-Ang University, Korean Rural Administration

References

  1. Carneiro JMT, Madrid KC, Maciel BCM, Arruda MAZ (2015) Arabidopsis thaliana and omics approaches: a review. J Integr OMICS. https://doi.org/10.5584/jiomics.v5i1.179
  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. https://doi.org/10.1007/s13337-012-0096-x
  3. Clough SJ, Bent AF (1998) Floral dip: a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana. Plant J 16:735-743. https://doi.org/10.1046/j.1365-313x.1998.00343 .x
  4. Gerngross TU (2004) Advances in the production of human therapeutic proteins in yeasts and filamentous fungi. Nat Biotechnol 22:1409-1414. https://doi.org/10.1038/nbt10 28
  5. Gomord V, Faye L (2004) Posttranslational modification of therapeutic proteins in plants. Curr Opin Plant Biol 7:171-181. https://doi.org/10.1016/j.pbi.2004.01.015
  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. https://doi.org/10.1007/Bf013 15415
  7. Hiatt A, Cafferkey R, Bowdish K (1989) Production of antibodies in transgenic plants. Nature 342:76-78. https://doi.org/10.1038/342076a0
  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. https://doi.org/10.1016/j.biotechadv.2009.07.004
  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. https://doi.org/10.1007/s13580-016-0045-5
  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. https://doi.org/10.3389/fpls.2017.00274
  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. https://doi.org/10.3389/fpls.2016.01037
  12. Ko K (2014) Expression of recombinant vaccines and antibodies in plants. Monoclon Antib Immunodiagn Immunother 33:192-198. https://doi.org/10.1089/mab.2014.0049
  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. https://doi.org/10.1073/pnas.08324 72100
  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):494-503
  15. Lee JH, Ko K (2017) Production of recombinant anti-cancer vaccines in plants. Biomol Ther 25:345-353. https://doi.org/10.4062/biomolther.2016.126
  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. https://doi.org/10.1371/journ al.pone.0068772
  17. Mason HS, Lam DMK, Arntzen CJ (1992) Expression of hepatitis-B surface-antigen in transgenic plants. PNAS 89:11745-11749. https://doi.org/10.1073/pnas.89.24.11745
  18. McGettigan JP (2010) Experimental rabies vaccines for humans. Expert Rev Vaccines 9:1177-1186. https://doi.org/10.1586/erv.10.105
  19. Moussavou G, Ko K, Lee JH, Choo YK (2015) Production of monoclonal antibodies in plants for cancer immunotherapy. Biomed Res Int. https://doi.org/10.1155/2015/30616 4
  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. https://doi.org/10.3389/fpls.2015.01040
  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. https://doi.org/10.1007/s1358 0-015-0097-y
  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-mannosetype N-glycans. Plant Biotechnol J 3:449-457. https://doi.org/10.1111/j.1467-7652.2005.00137.x
  23. Wurm FM (2004) Production of recombinant protein therapeutics in cultivated mammalian cells. Nat Biotechnol 22:1393-1398. https://doi.org/10.1038/nbt10 26
  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. https://doi.org/10.3390/ijms1 61226122

Cited by

  1. Expression and in vitro function of anti-cancer mAbs in transgenic Arabidopsis thaliana vol.53, pp.4, 2018, https://doi.org/10.5483/bmbrep.2020.53.4.106
  2. Effect of an Endoplasmic Reticulum Retention Signal Tagged to Human Anti-Rabies mAb SO57 on Its Expression in Arabidopsis and Plant Growth vol.44, pp.10, 2021, https://doi.org/10.14348/molcells.2021.2002