Molecules and Cells

Korean Society for Molecular and Cellular Biology (한국분자세포생물학회)
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Effect of an Endoplasmic Reticulum Retention Signal Tagged to Human Anti-Rabies mAb SO57 on Its Expression in Arabidopsis and Plant Growth

  • Song, Ilchan (Department of Medicine, College of Medicine, Chung-Ang University) ;
  • Lee, Young Koung (Plasma Technology Research Center, National Fusion Research Institute) ;
  • Kim, Jin Wook (Department of Urology, College of Medicine, Chung-Ang University) ;
  • Lee, Seung-Won (Department of Medicine, College of Medicine, Chung-Ang University) ;
  • Park, Se Ra (Department of Medicine, College of Medicine, Chung-Ang University) ;
  • Lee, Hae Kyung (Division of Zoonotic and Vector Borne Diseases Research, Korea National Institute of Health) ;
  • Oh, Soyeon (Department of Medicine, College of Medicine, Chung-Ang University) ;
  • Ko, Kinarm (Department of Stem Cell Biology, Konkuk University School of Medicine) ;
  • Kim, Mi Kyung (Department of Pathology, College of Medicine, Chung-Ang University) ;
  • Park, Soon Ju (Division of Biological Sciences, Wonkwang University) ;
  • Kim, Dae Heon (Department of Biology, Sunchon National University) ;
  • Kim, Moon-Soo (Department of Chemistry, Western Kentucky University) ;
  • Kim, Do Sun (Vegetable Research Division, National Institute of Horticultural and Herbal Science, Rural Development Administration (RDA)) ;
  • Ko, Kisung (Department of Medicine, College of Medicine, Chung-Ang University)
  • Received : 2020.12.27
  • Accepted : 2021.08.27
  • Published : 2021.10.31
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Abstract

Transgenic Arabidopsis thaliana expressing an anti-rabies monoclonal antibody (mAb), SO57, was obtained using Agrobacterium-mediated floral dip transformation. The endoplasmic reticulum (ER) retention signal Lys-Asp-Glu-Leu (KDEL) was tagged to the C-terminus of the anti-rabies mAb heavy chain to localize the mAb to the ER and enhance its accumulation. When the inaccurately folded proteins accumulated in the ER exceed its storage capacity, it results in stress that can affect plant development and growth. We generated T1 transformants and obtained homozygous T3 seeds from transgenic Arabidopsis to investigate the effect of KDEL on plant growth. The germination rate did not significantly differ between plants expressing mAb SO57 without KDEL (SO plant) and mAb SO57 with KDEL (SOK plant). The primary roots of SOK agar media grown plants were slightly shorter than those of SO plants. Transcriptomic analysis showed that expression of all 11 ER stress-related genes were not significantly changed in SOK plants relative to SO plants. SOK plants showed approximately three-fold higher mAb expression levels than those of SO plants. Consequently, the purified mAb amount per unit of SOK plant biomass was approximately three times higher than that of SO plants. A neutralization assay revealed that both plants exhibited efficient rapid fluorescent focus inhibition test values against the rabies virus relative to commercially available human rabies immunoglobulins. KDEL did not upregulate ER stress-related genes; therefore, the enhanced production of the mAb did not affect plant growth. Thus, KDEL fusion is recommended for enhancing mAb production in plant systems.

Keywords

Acknowledgement

This research was supported by grants from Cooperative Research Program for Agriculture Science and Technology Development (project No. PJ016266), and Chung-Ang University Research Scholarship Grants in 2018.

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