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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

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 lo...

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Autores principales: Song, Ilchan, Lee, Young Koung, Kim, Jin Wook, Lee, Seung-Won, Park, Se Ra, Lee, Hae Kyung, Oh, Soyeon, Ko, Kinarm, Kim, Mi Kyung, Park, Soon Ju, Kim, Dae Heon, Kim, Moon-Soo, Kim, Do Sun, Ko, Kisung
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Korean Society for Molecular and Cellular Biology 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8560589/
https://www.ncbi.nlm.nih.gov/pubmed/34711693
http://dx.doi.org/10.14348/molcells.2021.2002
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author Song, Ilchan
Lee, Young Koung
Kim, Jin Wook
Lee, Seung-Won
Park, Se Ra
Lee, Hae Kyung
Oh, Soyeon
Ko, Kinarm
Kim, Mi Kyung
Park, Soon Ju
Kim, Dae Heon
Kim, Moon-Soo
Kim, Do Sun
Ko, Kisung
author_facet Song, Ilchan
Lee, Young Koung
Kim, Jin Wook
Lee, Seung-Won
Park, Se Ra
Lee, Hae Kyung
Oh, Soyeon
Ko, Kinarm
Kim, Mi Kyung
Park, Soon Ju
Kim, Dae Heon
Kim, Moon-Soo
Kim, Do Sun
Ko, Kisung
author_sort Song, Ilchan
collection PubMed
description 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 T(1) transformants and obtained homozygous T(3) 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.
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spelling pubmed-85605892021-11-12 Effect of an Endoplasmic Reticulum Retention Signal Tagged to Human Anti-Rabies mAb SO57 on Its Expression in Arabidopsis and Plant Growth Song, Ilchan Lee, Young Koung Kim, Jin Wook Lee, Seung-Won Park, Se Ra Lee, Hae Kyung Oh, Soyeon Ko, Kinarm Kim, Mi Kyung Park, Soon Ju Kim, Dae Heon Kim, Moon-Soo Kim, Do Sun Ko, Kisung Mol Cells Research Article 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 T(1) transformants and obtained homozygous T(3) 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. Korean Society for Molecular and Cellular Biology 2021-10-31 2021-10-29 /pmc/articles/PMC8560589/ /pubmed/34711693 http://dx.doi.org/10.14348/molcells.2021.2002 Text en © The Korean Society for Molecular and Cellular Biology. All rights reserved. https://creativecommons.org/licenses/by-nc-sa/3.0/This is an open-access article distributed under the terms of the Creative Commons Attribution-NonCommercial-ShareAlike 3.0 Unported License. To view a copy of this license, visit http://creativecommons.org/licenses/by-nc-sa/3.0/ (https://creativecommons.org/licenses/by-nc-sa/3.0/)
spellingShingle Research Article
Song, Ilchan
Lee, Young Koung
Kim, Jin Wook
Lee, Seung-Won
Park, Se Ra
Lee, Hae Kyung
Oh, Soyeon
Ko, Kinarm
Kim, Mi Kyung
Park, Soon Ju
Kim, Dae Heon
Kim, Moon-Soo
Kim, Do Sun
Ko, Kisung
Effect of an Endoplasmic Reticulum Retention Signal Tagged to Human Anti-Rabies mAb SO57 on Its Expression in Arabidopsis and Plant Growth
title Effect of an Endoplasmic Reticulum Retention Signal Tagged to Human Anti-Rabies mAb SO57 on Its Expression in Arabidopsis and Plant Growth
title_full Effect of an Endoplasmic Reticulum Retention Signal Tagged to Human Anti-Rabies mAb SO57 on Its Expression in Arabidopsis and Plant Growth
title_fullStr Effect of an Endoplasmic Reticulum Retention Signal Tagged to Human Anti-Rabies mAb SO57 on Its Expression in Arabidopsis and Plant Growth
title_full_unstemmed Effect of an Endoplasmic Reticulum Retention Signal Tagged to Human Anti-Rabies mAb SO57 on Its Expression in Arabidopsis and Plant Growth
title_short Effect of an Endoplasmic Reticulum Retention Signal Tagged to Human Anti-Rabies mAb SO57 on Its Expression in Arabidopsis and Plant Growth
title_sort effect of an endoplasmic reticulum retention signal tagged to human anti-rabies mab so57 on its expression in arabidopsis and plant growth
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8560589/
https://www.ncbi.nlm.nih.gov/pubmed/34711693
http://dx.doi.org/10.14348/molcells.2021.2002
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