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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 Korean Society for Molecular and Cellular Biology 2021 Molecules and cells Vol.44 No.10
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<sub>1</sub> transformants and obtained homozygous T<sub>3</sub> 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.
Junsik Ahn,Ilchan Song,Dongjae Kim,Joon Chul Lee,Sungkwon Moon,Sooncheol Myoung,Kisung Ko 한국원예학회 2017 원예과학기술지 Vol.35 No.4
We performed seed germination tests to investigate the effects of seed sowing orientation on germination viability on peanut (Arachis hypogaea L.) sprouts. Specifically, we assessed the influence of seed sowing orientation on germination rate, seedling weight, and seedling length, as well as the seedling vigor index. The seeds were sown in oak tree sawdust at 3.0 cm depth. Four seed orientations were tested: vertical with the hypocotyl end down, vertical with the hypocotyl end up, horizontal with the hypocotyl end down, and horizontal with the hypocotyl end up. The mean seed germination percentages of the four seed orientations were significantly different (p < 0.01) and ranged from 25 to 91.7%. The vertical orientation with hypocotylend-down and hypocotyl-end-up orientations showed the highest (91.7%) and lowest (25%) germination rates, respectively. The vertical orientation with the hypocotyl end down produced the heaviest (4.9 g) seedlings and the longest hypocotyls (4.65 cm). This orientation also produced the longest true leaf + epycotyl (2.15 cm) and had the highest seedling vigor index (197.1). The seedlings had a straight growth pattern, whereas seedlings from seeds sown with the hypocotyl up had an awkward plumular hook shape. Taken together, to produce peanut sprouts, we recommend placing the seeds vertically with the hypocotyl end down because this orientation leads to a high germination rate, high biomass production, and high overall seedling quality.
Ju, Won Seok,Song, Ilchan,Park, Se-Ra,Seo, Sang Young,Cho, Jin Hyoung,Min, Sung-Hun,Kim, Dae-Heon,Kim, Ji-Su,Kim, Sun-Uk,Park, Soon Ju,Ko, Kisung,Choo, Young-Kug The Korean Society of Plant Biotechnology 2019 식물생명공학회지 Vol.46 No.3
Production of therapeutic monoclonal antibodies (mAbs) using a plant platform has been considered an alternative to the mammalian cell-based production system. A plant-derived mAb CO17-1AK ($mAb^P$ COK) can specifically bind to various types of cancer cell lines. The target protein of $mAb^P$ COK is the epithelial cell adhesion molecule (EpCAM) highly expressed in human epithelial cancer cells, including breast and colorectal cancer cells. It has been hypothesized that its overexpression supports tumor growth and metastasis. A ganglioside is extended well beyond the surfaces of the various cell membranes and has roles in cell growth, inflammation, differentiation, and carcinogenesis. However, the regulation of EpCAM gene expression in breast cancers and the role of gangliosides in oncogenesis are unclear. Here, the purpose of this study was to determine the effects of $mAb^P$ COK on human breast cancer cell proliferation, apoptosis, and ganglioside expression patterns. Our results show that treatment with $mAb^P$ COK suppressed the growth of breast cancer cells and induced apoptotic cell death. It also upregulated the expression of metastasis-related gangliosides in breast cancer cells. Thus, treatment with $mAb^P$ COK may have chemo-preventive therapeutic effects against human breast cancer.
Ahn, Junsik,Song, Ilchan,Kim, Dongjae,Lee, Joon Chul,Moon, Sungkwon,Myoung, Sooncheol,Ko, Kisung Korean Society of Horticultural Science 2017 원예과학기술지 Vol.35 No.4
We performed seed germination tests to investigate the effects of seed sowing orientation on germination viability on peanut (Arachis hypogaea L.) sprouts. Specifically, we assessed the influence of seed sowing orientation on germination rate, seedling weight, and seedling length, as well as the seedling vigor index. The seeds were sown in oak tree sawdust at 3.0 cm depth. Four seed orientations were tested: vertical with the hypocotyl end down, vertical with the hypocotyl end up, horizontal with the hypocotyl end down, and horizontal with the hypocotyl end up. The mean seed germination percentages of the four seed orientations were significantly different (p < 0.01) and ranged from 25 to 91.7%. The vertical orientation with hypocotyl-end-down and hypocotyl-end-up orientations showed the highest (91.7%) and lowest (25%) germination rates, respectively. The vertical orientation with the hypocotyl end down produced the heaviest (4.9 g) seedlings and the longest hypocotyls (4.65 cm). This orientation also produced the longest true leaf + epycotyl (2.15 cm) and had the highest seedling vigor index (197.1). The seedlings had a straight growth pattern, whereas seedlings from seeds sown with the hypocotyl up had an awkward plumular hook shape. Taken together, to produce peanut sprouts, we recommend placing the seeds vertically with the hypocotyl end down because this orientation leads to a high germination rate, high biomass production, and high overall seedling quality.
Won Seok Ju,Ilchan Song,Sung Youn Heo,Hyeon Gyu Lim,Sang Young Seo,Jin Hyoung Cho,Soon Ju Park,Young-Kug Choo 한국당과학회 2018 한국당과학회 학술대회 Vol.2018 No.07
An epithelial cell adhesion molecule (EpCAM) is a type I transmembrane protein expressed in the majority of normal epithelial tissues and overexpressed in the human epithelial cancers including breast cancer. Treatment with plant-derived anti-EpCAM mAb (mAbr COK) and Raw264. 7 cells inhibited cell growth in the human breast cancer cell line MCF -7. In MCF-7 treated with mAbr COK and Raw264.7 cells, expression of p53 and p21 increased, whereas the expression of G1 phase-related protein, cyclin D1, CDK4, cyclin E and CDK2 decreased. In addition of treatment with mAbP COK and Raw264. 7 cells decreased the expressiOn of anti-apoptotic protein Bcl-2, but the expression of pro-apoptotic proteins Bax, TNF-u, caspase-3, caspase-6 and caspase-9, increased. Moreover, cells treated with mAbr COK and Raw264. 7 cells expressed metastasis-related gangliosides GM1 and GD1a. These results suggest that mAbr COK has an effect on anti-cancer activity.
Won Seok Ju,Ilchan Song,Se-Ra Park,Sang Young Seo,Jin Hyoung Cho,Sung-Hun Min,Dae-Heon Kim,Ji-Su Kim,Sun-Uk Kim,Soon Ju Park,Kisung Ko,Young-Kug Choo 한국식물생명공학회 2019 JOURNAL OF PLANT BIOTECHNOLOGY Vol.46 No.3
Production of therapeutic monoclonal antibodies (mAbs) using a plant platform has been considered an alternative to the mammalian cell-based production system. A plant-derived mAb CO17-1AK (mAbP COK) can specifically bind to various types of cancer cell lines. The target protein of mAbP COK is the epithelial cell adhesion molecule (EpCAM) highly expressed in human epithelial cancer cells, including breast and colorectal cancer cells. It has been hypothesized that its overexpression supports tumor growth and metastasis. A ganglioside is extended well beyond the surfaces of the various cell membranes and has roles in cell growth, inflammation, differentiation, and carcinogenesis. However, the regulation of EpCAM gene expression in breast cancers and the role of gangliosides in oncogenesis are unclear. Here, the purpose of this study was to determine the effects of mAbP COK on human breast cancer cell proliferation, apoptosis, and ganglioside expression patterns. Our results show that treatment with mAbP COK suppressed the growth of breast cancer cells and induced apoptotic cell death. It also upregulated the expression of metastasis-related gangliosides in breast cancer cells. Thus, treatment with mAbP COK may have chemo-preventive therapeutic effects against human breast cancer.