caa a22 4500 605498482 CHVBK 20210128100546.0 cr unu---uuuuu 210128e20150801xx s 000 0 eng 10.1007/s00253-015-6601-6 doi (NATIONALLICENCE)springer-10.1007/s00253-015-6601-6 Expression of bioactive anti-CD20 antibody fragments and induction of ER stress response in Arabidopsis seeds [Elektronische Daten] [Dezhong Wang, Jisheng Ma, Difei Sun, Haiyan Li, Chao Jiang, Xiaokun Li] Seed-based expression system is an attractive platform for the production of recombinant proteins in molecular farming. Despite the many advantages of molecular farming, little is known about the effect of the different subcellular accumulation of recombinant proteins on the endoplasmic reticulum (ER) quality control system in host plants. In this study, we analyzed the expression of anti-CD20 antibody fragments in seeds of Arabidopsis thaliana (ecotype Columbia) and corresponding glycosylation mutants, and evaluated the influence of three different signal sequences on the expression levels of scFv-Fc of C2B8. The highest protein accumulation level, with a maximum of 6.12% total soluble proteins, was observed upon fusing proteins to the signal peptide of Arabidopsis seed storage albumin 2. The ER stress responses in developing seeds at 13days post-anthesis were also compared across different transgenic lines under normal and heat shock conditions. Based on the gene expression profiles of ER stress transducers, our results suggest that accumulation of antibody fragments in the ER exerts more stress on ER homeostasis. In addition, quantitative PCR results also implicate enhanced activation of ER-associated degradation in transgenic lines. Last but not the least, we also demonstrate the anti-tumor potency of plant-derived proteins by showing the anti-tumor activity of purified scFv-Fc proteins against Daudi cells. Together, our data implies that better understanding of the interaction between exogenous protein production and the cellular quality control system of the host plant is necessary for the development of an optimal expression strategy that will be especially beneficial to commercial protein manufacturing. Springer-Verlag Berlin Heidelberg, 2015 Arabidopsis seeds nationallicence Anti-CD20 antibody fragments nationallicence ADCC nationallicence CDC nationallicence ER stress response nationallicence ER-associated degradation nationallicence Wang Dezhong College of Life Sciences, Jilin University, 130021, Changchun, China aut Ma Jisheng School of Pharmaceutical Sciences, Zhejiang Key Laboratory of Biotechnology and Pharmaceutical Engineering, Wenzhou Medical University, Chashan College Park, 325035, Wenzhou, China aut Sun Difei School of Pharmaceutical Sciences, Zhejiang Key Laboratory of Biotechnology and Pharmaceutical Engineering, Wenzhou Medical University, Chashan College Park, 325035, Wenzhou, China aut Li Haiyan Ministry of Education Engineering Research Center of Bioreactor and Pharmaceutical Development, Jilin Agricultural University, Changchun, China aut Jiang Chao Ministry of Education Engineering Research Center of Bioreactor and Pharmaceutical Development, Jilin Agricultural University, Changchun, China aut Li Xiaokun College of Life Sciences, Jilin University, 130021, Changchun, China aut Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/16(2015-08-01), 6753-6764 0175-7598 99:16<6753 2015 99 253 https://doi.org/10.1007/s00253-015-6601-6 text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s00253-015-6601-6 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Wang Dezhong College of Life Sciences, Jilin University, 130021, Changchun, China aut NATIONALLICENCE 700 1- Ma Jisheng School of Pharmaceutical Sciences, Zhejiang Key Laboratory of Biotechnology and Pharmaceutical Engineering, Wenzhou Medical University, Chashan College Park, 325035, Wenzhou, China aut NATIONALLICENCE 700 1- Sun Difei School of Pharmaceutical Sciences, Zhejiang Key Laboratory of Biotechnology and Pharmaceutical Engineering, Wenzhou Medical University, Chashan College Park, 325035, Wenzhou, China aut NATIONALLICENCE 700 1- Li Haiyan Ministry of Education Engineering Research Center of Bioreactor and Pharmaceutical Development, Jilin Agricultural University, Changchun, China aut NATIONALLICENCE 700 1- Jiang Chao Ministry of Education Engineering Research Center of Bioreactor and Pharmaceutical Development, Jilin Agricultural University, Changchun, China aut NATIONALLICENCE 700 1- Li Xiaokun College of Life Sciences, Jilin University, 130021, Changchun, China aut NATIONALLICENCE 773 0- Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/16(2015-08-01), 6753-6764 0175-7598 99:16<6753 2015 99 253