Expression of bioactive anti-CD20 antibody fragments and induction of ER stress response in Arabidopsis seeds

Verfasser / Beitragende:
[Dezhong Wang, Jisheng Ma, Difei Sun, Haiyan Li, Chao Jiang, Xiaokun Li]
Ort, Verlag, Jahr:
2015
Enthalten in:
Applied Microbiology and Biotechnology, 99/16(2015-08-01), 6753-6764
Format:
Artikel (online)
ID: 605498482
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024 7 0 |a 10.1007/s00253-015-6601-6  |2 doi 
035 |a (NATIONALLICENCE)springer-10.1007/s00253-015-6601-6 
245 0 0 |a Expression of bioactive anti-CD20 antibody fragments and induction of ER stress response in Arabidopsis seeds  |h [Elektronische Daten]  |c [Dezhong Wang, Jisheng Ma, Difei Sun, Haiyan Li, Chao Jiang, Xiaokun Li] 
520 3 |a 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. 
540 |a Springer-Verlag Berlin Heidelberg, 2015 
690 7 |a Arabidopsis seeds  |2 nationallicence 
690 7 |a Anti-CD20 antibody fragments  |2 nationallicence 
690 7 |a ADCC  |2 nationallicence 
690 7 |a CDC  |2 nationallicence 
690 7 |a ER stress response  |2 nationallicence 
690 7 |a ER-associated degradation  |2 nationallicence 
700 1 |a Wang  |D Dezhong  |u College of Life Sciences, Jilin University, 130021, Changchun, China  |4 aut 
700 1 |a Ma  |D Jisheng  |u School of Pharmaceutical Sciences, Zhejiang Key Laboratory of Biotechnology and Pharmaceutical Engineering, Wenzhou Medical University, Chashan College Park, 325035, Wenzhou, China  |4 aut 
700 1 |a Sun  |D Difei  |u School of Pharmaceutical Sciences, Zhejiang Key Laboratory of Biotechnology and Pharmaceutical Engineering, Wenzhou Medical University, Chashan College Park, 325035, Wenzhou, China  |4 aut 
700 1 |a Li  |D Haiyan  |u Ministry of Education Engineering Research Center of Bioreactor and Pharmaceutical Development, Jilin Agricultural University, Changchun, China  |4 aut 
700 1 |a Jiang  |D Chao  |u Ministry of Education Engineering Research Center of Bioreactor and Pharmaceutical Development, Jilin Agricultural University, Changchun, China  |4 aut 
700 1 |a Li  |D Xiaokun  |u College of Life Sciences, Jilin University, 130021, Changchun, China  |4 aut 
773 0 |t Applied Microbiology and Biotechnology  |d Springer Berlin Heidelberg  |g 99/16(2015-08-01), 6753-6764  |x 0175-7598  |q 99:16<6753  |1 2015  |2 99  |o 253 
856 4 0 |u https://doi.org/10.1007/s00253-015-6601-6  |q text/html  |z Onlinezugriff via DOI 
898 |a BK010053  |b XK010053  |c XK010000 
900 7 |a Metadata rights reserved  |b Springer special CC-BY-NC licence  |2 nationallicence 
908 |D 1  |a research-article  |2 jats 
949 |B NATIONALLICENCE  |F NATIONALLICENCE  |b NL-springer 
950 |B NATIONALLICENCE  |P 856  |E 40  |u https://doi.org/10.1007/s00253-015-6601-6  |q text/html  |z Onlinezugriff via DOI 
950 |B NATIONALLICENCE  |P 700  |E 1-  |a Wang  |D Dezhong  |u College of Life Sciences, Jilin University, 130021, Changchun, China  |4 aut 
950 |B NATIONALLICENCE  |P 700  |E 1-  |a Ma  |D Jisheng  |u School of Pharmaceutical Sciences, Zhejiang Key Laboratory of Biotechnology and Pharmaceutical Engineering, Wenzhou Medical University, Chashan College Park, 325035, Wenzhou, China  |4 aut 
950 |B NATIONALLICENCE  |P 700  |E 1-  |a Sun  |D Difei  |u School of Pharmaceutical Sciences, Zhejiang Key Laboratory of Biotechnology and Pharmaceutical Engineering, Wenzhou Medical University, Chashan College Park, 325035, Wenzhou, China  |4 aut 
950 |B NATIONALLICENCE  |P 700  |E 1-  |a Li  |D Haiyan  |u Ministry of Education Engineering Research Center of Bioreactor and Pharmaceutical Development, Jilin Agricultural University, Changchun, China  |4 aut 
950 |B NATIONALLICENCE  |P 700  |E 1-  |a Jiang  |D Chao  |u Ministry of Education Engineering Research Center of Bioreactor and Pharmaceutical Development, Jilin Agricultural University, Changchun, China  |4 aut 
950 |B NATIONALLICENCE  |P 700  |E 1-  |a Li  |D Xiaokun  |u College of Life Sciences, Jilin University, 130021, Changchun, China  |4 aut 
950 |B NATIONALLICENCE  |P 773  |E 0-  |t Applied Microbiology and Biotechnology  |d Springer Berlin Heidelberg  |g 99/16(2015-08-01), 6753-6764  |x 0175-7598  |q 99:16<6753  |1 2015  |2 99  |o 253