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

Abstract 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 ret...
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Autor*in:	Wang, Dezhong [verfasserIn] Ma, Jisheng Sun, Difei Li, Haiyan Jiang, Chao Li, Xiaokun

Format:	Artikel
Sprache:	Englisch

Erschienen:	2015

Schlagwörter:	seeds Anti-CD20 antibody fragments ADCC CDC ER stress response ER-associated degradation

Anmerkung:	© Springer-Verlag Berlin Heidelberg 2015

Übergeordnetes Werk:	Enthalten in: Applied microbiology and biotechnology - Springer Berlin Heidelberg, 1984, 99(2015), 16 vom: 09. Mai, Seite 6753-6764
Übergeordnetes Werk:	volume:99 ; year:2015 ; number:16 ; day:09 ; month:05 ; pages:6753-6764

Links:	Volltext

DOI / URN:	10.1007/s00253-015-6601-6

Katalog-ID:	OLC2050769709

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allfields	10.1007/s00253-015-6601-6 doi (DE-627)OLC2050769709 (DE-He213)s00253-015-6601-6-p DE-627 ger DE-627 rakwb eng 570 VZ 12 ssgn BIODIV DE-30 fid Wang, Dezhong verfasserin aut Expression of bioactive anti-CD20 antibody fragments and induction of ER stress response in Arabidopsis seeds 2015 Text txt rdacontent ohne Hilfsmittel zu benutzen n rdamedia Band nc rdacarrier © Springer-Verlag Berlin Heidelberg 2015 Abstract 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 13 days 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. seeds Anti-CD20 antibody fragments ADCC CDC ER stress response ER-associated degradation Ma, Jisheng aut Sun, Difei aut Li, Haiyan aut Jiang, Chao aut Li, Xiaokun aut Enthalten in Applied microbiology and biotechnology Springer Berlin Heidelberg, 1984 99(2015), 16 vom: 09. Mai, Seite 6753-6764 (DE-627)129942634 (DE-600)392453-1 (DE-576)015507750 0175-7598 nnns volume:99 year:2015 number:16 day:09 month:05 pages:6753-6764 https://doi.org/10.1007/s00253-015-6601-6 lizenzpflichtig Volltext GBV_USEFLAG_A SYSFLAG_A GBV_OLC FID-BIODIV SSG-OLC-TEC SSG-OLC-CHE SSG-OLC-PHA SSG-OLC-DE-84 GBV_ILN_70 GBV_ILN_130 GBV_ILN_267 GBV_ILN_2018 GBV_ILN_4012 GBV_ILN_4082 GBV_ILN_4277 GBV_ILN_4305 AR 99 2015 16 09 05 6753-6764
spelling	10.1007/s00253-015-6601-6 doi (DE-627)OLC2050769709 (DE-He213)s00253-015-6601-6-p DE-627 ger DE-627 rakwb eng 570 VZ 12 ssgn BIODIV DE-30 fid Wang, Dezhong verfasserin aut Expression of bioactive anti-CD20 antibody fragments and induction of ER stress response in Arabidopsis seeds 2015 Text txt rdacontent ohne Hilfsmittel zu benutzen n rdamedia Band nc rdacarrier © Springer-Verlag Berlin Heidelberg 2015 Abstract 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 13 days 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. seeds Anti-CD20 antibody fragments ADCC CDC ER stress response ER-associated degradation Ma, Jisheng aut Sun, Difei aut Li, Haiyan aut Jiang, Chao aut Li, Xiaokun aut Enthalten in Applied microbiology and biotechnology Springer Berlin Heidelberg, 1984 99(2015), 16 vom: 09. Mai, Seite 6753-6764 (DE-627)129942634 (DE-600)392453-1 (DE-576)015507750 0175-7598 nnns volume:99 year:2015 number:16 day:09 month:05 pages:6753-6764 https://doi.org/10.1007/s00253-015-6601-6 lizenzpflichtig Volltext GBV_USEFLAG_A SYSFLAG_A GBV_OLC FID-BIODIV SSG-OLC-TEC SSG-OLC-CHE SSG-OLC-PHA SSG-OLC-DE-84 GBV_ILN_70 GBV_ILN_130 GBV_ILN_267 GBV_ILN_2018 GBV_ILN_4012 GBV_ILN_4082 GBV_ILN_4277 GBV_ILN_4305 AR 99 2015 16 09 05 6753-6764
allfields_unstemmed	10.1007/s00253-015-6601-6 doi (DE-627)OLC2050769709 (DE-He213)s00253-015-6601-6-p DE-627 ger DE-627 rakwb eng 570 VZ 12 ssgn BIODIV DE-30 fid Wang, Dezhong verfasserin aut Expression of bioactive anti-CD20 antibody fragments and induction of ER stress response in Arabidopsis seeds 2015 Text txt rdacontent ohne Hilfsmittel zu benutzen n rdamedia Band nc rdacarrier © Springer-Verlag Berlin Heidelberg 2015 Abstract 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 13 days 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. seeds Anti-CD20 antibody fragments ADCC CDC ER stress response ER-associated degradation Ma, Jisheng aut Sun, Difei aut Li, Haiyan aut Jiang, Chao aut Li, Xiaokun aut Enthalten in Applied microbiology and biotechnology Springer Berlin Heidelberg, 1984 99(2015), 16 vom: 09. Mai, Seite 6753-6764 (DE-627)129942634 (DE-600)392453-1 (DE-576)015507750 0175-7598 nnns volume:99 year:2015 number:16 day:09 month:05 pages:6753-6764 https://doi.org/10.1007/s00253-015-6601-6 lizenzpflichtig Volltext GBV_USEFLAG_A SYSFLAG_A GBV_OLC FID-BIODIV SSG-OLC-TEC SSG-OLC-CHE SSG-OLC-PHA SSG-OLC-DE-84 GBV_ILN_70 GBV_ILN_130 GBV_ILN_267 GBV_ILN_2018 GBV_ILN_4012 GBV_ILN_4082 GBV_ILN_4277 GBV_ILN_4305 AR 99 2015 16 09 05 6753-6764
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source	Enthalten in Applied microbiology and biotechnology 99(2015), 16 vom: 09. Mai, Seite 6753-6764 volume:99 year:2015 number:16 day:09 month:05 pages:6753-6764
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author	Wang, Dezhong
spellingShingle	Wang, Dezhong ddc 570 ssgn 12 fid BIODIV misc seeds misc Anti-CD20 antibody fragments misc ADCC misc CDC misc ER stress response misc ER-associated degradation Expression of bioactive anti-CD20 antibody fragments and induction of ER stress response in Arabidopsis seeds
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topic_title	570 VZ 12 ssgn BIODIV DE-30 fid Expression of bioactive anti-CD20 antibody fragments and induction of ER stress response in Arabidopsis seeds seeds Anti-CD20 antibody fragments ADCC CDC ER stress response ER-associated degradation
topic	ddc 570 ssgn 12 fid BIODIV misc seeds misc Anti-CD20 antibody fragments misc ADCC misc CDC misc ER stress response misc ER-associated degradation
topic_unstemmed	ddc 570 ssgn 12 fid BIODIV misc seeds misc Anti-CD20 antibody fragments misc ADCC misc CDC misc ER stress response misc ER-associated degradation
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title	Expression of bioactive anti-CD20 antibody fragments and induction of ER stress response in Arabidopsis seeds
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title_full	Expression of bioactive anti-CD20 antibody fragments and induction of ER stress response in Arabidopsis seeds
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author_browse	Wang, Dezhong Ma, Jisheng Sun, Difei Li, Haiyan Jiang, Chao Li, Xiaokun
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title_sort	expression of bioactive anti-cd20 antibody fragments and induction of er stress response in arabidopsis seeds
title_auth	Expression of bioactive anti-CD20 antibody fragments and induction of ER stress response in Arabidopsis seeds
abstract	Abstract 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 13 days 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
abstractGer	Abstract 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 13 days 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
abstract_unstemmed	Abstract 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 13 days 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
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title_short	Expression of bioactive anti-CD20 antibody fragments and induction of ER stress response in Arabidopsis seeds
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