期刊
PROCESS BIOCHEMISTRY
卷 45, 期 11, 页码 1816-1820出版社
ELSEVIER SCI LTD
DOI: 10.1016/j.procbio.2010.01.016
关键词
Zera; Recombinant protein; Protein body-like; Eukaryote cells; Protein production; Downstream processing
资金
- Ministerio de Ciencia e Innovacion
- AGAUR [2006FI02015]
- CIDEM
- Generalitat de Catalunya [CeRBA-SGR00182]
- ERA Biotech
In this study, we describe a process for protein expression and purification from plants and insect cells based on the accumulation of recombinant proteins in protein bodies. This technology is using Zera (R), which sequence has the capacity to trigger in vivo the formation of dense, non-secretory storage protein body-like organelles derived from the endoplasmic reticulum (ER). With this method, recombinant human growth hormone (hGH) was expressed and purified from protein bodies accumulated in plants (Nicotiana benthamiana) and in insect cells (Spodoptera frugiperda). We found that recombinant Zera-hGH are stored in large quantity inside those proteins bodies and can be easily recovered during a one-step process from plant and insect cell biomass. After solubilization of recombinant protein bodies and cleavage of Zera tag from the fusion protein, active hGH was finally purified by a single chromatography step. These results indicate that recombinant proteins derived from Zera-fusion could provide both an efficient protein production system and eased purification downstream process. (C) 2010 Elsevier Ltd. All rights reserved.
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