A chimeric affinity tag for efficient expression and chromatographic purification of heterologous proteins from plants

Sainsbury, Frank, Jutras, Philippe V., Vorster, Juan, Goulet, Marie-Claire and Michaud, Dominique (2016) A chimeric affinity tag for efficient expression and chromatographic purification of heterologous proteins from plants. Frontiers in Plant Science, 7 FEB2016: . doi:10.3389/fpls.2016.00141


Author Sainsbury, Frank
Jutras, Philippe V.
Vorster, Juan
Goulet, Marie-Claire
Michaud, Dominique
Title A chimeric affinity tag for efficient expression and chromatographic purification of heterologous proteins from plants
Journal name Frontiers in Plant Science   Check publisher's open access policy
ISSN 1664-462X
Publication date 2016-02-15
Year available 2016
Sub-type Article (original research)
DOI 10.3389/fpls.2016.00141
Open Access Status DOI
Volume 7
Issue FEB2016
Total pages 11
Place of publication Lausanne, Switzerland
Publisher Frontiers Research Foundation
Collection year 2017
Language eng
Formatted abstract
The use of plants as expression hosts for recombinant proteins is an increasingly attractive option for the production of complex and challenging biopharmaceuticals. Tools are needed at present to marry recent developments in high-yielding gene vectors for heterologous expression with routine protein purification techniques. In this study, we designed the Cysta-tag, a new purification tag for immobilized metal affinity chromatography (IMAC) of plant-made proteins based on the protein-stabilizing fusion partner SlCYS8. We show that the Cysta-tag may be used to readily purify proteins under native conditions, and then be removed enzymatically to isolate the protein of interest. We also show that commonly used protease recognition sites for linking purification tags are differentially stable in leaves of the commonly used expression host Nicotiana benthamiana, with those linkers susceptible to cysteine proteases being less stable then serine protease-cleavable linkers. As an example, we describe a Cysta-tag experimental scheme for the one-step purification of a clinically useful protein, human α1-antitrypsin, transiently expressed in N. benthamiana. With potential applicability to the variety of chromatography formats commercially available for IMAC-based protein purification, the Cysta-tag provides a convenient means for the efficient and cost-effective purification of recombinant proteins from plant tissues.
Keyword Human α1-antitrypsin
Immobilized metal affinity chromatography
Plant molecular farming
Protein purification
Tomato cystatin SlCYS8
Q-Index Code C1
Q-Index Status Provisional Code
Institutional Status UQ

Document type: Journal Article
Sub-type: Article (original research)
Collections: HERDC Pre-Audit
Australian Institute for Bioengineering and Nanotechnology Publications
 
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