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

    Author(s)
    Sainsbury, Frank
    Jutras, Philippe V
    Vorster, Juan
    Goulet, Marie-Claire
    Michaud, Dominique
    Griffith University Author(s)
    Sainsbury, Frank
    Year published
    2016
    Metadata
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    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 ...
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    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 hostNicotiana 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 inN. 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.
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    Journal Title
    Frontiers in Plant Science
    Volume
    7
    DOI
    https://doi.org/10.3389/fpls.2016.00141
    Subject
    Plant biology
    Science & Technology
    Life Sciences & Biomedicine
    Plant Sciences
    plant molecular farming
    protein purification
    Publication URI
    http://hdl.handle.net/10072/409475
    Collection
    • Journal articles

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