Bioseparation of Recombinant Proteins from Plant Extract with Hydrophobin Fusion Technology

Jussi Joensuu (Corresponding author), Andrew Conley, Markus Linder, Rima Menassa

    Research output: Chapter in Book/Report/Conference proceedingChapter or book articleScientificpeer-review

    16 Citations (Scopus)

    Abstract

    Two main hurdles hinder the widespread acceptance of plants as a preferred protein expression platform: low accumulation levels and expensive chromatographic purification methods. Fusion of proteins of interest to fungal hydrophobins has provided a tool to address both accumulation and purification issues. In this method, we describe the one-step purification of a GFP-HFBI fusion from crude plant extract using an aqueous two-phase system (ATPS). ATPS can be carried out in a very short time frame, yields relatively pure protein with very few contaminants, and does not require any chromatographic column steps. This purification system takes advantage of the affinity of hydrophobins to the micellar phase of widely available nonionic surfactants, such as Triton X-114, and can be easily scaled up for industrial-scale protein purification.
    Original languageEnglish
    Title of host publicationRecombinant Gene Expression
    Subtitle of host publicationReviews and Protocols
    EditorsArgelia Lorence
    Place of PublicationTotowa
    PublisherHumana Press
    Chapter28
    Pages527-534
    ISBN (Electronic)978-1-61779-433-9
    ISBN (Print)978-1-61779-432-2, 978-1-4939-6221-1
    DOIs
    Publication statusPublished - 2012
    MoE publication typeA3 Part of a book or another research book

    Publication series

    SeriesMethods in Molecular Biology
    Volume824
    ISSN1064-3745

    Keywords

    • Aqueous two-phase system
    • ATPS
    • Hydrophobin
    • HFBI
    • Protein purification
    • Protein fusion
    • Molecular farming

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