Continuous flow separation of hydrophobin fusion proteins from plant cell culture extract

Lauri Reuter, Andrew Conley, Jussi Joensuu (Corresponding author)

    Research output: Chapter in Book/Report/Conference proceedingChapter or book articleProfessional

    2 Citations (Scopus)

    Abstract

    Fusion to fungal hydrophobins has proven to be a useful tool to enhance accumulation and recovery of recombinant proteins in plants. Aqueous two-phase separation (ATPS) is an attractive system to capture hydrophobin fusion proteins from plant extracts. The process can simultaneously purify and concentrate target protein with minimal background. ATPS avoids the use of chromatographic column steps, can be carried out in a short time frame, and is amenable to industrial-scale protein purification. A drawback of performing ATPS in large volumes is the lengthy time required for phase separation; however, this can be avoided by incorporating continuous systems, which are often preferred by the processing industry. This method chapter illustrates the capture of GFP-HFBI hydrophobin fusion protein from BY-2 plant cell suspension extract using a semi-continuous ATPS method.
    Original languageEnglish
    Title of host publicationRecombinant Proteins from Plants
    Subtitle of host publicationMethods and Protocols
    EditorsJacqueline MacDonald, Igor Kolotilin, Rima Menassa
    PublisherSpringer
    Pages189-197
    EditionSecond
    ISBN (Electronic)978-1-4939-3289-4
    ISBN (Print)978-1-4939-3288-7
    DOIs
    Publication statusPublished - 2016
    MoE publication typeD2 Article in professional manuals or guides or professional information systems or text book material

    Publication series

    SeriesMethods in Molecular Biology
    Volume1385
    ISSN1064-3745

    Keywords

    • continuous aqueous two-phase system
    • ATPS
    • hydrophobin
    • HFBI
    • protein purification
    • protein fusion

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