Bioseparation of Recombinant Proteins from Plant Extract with Hydrophobin Fusion Technology

Jussi Joensuu, Andrew Conley, Markus Linder, Rima Menassa

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

13 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
PublisherSpringer
Chapter28
Pages527-534
ISBN (Electronic)978-1-61779-433-9
ISBN (Print)978-1-61779-432-2
DOIs
Publication statusPublished - 2012
MoE publication typeA3 Part of a book or another research book

Fingerprint

recombinant proteins
plant extracts
purification methods
nonionic surfactants
proteins
protein synthesis
water
hydrophobins
methodology

Keywords

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

Cite this

Joensuu, J., Conley, A., Linder, M., & Menassa, R. (2012). Bioseparation of Recombinant Proteins from Plant Extract with Hydrophobin Fusion Technology. In Recombinant Gene Expression: Reviews and Protocols (pp. 527-534). Springer. https://doi.org/10.1007/978-1-61779-433-9_28
Joensuu, Jussi ; Conley, Andrew ; Linder, Markus ; Menassa, Rima. / Bioseparation of Recombinant Proteins from Plant Extract with Hydrophobin Fusion Technology. Recombinant Gene Expression: Reviews and Protocols. Springer, 2012. pp. 527-534
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Joensuu, J, Conley, A, Linder, M & Menassa, R 2012, Bioseparation of Recombinant Proteins from Plant Extract with Hydrophobin Fusion Technology. in Recombinant Gene Expression: Reviews and Protocols. Springer, pp. 527-534. https://doi.org/10.1007/978-1-61779-433-9_28

Bioseparation of Recombinant Proteins from Plant Extract with Hydrophobin Fusion Technology. / Joensuu, Jussi; Conley, Andrew; Linder, Markus; Menassa, Rima.

Recombinant Gene Expression: Reviews and Protocols. Springer, 2012. p. 527-534.

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

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Joensuu J, Conley A, Linder M, Menassa R. Bioseparation of Recombinant Proteins from Plant Extract with Hydrophobin Fusion Technology. In Recombinant Gene Expression: Reviews and Protocols. Springer. 2012. p. 527-534 https://doi.org/10.1007/978-1-61779-433-9_28