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

Lauri Reuter; Andrew Conley; Jussi Joensuu

doi:10.1007/978-1-4939-3289-4_14

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

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

BA3402 Protein production

Research output: Chapter in Book/Report/Conference proceeding › Chapter or book article › Professional

1 Citation (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 language	English
Title of host publication	Recombinant Proteins from Plants
Subtitle of host publication	Methods and Protocols
Editors	Jacqueline MacDonald, Igor Kolotilin, Rima Menassa
Publisher	Springer
Pages	189-197
Edition	Second
ISBN (Electronic)	978-1-4939-3289-4
ISBN (Print)	978-1-4939-3288-7
DOIs	https://doi.org/10.1007/978-1-4939-3289-4_14
Publication status	Published - 2016
MoE publication type	D2 Article in professional manuals or guides or professional information systems or text book material

Publication series

Series	Methods in Molecular Biology
Volume	1385
ISSN	1064-3745

Keywords

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

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Reuter, L., Conley, A., & Joensuu, J. (2016). Continuous flow separation of hydrophobin fusion proteins from plant cell culture extract. In J. MacDonald, I. Kolotilin, & R. Menassa (Eds.), Recombinant Proteins from Plants: Methods and Protocols (Second ed., pp. 189-197). Springer. Methods in Molecular Biology, Vol.. 1385 https://doi.org/10.1007/978-1-4939-3289-4_14

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Continuous flow separation of hydrophobin fusion proteins from plant cell culture extract. / Reuter, Lauri; Conley, Andrew; Joensuu, Jussi (Corresponding author).

Recombinant Proteins from Plants: Methods and Protocols. ed. / Jacqueline MacDonald; Igor Kolotilin; Rima Menassa. Second. ed. Springer, 2016. p. 189-197 (Methods in Molecular Biology, Vol. 1385).

Research output: Chapter in Book/Report/Conference proceeding › Chapter or book article › Professional

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