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

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 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

Fingerprint

cell culture
extracts
proteins
water
continuous systems
recombinant proteins
plant extracts
cell suspension culture
concentrates
hydrophobins
industry
methodology

Keywords

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

Cite this

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
Reuter, Lauri ; Conley, Andrew ; Joensuu, Jussi. / Continuous flow separation of hydrophobin fusion proteins from plant cell culture extract. Recombinant Proteins from Plants: Methods and Protocols. editor / Jacqueline MacDonald ; Igor Kolotilin ; Rima Menassa. Second. ed. Springer, 2016. pp. 189-197 (Methods in Molecular Biology, Vol. 1385).
@inbook{a791c4ecf98c4bb995ef7e974edaacb9,
title = "Continuous flow separation of hydrophobin fusion proteins from plant cell culture extract",
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.",
keywords = "continuous aqueous two-phase system, ATPS, hydrophobin, HFBI, protein purification, protein fusion",
author = "Lauri Reuter and Andrew Conley and Jussi Joensuu",
year = "2016",
doi = "10.1007/978-1-4939-3289-4_14",
language = "English",
isbn = "978-1-4939-3288-7",
series = "Methods in Molecular Biology",
publisher = "Springer",
pages = "189--197",
editor = "Jacqueline MacDonald and Igor Kolotilin and Rima Menassa",
booktitle = "Recombinant Proteins from Plants",
address = "Germany",
edition = "Second",

}

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 edn, Springer, Methods in Molecular Biology, vol. 1385, pp. 189-197. https://doi.org/10.1007/978-1-4939-3289-4_14

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 proceedingChapter or book articleProfessional

TY - CHAP

T1 - Continuous flow separation of hydrophobin fusion proteins from plant cell culture extract

AU - Reuter, Lauri

AU - Conley, Andrew

AU - Joensuu, Jussi

PY - 2016

Y1 - 2016

N2 - 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.

AB - 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.

KW - continuous aqueous two-phase system

KW - ATPS

KW - hydrophobin

KW - HFBI

KW - protein purification

KW - protein fusion

U2 - 10.1007/978-1-4939-3289-4_14

DO - 10.1007/978-1-4939-3289-4_14

M3 - Chapter or book article

SN - 978-1-4939-3288-7

T3 - Methods in Molecular Biology

SP - 189

EP - 197

BT - Recombinant Proteins from Plants

A2 - MacDonald, Jacqueline

A2 - Kolotilin, Igor

A2 - Menassa, Rima

PB - Springer

ER -

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