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.
authors.affiliation: VTT Technical Research Centre of Finland, Industrial Biotechnology, Tietotie 2, Espoo, 1000, 02044 VTT, Finland.
authors.forename: Lauri J
authors.initials: LJ
authors.lastname: Reuter
authors.affiliation: VTT Technical Research Centre of Finland, Industrial Biotechnology, Tietotie 2, Espoo, 1000, 02044 VTT, Finland.
authors.forename: Andrew J
authors.initials: AJ
authors.lastname: Conley
authors.affiliation: VTT Technical Research Centre of Finland, Industrial Biotechnology, Tietotie 2, Espoo, 1000, 02044 VTT, Finland. jussi.joensuu@vtt.fi.
authors.forename: Jussi J
authors.initials: JJ
authors.lastname: Joensuu
mesh_terms: D018929:Cell Culture Techniques; D005656:Fungal Proteins / Q000302:isolation & purification*; D049452:Green Fluorescent Proteins / Q000235:genetics; D059625:Liquid-Liquid Extraction; D059828:Plant Cells; D030821:Plants, Genetically Modified; D011993:Recombinant Fusion Proteins / Q000302:isolation & purification*; D014026:Nicotiana / Q000235:genetics* / Q000378:metabolism
chemical_list: D005656:Fungal Proteins; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins
keywords: ATPS; Continuous aqueous two-phase system; HFBI; Hydrophobin; Protein fusion; Protein purification