title: Application of electrochemically produced aluminum hydroxide gel for prepurification of recombinant synthetic green fluorescent protein produced in tobacco leaves.
abstract: The use of recombinant proteins has increased greatly in recent years, as also have increased the number of techniques and materials used for their production and purification. Among the different types of bioreactors being studied, there is a general consensus among scientists that production in green plant tissues such as leaves is more feasible. However, the presence of chlorophyll and phenolic compounds in plant extracts, which can precipitate and denature the proteins besides damaging separation membranes and gels, makes this technology impracticable on a commercial scale. In the present work, the adsorption to electrochemically produced aluminum hydroxide gel was applied as a prepurification step for recombinant synthetic green fluorescent protein (sGFP), also referred to as enhanced green fluorescent protein, produced in Nicotiana benthamiana leaves. Removal efficiencies of 99.7% of chlorophyll, 88.5% of phenolic compounds, and 38.5% of native proteins from the N. benthamiana extracts were achieved without removing sGFP from the extracts. As electrochemical preparation of aluminum hydroxide gel is a cost-effective technique, its use can substantially contribute to the development of future production platforms for recombinant proteins produced in green plant tissues of pharmaceutical and industrial interest.
authors.affiliation: Departamento de Processos Biotecnológicos, Faculdade de Engenharia Química, Universidade Estadual de Campinas, CP 6066, CEP 13083-970, Campinas, SP, Brazil. goran.robic@sandoz.com
authors.forename: Goran
authors.initials: G
authors.lastname: Robić
authors.forename: Cristiano
authors.initials: C
authors.lastname: Lacorte
authors.forename: Elibio L
authors.initials: EL
authors.lastname: Rech
authors.forename: Everson A
authors.initials: EA
authors.lastname: Miranda
mesh_terms: D000536:Aluminum Hydroxide / Q000737:chemistry*; D002734:Chlorophyll / Q000737:chemistry; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism*; D007525:Isoelectric Focusing; D010636:Phenols / Q000737:chemistry; D010936:Plant Extracts / Q000737:chemistry; D018515:Plant Leaves / Q000235:genetics / Q000378:metabolism*; D030821:Plants, Genetically Modified / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism*; D014026:Nicotiana / Q000235:genetics / Q000378:metabolism*
chemical_list: D010636:Phenols; D010936:Plant Extracts; D011994:Recombinant Proteins; C485184:enhanced green fluorescent protein; D002734:Chlorophyll; D049452:Green Fluorescent Proteins; D000536:Aluminum Hydroxide