title: Challenges in electrochemical pre-purification of recombinant proteins from green plant tissues: sgfp produced in tobacco leaves.
abstract: The use of recombinant proteins has increased greatly in recent years, as have the number of techniques and materials used for their production and purification. The principal advantage of using plants as bioreactors is the cost of the recombinant protein production, which is about 1000-fold lower as in the case of using CHO cells commonly applied in industry today. Among the different types of "green" bioreactors being studied today, 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. Electrochemically produced aluminium hydroxide gel can be used to adsorb these compounds, and pre-purify recombinant synthetic green fluorescent protein (sGFP) 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. Since 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, Campinas, SP, Brazil. goran.robic@sandoz.com
authors.forename: Goran
authors.initials: G
authors.lastname: Robić
mesh_terms: D002734:Chlorophyll / Q000378:metabolism; D055664:Electrochemical Techniques / Q000379:methods*; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D010636:Phenols / Q000378:metabolism; D018515:Plant Leaves / Q000235:genetics / Q000378:metabolism*; D030821:Plants, Genetically Modified / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism*; D014026:Nicotiana / Q000235:genetics / Q000378:metabolism*
chemical_list: D010636:Phenols; D011994:Recombinant Proteins; D002734:Chlorophyll; D049452:Green Fluorescent Proteins