title: Tobacco seeds as efficient production platform for a biologically active anti-HBsAg monoclonal antibody.
abstract: The use of plants as heterologous hosts is one of the most promising technologies for manufacturing valuable recombinant proteins. Plant seeds, in particular, constitute ideal production platforms for long-term applications requiring a steady supply of starting material, as they combine the general advantages of plants as bioreactors with the possibility of biomass storage for long periods in a relatively small volume, thus allowing manufacturers to decouple upstream and downstream processing. In the present work we have used transgenic tobacco seeds to produce large amounts of a functionally active mouse monoclonal antibody against the Hepatitis B Virus surface antigen, fused to a KDEL endoplasmic reticulum retrieval motif, under control of regulatory sequences from common bean (Phaseolus vulgaris) seed storage proteins. The antibody accumulated to levels of 6.5 mg/g of seed in the T3 generation, and was purified by Protein A affinity chromatography combined with SEC-HPLC. N-glycan analysis indicated that, despite the KDEL signal, the seed-derived plantibody bore both high-mannose and complex-type sugars that indicate partial passage through the Golgi compartment, although its performance in the immunoaffinity purification of HBsAg was unaffected. An analysis discussing the industrial feasibility of replacing the currently used tobacco leaf-derived plantibody with this seed-derived variant is also presented.
authors.affiliation: Plant Biotechnology Department, Center for Genetic Engineering and Biotechnology (CIGB), PO Box 6162, 10600, Havana, Havana, Cuba. abel.hernandez@cigb.edu.cu.
authors.forename: Abel
authors.initials: A
authors.lastname: Hernández-Velázquez
authors.affiliation: Plant Biotechnology Department, Center for Genetic Engineering and Biotechnology (CIGB), PO Box 6162, 10600, Havana, Havana, Cuba.
authors.forename: Alina
authors.initials: A
authors.lastname: López-Quesada
authors.affiliation: Plant Biotechnology Department, Center for Genetic Engineering and Biotechnology (CIGB), PO Box 6162, 10600, Havana, Havana, Cuba.
authors.forename: Yanaysi
authors.initials: Y
authors.lastname: Ceballo-Cámara
authors.affiliation: Department of Carbohydrate Chemistry, Center for Genetic Engineering and Biotechnology (CIGB), Havana, Cuba.
authors.forename: Gleysin
authors.initials: G
authors.lastname: Cabrera-Herrera
authors.affiliation: Plant Biotechnology Department, Center for Genetic Engineering and Biotechnology (CIGB), PO Box 6162, 10600, Havana, Havana, Cuba.
authors.forename: Kenia
authors.initials: K
authors.lastname: Tiel-González
authors.affiliation: Plant Biotechnology Department, Center for Genetic Engineering and Biotechnology (CIGB), PO Box 6162, 10600, Havana, Havana, Cuba.
authors.forename: Liliana
authors.initials: L
authors.lastname: Mirabal-Ortega
authors.affiliation: Plant Biotechnology Department, Center for Genetic Engineering and Biotechnology (CIGB), PO Box 6162, 10600, Havana, Havana, Cuba.
authors.forename: Marlene
authors.initials: M
authors.lastname: Pérez-Martínez
authors.affiliation: Plant Biotechnology Department, Center for Genetic Engineering and Biotechnology (CIGB), PO Box 6162, 10600, Havana, Havana, Cuba.
authors.forename: Rosabel
authors.initials: R
authors.lastname: Pérez-Castillo
authors.affiliation: Plant Biotechnology Department, Center for Genetic Engineering and Biotechnology (CIGB), PO Box 6162, 10600, Havana, Havana, Cuba.
authors.forename: Yamilka
authors.initials: Y
authors.lastname: Rosabal-Ayán
authors.affiliation: Plant Biotechnology Department, Center for Genetic Engineering and Biotechnology (CIGB), PO Box 6162, 10600, Havana, Havana, Cuba.
authors.forename: Osmani
authors.initials: O
authors.lastname: Ramos-González
authors.affiliation: Plant Biotechnology Department, Center for Genetic Engineering and Biotechnology (CIGB), PO Box 6162, 10600, Havana, Havana, Cuba.
authors.forename: Gil
authors.initials: G
authors.lastname: Enríquez-Obregón
authors.affiliation: Department of Plant Systems Biology, VIB, Plant-made Antibodies and Immunogens, Ghent, Belgium.
authors.forename: Ann
authors.initials: A
authors.lastname: Depicker
authors.affiliation: Plant Biotechnology Department, Center for Genetic Engineering and Biotechnology (CIGB), PO Box 6162, 10600, Havana, Havana, Cuba.
authors.forename: Merardo
authors.initials: M
authors.lastname: Pujol-Ferrer
mesh_terms: D002851:Chromatography, High Pressure Liquid; D002853:Chromatography, Liquid / Q000379:methods; D004591:Electrophoresis, Polyacrylamide Gel; D006514:Hepatitis B Surface Antigens / Q000276:immunology* / Q000302:isolation & purification; D046549:Plantibodies / Q000276:immunology*; D012639:Seeds / Q000276:immunology*; D014026:Nicotiana / Q000196:embryology*
chemical_list: D006514:Hepatitis B Surface Antigens; D046549:Plantibodies
keywords: Hepatitis B; Monoclonal antibody; N-glycan; Phaseolin promoter; Protein expression; Tobacco seeds