title: Expression and characterization of an anti-(hepatitis B surface antigen) glycosylated mouse antibody in transgenic tobacco (Nicotiana tabacum) plants and its use in the immunopurification of its target antigen.
abstract: Transgenic plants expressing recombinant immunoglobulins have arisen as an alternative technology for the large-scale production of antibodies useful in therapeutics and in industrial processes. In the present paper we report the expression in transgenic tobacco ( Nicotiana tabacum ) of an anti-HBsAg [anti-(hepatitis B virus surface antigen)] mouse IgG1 mAb (monoclonal antibody), currently used for the industrial purification of the recombinant vaccine antigen. Using the sweet potato sporamin signal peptide, a KDEL (Lys-Asp-Glu-Leu) ER (endoplasmic reticulum) anchorage domain, and a heavy- and light-chain gene tandem construction, we generated F1 plants in which the expression of the antibody accounted for 0.5% of the total soluble proteins. The 'plantibody' (functional IgG antibody produced in plants) was easily purified by Protein A-Sepharose chromatography with a yield of approximately 35 microg/g of fresh leaf material, and its glycosylation indicated that, irrespective of the KDEL signal, the molecule is modified in both the ER and Golgi. Finally, a successful comparison of the plantibody with the ascites-derived mAb in the immunoaffinity purification of the vaccine recombinant HBsAg was performed. Taken as a whole, our results show that the large-scale production of this antibody of industrial relevance in transgenic tobacco is feasible.
authors.affiliation: Center for Genetic Engineering and Biotechnology, P.O. Box 6162, Havana 10600, Cuba.
authors.forename: Nadia
authors.initials: N
authors.lastname: Ramírez
authors.forename: Meilyn
authors.initials: M
authors.lastname: Rodríguez
authors.forename: Marta
authors.initials: M
authors.lastname: Ayala
authors.forename: José
authors.initials: J
authors.lastname: Cremata
authors.forename: Marlene
authors.initials: M
authors.lastname: Pérez
authors.forename: Anazuria
authors.initials: A
authors.lastname: Martínez
authors.forename: Marbelis
authors.initials: M
authors.lastname: Linares
authors.forename: Yasser
authors.initials: Y
authors.lastname: Hevia
authors.forename: Rolando
authors.initials: R
authors.lastname: Páez
authors.forename: Rodolfo
authors.initials: R
authors.lastname: Valdés
authors.forename: Jorge V
authors.initials: JV
authors.lastname: Gavilondo
authors.forename: Guillermo
authors.initials: G
authors.lastname: Selman-Housein
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D003001:Cloning, Molecular / Q000379:methods; D018506:Gene Expression Regulation, Plant / Q000502:physiology; D006031:Glycosylation; D006510:Hepatitis B Antibodies / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification; D007118:Immunoassay / Q000379:methods; D051379:Mice / Q000235:genetics; D008807:Mice, Inbred BALB C; D018515:Plant Leaves / Q000235:genetics / Q000378:metabolism; D030821:Plants, Genetically Modified / Q000378:metabolism*; D015202:Protein Engineering / Q000379:methods; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000302:isolation & purification; D014026:Nicotiana / Q000235:genetics* / Q000378:metabolism*; D014614:Vaccines, Synthetic / Q000737:chemistry
chemical_list: D000911:Antibodies, Monoclonal; D006510:Hepatitis B Antibodies; D011994:Recombinant Proteins; D014614:Vaccines, Synthetic