title: Comparative in vitro and experimental in vivo studies of the anti-epidermal growth factor receptor antibody nimotuzumab and its aglycosylated form produced in transgenic tobacco plants.
abstract: A broad variety of foreign genes can be expressed in transgenic plants, which offer the opportunity for large-scale production of pharmaceutical proteins, such as therapeutic antibodies. Nimotuzumab is a humanized anti-epidermal growth factor receptor (EGFR) recombinant IgG1 antibody approved in different countries for the treatment of head and neck squamous cell carcinoma, paediatric and adult glioma, and nasopharyngeal and oesophageal cancers. Because the antitumour mechanism of nimotuzumab is mainly attributed to its ability to interrupt the signal transduction cascade triggered by EGF/EGFR interaction, we have hypothesized that an aglycosylated form of this antibody, produced by mutating the N(297)  position in the IgG(1)  Fc region gene, would have similar biochemical and biological properties as the mammalian-cell-produced glycosylated counterpart. In this paper, we report the production and characterization of an aglycosylated form of nimotuzumab in transgenic tobacco plants. The comparison of the plantibody and nimotuzumab in terms of recognition of human EGFR, effect on tyrosine phosphorylation and proliferation in cells in response to EGF, competition with radiolabelled EGF for EGFR, affinity measurements of Fab fragments, pharmacokinetic and biodistribution behaviours in rats and antitumour effects in nude mice bearing human A431 tumours showed that both antibody forms have very similar in vitro and in vivo properties. Our results support the idea that the production of aglycosylated forms of some therapeutic antibodies in transgenic plants is a feasible approach when facing scaling strategies for anticancer immunoglobulins.
authors.affiliation: Center for Genetic Engineering and Biotechnology (CIGB), Playa, Havana, Cuba. meilyn.rodriguez@cigb.edu.cu
authors.forename: Meilyn
authors.initials: M
authors.lastname: Rodríguez
authors.forename: Lincidio
authors.initials: L
authors.lastname: Pérez
authors.forename: Jorge V
authors.initials: JV
authors.lastname: Gavilondo
authors.forename: Greta
authors.initials: G
authors.lastname: Garrido
authors.forename: Mónica
authors.initials: M
authors.lastname: Bequet-Romero
authors.forename: Ignacio
authors.initials: I
authors.lastname: Hernández
authors.forename: Vivian
authors.initials: V
authors.lastname: Huerta
authors.forename: Gleysin
authors.initials: G
authors.lastname: Cabrera
authors.forename: Marlene
authors.initials: M
authors.lastname: Pérez
authors.forename: Osmani
authors.initials: O
authors.lastname: Ramos
authors.forename: René
authors.initials: R
authors.lastname: Leyva
authors.forename: Mariela
authors.initials: M
authors.lastname: León
authors.forename: Pedro Luis
authors.initials: PL
authors.lastname: Ramos
authors.forename: Ada
authors.initials: A
authors.lastname: Triguero
authors.forename: Abel
authors.initials: A
authors.lastname: Hernández
authors.forename: Belinda
authors.initials: B
authors.lastname: Sánchez
authors.forename: Marta
authors.initials: M
authors.lastname: Ayala
authors.forename: Jeny
authors.initials: J
authors.lastname: Soto
authors.forename: Ernesto
authors.initials: E
authors.lastname: González
authors.forename: Osmani
authors.initials: O
authors.lastname: Mendoza
authors.forename: Kenia
authors.initials: K
authors.lastname: Tiel
authors.forename: Merardo
authors.initials: M
authors.lastname: Pujol
mesh_terms: D000818:Animals; D061067:Antibodies, Monoclonal, Humanized / Q000096:biosynthesis* / Q000494:pharmacology*; D000970:Antineoplastic Agents / Q000378:metabolism; D045744:Cell Line, Tumor; D066246:ErbB Receptors / Q000037:antagonists & inhibitors*; D005260:Female; D006801:Humans; D007136:Immunoglobulins / Q000096:biosynthesis*; D051379:Mice; D008819:Mice, Nude; D058634:Molecular Farming / Q000379:methods; D010766:Phosphorylation / Q000187:drug effects; D046549:Plantibodies / Q000378:metabolism / Q000494:pharmacology*; D030821:Plants, Genetically Modified / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar; D011994:Recombinant Proteins; D014026:Nicotiana / Q000235:genetics* / Q000378:metabolism*; D014443:Tyrosine / Q000378:metabolism
chemical_list: D061067:Antibodies, Monoclonal, Humanized; D000970:Antineoplastic Agents; D007136:Immunoglobulins; D046549:Plantibodies; D011994:Recombinant Proteins; D014443:Tyrosine; C501466:nimotuzumab; D066246:ErbB Receptors