title: Improved virus neutralization by plant-produced anti-HIV antibodies with a homogeneous beta1,4-galactosylated N-glycan profile.
abstract: It is well established that proper N-glycosylation significantly influences the efficacy of monoclonal antibodies (mAbs). However, the specific immunological relevance of individual mAb-associated N-glycan structures is currently largely unknown, because of the heterogeneous N-glycan profiles of mAbs when produced in mammalian cells. Here we report on the generation of a plant-based expression platform allowing the efficient production of mAbs with a homogeneous beta1,4-galactosylated N-glycosylation structure, the major N-glycan species present on serum IgG. This was achieved by the expression of a highly active modified version of the human beta1,4-galactosyltransferase in glycoengineered plants lacking plant-specific glycosylation. Moreover, we demonstrate that two anti-human immunodeficiency virus mAbs with fully beta1,4-galactosylated N-glycans display improved virus neutralization potency when compared with other glycoforms produced in plants and Chinese hamster ovary cells. These findings indicate that mAbs containing such homogeneous N-glycan structures should display improved in vivo activities. Our system, using expression of mAbs in tobacco plants engineered for post-translational protein processing, provides a new means of overcoming the two hurdles that limit the therapeutic use of anti-human immunodeficiency virus mAbs in global health initiatives, low biological potency and high production costs.
authors.affiliation: Department of Applied Genetics and Cell Biology, University of Natural Resources and Applied Life Sciences, 1190 Vienna, Austria.
authors.forename: Richard
authors.initials: R
authors.lastname: Strasser
authors.forename: Alexandra
authors.initials: A
authors.lastname: Castilho
authors.forename: Johannes
authors.initials: J
authors.lastname: Stadlmann
authors.forename: Renate
authors.initials: R
authors.lastname: Kunert
authors.forename: Heribert
authors.initials: H
authors.lastname: Quendler
authors.forename: Pia
authors.initials: P
authors.lastname: Gattinger
authors.forename: Jakub
authors.initials: J
authors.lastname: Jez
authors.forename: Thomas
authors.initials: T
authors.lastname: Rademacher
authors.forename: Friedrich
authors.initials: F
authors.lastname: Altmann
authors.forename: Lukas
authors.initials: L
authors.lastname: Mach
authors.forename: Herta
authors.initials: H
authors.lastname: Steinkellner
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000737:chemistry / Q000276:immunology; D016466:CHO Cells; D002460:Cell Line; D006224:Cricetinae; D003412:Cricetulus; D003433:Crosses, Genetic; D005690:Galactose / Q000378:metabolism*; D005822:Genetic Vectors / Q000235:genetics; D006031:Glycosylation; D015483:HIV Antibodies / Q000096:biosynthesis* / Q000737:chemistry / Q000276:immunology*; D015497:HIV-1 / Q000276:immunology*; D006801:Humans; D009154:Mutation / Q000235:genetics; D009500:Neutralization Tests; D046549:Plantibodies / Q000276:immunology*; D011134:Polysaccharides / Q000378:metabolism*; D020033:Protein Isoforms / Q000737:chemistry / Q000378:metabolism; D051381:Rats; D014026:Nicotiana / Q000235:genetics / Q000378:metabolism; D014170:Transformation, Genetic
chemical_list: D000911:Antibodies, Monoclonal; D015483:HIV Antibodies; D046549:Plantibodies; D011134:Polysaccharides; D020033:Protein Isoforms; D005690:Galactose