title: A plant-derived human monoclonal antibody induces an anti-carbohydrate immune response in rabbits.
abstract: A common argument against using plants as a production system for therapeutic proteins is their inability to perform authentic N-glycosylation. A major concern is the presence of beta 1,2-xylose and core alpha 1,3-fucose residues on complex N-glycans as these nonmammalian N-glycan residues may provoke unwanted side effects in humans. In this study we have investigated the potential antigenicity of plant-type N-glycans attached to a human monoclonal antibody (2G12). Using glyco-engineered plant lines as expression hosts, four 2G12 glycoforms differing in the presence/absence of beta 1,2-xylose and core alpha 1,3-fucose were generated. Systemic immunization of rabbits with a xylose and fucose carrying 2G12 glycoform resulted in a humoral immune response to both N-glycan epitopes. Furthermore, IgE immunoblotting with sera derived from allergic patients revealed binding to plant-produced 2G12 carrying core alpha 1,3 fucosylated N-glycan structures. Our results provide evidence for the adverse potential of nonmammalian N-glycan modifications present on monoclonal antibodies produced in plants. This emphasizes the need for the use of glyco-engineered plants lacking any potentially antigenic N-glycan structures for the production of plant-derived recombinant proteins intended for parenteral human application.
authors.affiliation: Department of Chemistry, University of Natural Resources and Applied Life Sciences, Muthgasse 18, 1190 Vienna, Austria.
authors.forename: Chunsheng
authors.initials: C
authors.lastname: Jin
authors.forename: Friedrich
authors.initials: F
authors.lastname: Altmann
authors.forename: Richard
authors.initials: R
authors.lastname: Strasser
authors.forename: Lukas
authors.initials: L
authors.lastname: Mach
authors.forename: Matthias
authors.initials: M
authors.lastname: Schähs
authors.forename: Renate
authors.initials: R
authors.lastname: Kunert
authors.forename: Thomas
authors.initials: T
authors.lastname: Rademacher
authors.forename: Josef
authors.initials: J
authors.lastname: Glössl
authors.forename: Herta
authors.initials: H
authors.lastname: Steinkellner
mesh_terms: D000818:Animals; D000906:Antibodies / Q000097:blood; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000737:chemistry / Q000276:immunology*; D000918:Antibody Specificity / Q000276:immunology; D052179:Antigens, Plant / Q000737:chemistry / Q000276:immunology*; D015153:Blotting, Western; D000080908:Broadly Neutralizing Antibodies; D016466:CHO Cells; D002241:Carbohydrates / Q000737:chemistry / Q000276:immunology*; D002460:Cell Line; D006224:Cricetinae; D003412:Cricetulus; D003429:Cross Reactions; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D005643:Fucose / Q000032:analysis / Q000276:immunology; D015483:HIV Antibodies; D006801:Humans; D007073:Immunoglobulin E / Q000097:blood; D018515:Plant Leaves / Q000737:chemistry / Q000378:metabolism; D030821:Plants, Genetically Modified / Q000276:immunology*; D011817:Rabbits; D014026:Nicotiana / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D014994:Xylose / Q000032:analysis / Q000276:immunology
chemical_list: C529157:2G12 monoclonal antibody; D000906:Antibodies; D000911:Antibodies, Monoclonal; D052179:Antigens, Plant; D000080908:Broadly Neutralizing Antibodies; D002241:Carbohydrates; D015483:HIV Antibodies; D005643:Fucose; D007073:Immunoglobulin E; D014994:Xylose