Production of a monoclonal antibody in plants with a humanized N-glycosylation pattern

• Published in: Plant biotechnology journal 2007-09, Vol.5 (5), p.657-663
• Main Authors: Schähs, Matthias, Strasser, Richard, Stadlmann, Johannes, Kunert, Renate, Rademacher, Thomas, Steinkellner, Herta
• Format: Article
• Language: English
• Subjects: 2-xylose and core α1; 3-fucose; Animals; antibodies; Antibodies, Monoclonal - genetics; Antibodies, Monoclonal - isolation & purification; Antibodies, Monoclonal - metabolism; Arabidopsis - genetics; Arabidopsis - metabolism; CHO Cells; Cricetinae; Cricetulus; Enzyme-Linked Immunosorbent Assay; Epitopes - chemistry; Epitopes - immunology; Gene Deletion; Glycosylation; Humans; Immunoglobulin G - immunology; Immunoglobulin G - isolation & purification; Immunoglobulin G - metabolism; plant glycan engineering; Plant Leaves - genetics; Plant Leaves - metabolism; Plants, Genetically Modified; Polysaccharides - chemistry; Polysaccharides - immunology; Polysaccharides - metabolism; recombinant proteins; Recombinant Proteins - immunology; Recombinant Proteins - isolation & purification; Recombinant Proteins - metabolism; β1,2-xylose and core α1,3-fucose
• ISSN: 1467-7644; 1467-7652
• DOI: 10.1111/j.1467-7652.2007.00273.x

In recent years, plants have become an attractive alternative for the production of recombinant proteins. However, their inability to perform authentic mammalian N-glycosylation may cause limitations for the production of therapeutics. A major concern is the presence of β1,2-xylose and core α1,3-fucose residues on complex N-linked glycans, as these N-glycan epitopes are immunogenic in mammals. In our attempts towards the humanization of plant N-glycans, we have generated an Arabidopsis thaliana knockout line that synthesizes complex N-glycans lacking immunogenic xylose and fucose epitopes. Here, we report the expression of a monoclonal antibody in these glycan-engineered plants that carry a homogeneous mammalian-like complex N-glycan pattern without β1,2-xylose and core α1,3-fucose. Plant and Chinese hamster ovary (CHO)-derived immunoglobulins (IgGs) exhibited no differences in electrophoretic mobility and enzyme-linked immunosorbent specificity assays. Our results demonstrate the feasibility of a knockout strategy for N-glycan engineering of plants towards mammalian-like structures, thus providing a significant improvement in the use of plants as an expression platform.