Production of mouse monoclonal antibody with galactose-extended sugar chain by suspension cultured tobacco BY2 cells expressing human beta(1,4)-galactosyltransferase.
Previously, we developed a transgenic tobacco BY2 cell line (GT6) in which glycosylation was modified by expressing human beta(1,4)-galactosyltransferase (betaGalT). In this study, we produced a mouse monoclonal antibody in GT6 cells, and determined the sugar chain structures of plant-produced antibodies. Galactose-extended N-linked glycans comprised 16.7%, and high-mannose-type and complex-type glycans comprised 38.5% and 35.0% of the total number of glycans, respectively. N-linked glycans with the plant-specific sugars beta(1,2)-xylose and alpha(1,3)-fucose comprised 9.8%. The introduction of human betaGalT into suspension cultured tobacco cells resulted in the production of recombinant proteins with galactose-extended glycans and decreased contents of beta(1,2)-xylose and alpha(1,3)-fucose.
Fujiyama
Kazuhito
K
The International Center for Biotechnology, Osaka University, 2-1 Yamada-oka, Suita, Osaka 565-0871, Japan. fujiyama@icb.osaka-u.ac.jp
Furukawa
Aiko
A
Katsura
Atsuya
A
Misaki
Ryo
R
Omasa
Takeshi
T
Seki
Tatsuji
T
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics; D002240:Carbohydrate Sequence; D002460:Cell Line; D005690:Galactose / Q000737:chemistry; D006023:Glycoproteins / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D009237:N-Acetyllactosamine Synthase / Q000096:biosynthesis*; D030821:Plants, Genetically Modified / Q000235:genetics / Q000378:metabolism; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics; D014026:Nicotiana / Q000235:genetics / Q000378:metabolism*
D000911:Antibodies, Monoclonal; D006023:Glycoproteins; D011994:Recombinant Proteins; D009237:N-Acetyllactosamine Synthase; D005690:Galactose