N-linked glycan structures of a mouse monoclonal antibody produced from tobacco BY2 suspension-cultured cells.
cDNA encoding H- and L-chains from a mouse monoclonal antibody was introduced into tobacco BY2 cells, and the resulting sugar chain structures of plant-produced antibodies were analyzed by a combination of HPLC, exoglucosidase digestion and matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry. The glycan structures determined were Man(5-6)GlcNAc2 (22.3%), GlcNAcMan5GlcNAc2 (3.1%), GlcNAcMan3FucXylGlcNAc2 (24.4%), GlcNAcMan3XylGlcNAc2 (17.8%), Man3FucXylGlcNAc2 (24.3%), and Man3XylGlcNAc2 (8.1%). The major glycan structures of the antibodies produced by transgenic suspension-cultured cells contain typical plant bisecting beta(1,2)-xylose and alpha(1,3)-fucose residues, suggesting the posttranslational modification of a recombinant antibody in the late Golgi apparatus.
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
Misaki
Ryo
R
Katsura
Atsuya
A
Tanaka
Tomoko
T
Furukawa
Aiko
A
Omasa
Takeshi
T
Seki
Tatsuji
T
D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry*; D001483:Base Sequence; D002240:Carbohydrate Sequence; D003001:Cloning, Molecular; D017343:Genes, Plant; D005959:Glucosidases / Q000378:metabolism; D006056:Golgi Apparatus / Q000378:metabolism; D007074:Immunoglobulin G / Q000737:chemistry; D051379:Mice; D008969:Molecular Sequence Data; D010944:Plants / Q000737:chemistry / Q000378:metabolism; D011134:Polysaccharides / Q000737:chemistry*; D014026:Nicotiana / Q000378:metabolism*
D000911:Antibodies, Monoclonal; D007074:Immunoglobulin G; D011134:Polysaccharides; D005959:Glucosidases