Change in glycosylation pattern with extension of endoplasmic reticulum retention signal sequence of mouse antibody produced by suspension-cultured tobacco BY2 cells.
The production of antibodies using plants as bioreactors has been realized. Because sugar chain structures on recombinant proteins are a cause of concern, remodeling technology is highly promising. Localizing recombinant proteins in the endoplasmic reticulum (ER) affects the glycosylation pattern in transgenic plants. Previously, a mouse antibody produced by suspension-cultured tobacco BY2 cells has sugar chains with possible glycoepitopes as the predominant structures. In this study, we extended the Lys-Asp-Glu-Leu (KDEL) ER retention signal sequence over the heavy (H) and light (L) chains of the antibody and expressed the altered antibody in tobacco BY2 cells to study the effect of the KDEL sequence on glycosylation. For the antibody with the KDEL-extended H-chains, glycans with beta(1,2)-xylose or alpha(1,3)-fucose residues accounted for 49% of the total glycans. Meanwhile, for the antibody with the KDEL-extended H- and L-chains, glycans with xylose or fucose accounted for 38% of the total glycans. Although the addition of an ER retention signal shifted the dominant glycan structures of the KDEL-extended antibody to high-mannose-type structures, some of the antibodies escaped the retrieval system during intracellular traffic and were then modified by xylosylation or fucosylation.
Fujiyama
Kazuhito
K
International Center for Biotechnology, Osaka University, 2-1 Yamada-oka, Suita, Osaka 565-0871, Japan. fujiyama@icb.osaka-u.ac.jp
Misaki
Ryo
R
Sakai
Yohei
Y
Omasa
Takeshi
T
Seki
Tatsuji
T
D000818:Animals; D000906:Antibodies / Q000302:isolation & purification; D000917:Antibody Formation; D001483:Base Sequence; D002460:Cell Line; D002851:Chromatography, High Pressure Liquid; D017931:DNA Primers; D004721:Endoplasmic Reticulum / Q000378:metabolism*; D006031:Glycosylation; D051379:Mice; D030821:Plants, Genetically Modified; D016133:Polymerase Chain Reaction; D021382:Protein Sorting Signals; D014026:Nicotiana / Q000166:cytology*
D000906:Antibodies; D017931:DNA Primers; D021382:Protein Sorting Signals