N-Glycosylation of an IgG antibody secreted by Nicotiana tabacum BY-2 cells can be modulated through co-expression of human β-1,4-galactosyltransferase.
Nicotiana tabacum BY-2 suspension cells have several advantages that make them suitable for the production of full-size monoclonal antibodies which can be purified directly from the culture medium. Carbohydrate characterization of an antibody (Lo-BM2) expressed in N. tabacum BY-2 cells showed that the purified Lo-BM2 displays N-glycan homogeneity with a high proportion (>70%) of the complex GnGnXF glycoform. The stable co-expression of a human β-1,4-galactosyltransferase targeted to different Golgi sub-compartments altered Lo-BM2N-glycosylation and resulted in the production of an antibody that exhibited either hybrid structures containing a low abundance of the plant epitopes (α-1,3-fucose and β-1,2-xylose), or a large amount of galactose-extended N-glycan structures. These results demonstrate the suitability of stable N-glycoengineered N. tabacum BY-2 cell lines for the production of human-like antibodies.
Institut des Sciences de la Vie, Université catholique de Louvain, 1348, Louvain-la-Neuve, Belgium. catherine.navarre@uclouvain.be.
Catherine
0000-0002-8413-0878
C
Navarre
Mass Spectrometry Laboratory, University of Liege, 4000, Liège, Belgium.
Nicolas
N
Smargiasso
Institut des Sciences de la Vie, Université catholique de Louvain, 1348, Louvain-la-Neuve, Belgium.
Laurent
L
Duvivier
Institut des Sciences de la Vie, Université catholique de Louvain, 1348, Louvain-la-Neuve, Belgium.
Joseph
J
Nader
Mass Spectrometry Laboratory, University of Liege, 4000, Liège, Belgium.
Johann
J
Far
Mass Spectrometry Laboratory, University of Liege, 4000, Liège, Belgium.
Edwin
E
De Pauw
Institut des Sciences de la Vie, Université catholique de Louvain, 1348, Louvain-la-Neuve, Belgium.
Marc
M
Boutry
D002846:Chromatography, Affinity; D005786:Gene Expression Regulation; D006031:Glycosylation; D006056:Golgi Apparatus / Q000378:metabolism; D006801:Humans; D007074:Immunoglobulin G / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D009237:N-Acetyllactosamine Synthase / Q000235:genetics* / Q000378:metabolism; D030821:Plants, Genetically Modified / Q000235:genetics*; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D014026:Nicotiana / Q000235:genetics* / Q000378:metabolism
D007074:Immunoglobulin G; D011994:Recombinant Proteins; D009237:N-Acetyllactosamine Synthase
Antibody; N-Glycosylation; Plant suspension cells; β-1,4-Galactosyltransferase