Production and N-glycan engineering of Varlilumab in Nicotiana benthamiana

N -glycan engineering has dramatically evolved for the development and quality control of recombinant antibodies. Fc region IgG contains two -glycans whose galactose terminals on Fc-glycan have been shown to increase stability CH2 domain improve effector functions. Nicotiana benthamiana become one most attractive production systems therapeutic In this study, Varlilumab, a CD27-targeting monoclonal antibody, was transiently produced in fresh leaves soil-grown hydroponic-grown N. , resulted yield 174 618 µg/gram, respectively. However, wild-type lacked terminal residues its -glycan. Therefore, applied fine-tune antibodies plant platforms. We further co-expressed together with murine β1,4-galactosyltransferase (β1,4-GALT) modify β1,4-linked Gal residue(s) Arabidopsis thaliana β1,3-galactosylatransferase (β1,3-GALT) galactosylation. The co-expression each GALTs successfully modification structures plant-produced IgG. Notably, β1,4-GALT had 42.5% variants having (Gal) at non-reducing terminus 55.3% that plants. Concomitantly, N- glycan profile analysis β1,3-GALT demonstrated there an increased efficiency galactosylation enhancement formation Lewis structure plant-derived Taken together, our findings show first Varlilumab biantennary β1,4-galactosylated structures.