Production of a monoclonal antibody in plants with a humanized N ?￠????glycosylation pattern

摘要

In recent years, plants have become an attractive alternative for the production of recombinant proteins. However, their inability to perform authentic mammalian N ?￠????glycosylation may cause limitations for the production of therapeutics. A major concern is the presence of ???21,2?￠????xylose and core ???±1,3?￠????fucose residues on complex N ?￠????linked glycans, as these N ?￠????glycan epitopes are immunogenic in mammals. In our attempts towards the humanization of plant N ?￠????glycans, we have generated an Arabidopsis thaliana knockout line that synthesizes complex N ?￠????glycans lacking immunogenic xylose and fucose epitopes. Here, we report the expression of a monoclonal antibody in these glycan?￠????engineered plants that carry a homogeneous mammalian?￠????like complex N ?￠????glycan pattern without ???21,2?￠????xylose and core ???±1,3?￠????fucose. Plant and Chinese hamster ovary (CHO)?￠????derived immunoglobulins (IgGs) exhibited no differences in electrophoretic mobility and enzyme?￠????linked immunosorbent specificity assays. Our results demonstrate the feasibility of a knockout strategy for N ?￠????glycan engineering of plants towards mammalian?￠????like structures, thus providing a significant improvement in the use of plants as an expression platform.