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Engineering mammalian cells to produce plant-specific N-glycosylation on proteins.
Glycobiology ( IF 4.3 ) Pub Date : 2020-02-10 , DOI: 10.1093/glycob/cwaa009 Joachim Steen Larsen 1, 2 , Richard Torbjörn Gustav Karlsson 2 , Weihua Tian 2 , Morten Alder Schulz 2 , Annemarie Matthes 1 , Henrik Clausen 2 , Bent Larsen Petersen 1, 2 , Zhang Yang 2
Glycobiology ( IF 4.3 ) Pub Date : 2020-02-10 , DOI: 10.1093/glycob/cwaa009 Joachim Steen Larsen 1, 2 , Richard Torbjörn Gustav Karlsson 2 , Weihua Tian 2 , Morten Alder Schulz 2 , Annemarie Matthes 1 , Henrik Clausen 2 , Bent Larsen Petersen 1, 2 , Zhang Yang 2
Affiliation
Protein N-glycosylation is an essential and highly conserved posttranslational modification found in all eukaryotic cells. Yeast, plants and mammalian cells, however, produce N-glycans with distinct structural features. These species-specific features not only pose challenges in selecting host cells for production of recombinant therapeutics for human medical use but also provide opportunities to explore and utilize species-specific glycosylation in design of vaccines. Here, we used reverse cross-species engineering to stably introduce plant core α3fucose (α3Fuc) and β2xylose (β2Xyl) N-glycosylation epitopes in the mammalian Chinese hamster ovary (CHO) cell line. We used directed knockin of plant core fucosylation and xylosylation genes (AtFucTA/AtFucTB and AtXylT) and targeted knockout of endogenous genes for core fucosylation (fut8) and elongation (B4galt1), for establishing CHO cells with plant N-glycosylation capacities. The engineering was evaluated through coexpression of two human therapeutic N-glycoproteins, erythropoietin (EPO) and an immunoglobulin G (IgG) antibody. Full conversion to the plant-type α3Fuc/β2Xyl bi-antennary agalactosylated N-glycosylation (G0FX) was demonstrated for the IgG1 produced in CHO cells. These results demonstrate that N-glycosylation in mammalian cells is amenable for extensive cross-kingdom engineering and that engineered CHO cells may be used to produce glycoproteins with plant glycosylation.
更新日期：2020-02-10
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