Henquet, M.M.HenquetEigenhuijsen, J.J.EigenhuijsenHesselink, T.T.HesselinkSpiegel, H.H.SpiegelSchreuder, M.M.SchreuderDuijn, E. vanE. vanDuijnCordewener, J.J.CordewenerDepicker, A.A.DepickerKrol, A. van derA. van derKrolBosch, D.D.Bosch2022-03-042022-03-042011https://publica.fraunhofer.de/handle/publica/22559510.1007/s11248-010-9475-5ER resident glycoproteins, including ectopically expressed recombinant glycoproteins, carry so-called high-mannose type N-glycans, which can be at different stages of processing. The presence of heterogeneous high-mannose type glycans on ER-retained therapeutic proteins is undesirable for specific therapeutic applications. Previously, we described an Arabidopsisalg3-2 glycosylation mutant in which aberrant Man5GlcNAc2 mannose type N-glycans are transferred to proteins. Here we show that the alg3-2 mutation reduces the N-glycan heterogeneity on ER resident glycoproteins in seeds. We compared the properties of a scFv-Fc, with a KDEL ER retention tag (MBP10) that was expressed in seeds of wild type and alg3-2 plants. N-glycans on these antibodies from mutant seeds were predominantly of the intermediate Man5GlcNAc2 compared to Man8GlcNAc2 and Man7GlcNAc2 isoforms on MBP10 from wild-type seeds. The presence of aberrant N-glycans on MBP10 did not seem to affect MBP10 dimerisa tion nor binding of MBP10 to its antigen. In alg3-2 the fraction of underglycosylated MBP10 protein forms was higher than in wild type. Interestingly, the expression of MBP10 resulted also in underglycosylation of other, endogenous glycoproteins.enN-glycosylationrecombinant antibodyMolecular FarmingArabidopsis540576journal article