Recombinant human acid alpha-glucosidase (
rhGAA) from Chinese hamster ovary cells is the only approved treatment for patients with
Pompe disease. In this study, rhGAAs were produced in transgenic rice cell
suspension cultures under eight different conditions; untreated, 5 μM of 2-fluoro-l-fucose (2-FF), 50 μM of 2-FF, 100 μM of 2-FF, 100 μM of 2-FF + 0.5%
Pluronic F-68 (PF-68), 100 μM of 2-FF + 0.05%
Tween 20 (Tw 20), 0.5% PF-68, and 0.05% Tw 20. The N-
glycans of eight rhGAAs were analyzed using ultra-performance liquid chromatography (UPLC) and tandem mass spectrometry. The relative quantity (%) of each
glycan was obtained from the corresponding UPLC peak area per the sum (100%) of individual UPLC peak area. Fifteen N-
glycans, comprising seven core-fucosylated
glycans (71.5%, sum of each relative quantities) that have immunogenicity-inducing potential, three de-core-fucosylated
glycans (15.4%), and five non-core-fucosylated
glycans (13.1%), were characterized with high mass accuracy and
glycan-generated fragment
ions. The increases or decreases of relative quantities of each
glycan from seven rhGAAs were compared with those of untreated control. The percentages of the sum of the relative quantities of core-fucosylated
glycans divided by the sums of those of de-core- (core-
fucose removed) and non-core-fucosylated
glycans were calculated, and the lowest percentage was obtained in 100 μM of 2-FF combined with 0.5% PF-68. These results indicate that the relative quantity of each
glycan of
rhGAA produced in rice cell
suspension cultures is significantly affected by their culture condition. This study performed the comparison of the N-
glycan profiles of rice cell-derived
rhGAA to identify the core-fucosylated
glycans using UPLC and tandem mass spectrometry.