Eculizumab (Soliris, Alexion), trastuzumab
(Herceptin, Roche), and pembrolizumab (Keytruda, Merck) were obtained
from their manufacturers. Glycan remodeling was performed by treatment
of trastuzumab with TransGLYCIT (Genovis, Lund, Sweden) according
to the producer specifications with and without the presence of a
fucosidase. For middle-level analysis, enzymatic digestion was performed
by incubation of one unit of the IdeS enzyme (FabRICATOR, Genovis,
Lund, Sweden) per microgram of mAb at 37 °C for 60 min. Prior
to further analysis, the samples were buffer exchanged to 50 mM ammonium
acetate using 10 kDa Vivaspin 500 filters (Sartorius, Göttingen,
Germany) with an end concentration of 2 mg/mL.
(Herceptin, Roche), and pembrolizumab (Keytruda, Merck) were obtained
from their manufacturers. Glycan remodeling was performed by treatment
of trastuzumab with TransGLYCIT (Genovis, Lund, Sweden) according
to the producer specifications with and without the presence of a
fucosidase. For middle-level analysis, enzymatic digestion was performed
by incubation of one unit of the IdeS enzyme (FabRICATOR, Genovis,
Lund, Sweden) per microgram of mAb at 37 °C for 60 min. Prior
to further analysis, the samples were buffer exchanged to 50 mM ammonium
acetate using 10 kDa Vivaspin 500 filters (Sartorius, Göttingen,
Germany) with an end concentration of 2 mg/mL.
Free full text: Click here
