Bioreactor Product Glycosylation Profile Assessment by the A2P2
System
In the case of sample preparation for a glycosylation profile assay, if
the starting material is a cell culture sample, the process includes
affinity protein cleanup, enzymatic release of glycans from the protein,
chemical labeling of the released glycans with fluorescent tags, and
subsequent HILIC chromatography column injection and analysis. As
depicted in Figure 8c , the autosampler receives manually
collected or automatically delivered bioreactor cell-free samples in
vials or through the FTV. The autosampler needle takes the sample and
injects it into the affinity capture column for cell culture matrix
removal. The autosampler needle then delivers PNGase F to the capture
column for on-column glycan release. The released glycans are
transferred to the FTV, mixed with 2-AA, and directed to the 80°C
reaction coil, where the fluorescent labeling reaction takes place. The
fluorescently labeled glycans are then conveyed back to the FTV, ready
for normal-phase chromatographic separation and quantitation.