HPLC Analysis of Procainamide-Labeled Cetuximab Glycans on BIOshell Glycan using HILIC-FLR
Materials
analytical column
CONDITIONS
column
BIOshell Glycan, 15 cm x 2.1 mm I.D., 2.7 μm particles (55094-U)
mobile phase
[A] 50 mM ammonium formate, pH 4.4 (50 mM ammonium hydroxide, adjusted to pH 4.4 with formic acid); [B] acetonitrile
gradient
75 to 59% B in 75 min
flow rate
0.3 mL/min
column temp.
58 °C
detector
FLR, ex 308 nm, em 359 nm
injection
10 μL
sample
Cetuximab
Description
Analysis Note
Cetuximab is a chimeric mouse-human IgG1 monoclonal antibody against the epidermal growth factor receptor. It is used to treat head and neck as well as colorectal cancers. The antibody is N-glycosylated both in the Fc and Fab regions, which have been shown to impact safety and quality of the drug. Thus, understanding glycosylation patterns is exceptionally important. As shown in this application report, the BIOshell Glycan column is able to elucidate the complex glycosylation of this biotherapeutic, allowing a better understanding of the drug′s efficacy.
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