FLUOROPHORE-ASSISTED CARBOHYDRATE ELECTROPHORESIS (FACE(R)) FOR QUALITY-CONTROL OF RECOMBINANT-PROTEIN GLYCOSYLATION

A large amount of evidence now exists to show that protein glycosylati on plays a critical role in the pharmacokinetics and bioactivity of ma ny medically important glycoproteins. This realization has led many bi otechnology laboratories to establish in-house analytical protocols th at can be used to monitor the nature and consistency of glycosylation during the manufacture of recombinant glycoprotein therapeutics. This paper describes a relatively simple and highly sensitive method called fluorophore-assisted carbohydrate electrophoresis, or FACE(R), for pe rforming routine carbohydrate analysis. Specifically, we describe a se ries of experimental protocols that can be used to form the framework for in-house validation of the FACE(R) method, The influence of variab les in sample preparation, electrophoresis conditions and gel formulat ions on carbohydrate band quantification and band mobility of asparagi ne-linked oligosaccharide profiles is evaluated.