Monitoring Glycosylation of Therapeutic Glycoproteins for Consistency Using Highly Fluorescent Anthranilic Acid

Analysis of carbohydrates and glycoconjugates by matrix‐assisted laser desorption/ionization mass spectrometry: An update for 2003–2004

“…Advantages of analytical derivatization have been emphasized by Rosenfeld (2003) with, among others, examples from the reductive amination of carbohydrates for improving the interpretation of fragmentation spectra and for modifying behavior in chromatographic systems. MALDI is frequently used to monitor reducing-terminal labeling, used to attach fluorophores or chromophores to carbohydrates for chromatographic detection as illustrated in a recent study of glycans from the Tamm-Horsfall glycoprotein (Rohfritsch et al, 2004) and the use of 2-AAderivatives to monitor glycosylation of therapeutic glycoproteins (Dhume et al, 2002). Benzylamine (2/9) derivatization was used by mainly for ESI-MS/MS analysis of small and N-linked glycans.…”

The Chemistry of Anthranilic Acid

“…The fluorescent properties of AA can be used for bioanalytical purposes, e.g. monitoring of glycosylation of proteins [4]. pK a Medium 6.53 [ Like other amino acids AA is amphoteric.…”

Development and Application of a Robust N‐Glycan Profiling Method for Heightened Characterization of Monoclonal Antibodies and Related Glycoproteins

“…Characterization and routine analysis of N-glycosylation patterns in mAb therapeutics is important for the assessment of product quality and consistency of manufacture in the development and commercialization of protein pharmaceuticals. 1,2,[8][9][10][11][12][13] Commonly, the N-linked oligosaccharides are characterized and monitored through enzymatic release from the mAb with an endoglycosidase with subsequent separation of the individual native or fluorescently labeled structures by chromatographic [14][15][16][17][18][19] or electrophoretic methods. 11,20,21 The resulting distribution of peaks serves as an "N-glycan fingerprint" where particular carbohydrate structures are indicated by peaks at specific retention (or migration) times and their relative abundances are reflected by the peak areas.…”