Analysis of the O-linked glycan structures of a glycoprotein determines their glycan type and abundance.
O-linked glycan analysis is applicable to all stages of biopharmaceutical development and manufacturing processes including cell line and clone selection, upstream and downstream process development, manufacturing in-process control, batch consistency and release. O-linked glycan analysis is required by ICH Q6B guidelines for biopharmaceutical characterization.
O-linked glycan profiles are obtained by releasing the glycans from the glycoprotein by hydrazinolysis, labeling of the glycans with fluorescence dye, 2-Aminobenzamide (2AB), and separating them by normal phase (NP) or reverse phase (RP) HPLC columns.
In order to obtain full structural analysis, enzyme array of exoglycosidases is applied to the labeled glycans, followed by re-separation on the NP or RP chromatography. The peaks are verified according to their retention time.
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