Module 9 2024

03/09/2024

Purity – Intact Example Method: CE-SDS

Non reduced

Covalent Aggregat e

Heavy Chain

Light Chain

Reduced

Aglycosyl Heavy Chain

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Purity – Glycosylation Example Method: NP-HPLC

- Glycosylation plays a key role in effector function, PK, and is an indicator of process consistency and control - PNGase F used to release N-linked glycans from protein. Oligosaccharides are labelled with the fluorophore 2 aminobenzamide (2-AB) used for identification against standards. - Orthogonal methods – Intact glycoproteins (ESI-MS), glycopeptides (LC-MS, MALDI), released glycans (underivatized – MALDI-TOF, HPAE-PAD) and monosaccharide analysis.

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