Unveiling the Accurate Site-Specific <i>N</i>- and <i>O</i>-Glycosylation of Hyperglycosylated Erythropoietin Drugs by an Integrated Approach.

ABSTRACT: Hyperglycosylated proteins with a high sialic acid content show great promise in the development of long-acting biotherapeutics. However, their structural complexity and heterogeneity pose significant challenges to traditional analytical methods, which often fail to provide comprehensive glycan information across all glycosylation sites, leading to ambiguities in characterization. Despite the fact that long-acting hyperglycosylated erythropoietin (hyperEPO) has been available on the market for over two decades, its site-specific glycosylation profile remains ambiguous due to closely spaced glycosylation sites and large glycans that contain labile sialic acids substituents. Here, using hyperEPO as an example, we developed an integrated workflow that incorporates an experimentally cross-vali

SUBMITTER: Wu S 

PROVIDER: S-EPMC12619957 | biostudies-literature | 2025 Jul

REPOSITORIES: biostudies-literature