{"database":"biostudies-literature","file_versions":[],"scores":null,"additional":{"submitter":["Spanov B"],"funding":["European Commission"],"pagination":["1505-1513"],"full_dataset_link":["https://www.ebi.ac.uk/biostudies/studies/S-EPMC9974696"],"repository":["biostudies-literature"],"omics_type":["Unknown"],"volume":["415(8)"],"pubmed_abstract":["Trastuzumab is known to be heterogeneous in terms of charge. Stressing trastuzumab under physiological conditions (pH 7.4 and 37 °C) increases charge heterogeneity further. Separation of charge variants of stressed trastuzumab at the intact protein level is challenging due to increasing complexity making it difficult to obtain pure charge variants for further characterization. Here we report an approach for revealing charge heterogeneity of stressed trastuzumab at the subunit level by pH gradient cation-exchange chromatography. Trastuzumab subunits were generated after limited proteolytic cleavage with papain, IdeS, and GingisKHAN®. The basic pI of Fab and F(ab)<sub>2</sub> fragments allowed to use the same pH gradient for intact protein and subunit level analysis. Baseline separation of F"],"journal":["Analytical and bioanalytical chemistry"],"pubmed_title":["Revealing charge heterogeneity of stressed trastuzumab at the subunit level."],"pmcid":["PMC9974696"],"funding_grant_id":["H2020 MSCA-ITN 2017 \"Analytics for Biologics\"","H2020 MSCA-ITN 2017 “Analytics for Biologics”"],"pubmed_authors":["Spanov B","Baartmans B","Olaleye O","Bischoff R","Govorukhina N","van de Merbel NC","Nicolardi S","Wuhrer M"],"additional_accession":[]},"is_claimable":false,"name":"Revealing charge heterogeneity of stressed trastuzumab at the subunit level.","description":"Trastuzumab is known to be heterogeneous in terms of charge. Stressing trastuzumab under physiological conditions (pH 7.4 and 37 °C) increases charge heterogeneity further. Separation of charge variants of stressed trastuzumab at the intact protein level is challenging due to increasing complexity making it difficult to obtain pure charge variants for further characterization. Here we report an approach for revealing charge heterogeneity of stressed trastuzumab at the subunit level by pH gradient cation-exchange chromatography. Trastuzumab subunits were generated after limited proteolytic cleavage with papain, IdeS, and GingisKHAN®. The basic pI of Fab and F(ab)<sub>2</sub> fragments allowed to use the same pH gradient for intact protein and subunit level analysis. Baseline separation of F","dates":{"release":"2023-01-01T00:00:00Z","publication":"2023 Mar","modification":"2025-04-22T10:03:51.605Z","creation":"2025-04-05T23:23:24.436Z"},"accession":"S-EPMC9974696","cross_references":{"pubmed":["36693954"],"doi":["10.1007/s00216-023-04547-4"]}}