Application of Analytical Methods to Characterize Protein Aggregation for a Biopharmaceutical Drug Program

摘要

The aggregate studied here is a mixture of species, from dimers up to large multimers (18-mer). Agg #2 has a larger mass aggregate than Agg #3. SDS-PAGE gels, SEC, SEC-SLS, AUC, Intact HPLC-MS The aggregated fractions contain a mix of reducible and nonreducible aggregates. SDS-PAGE gels, Intact HPLC-MS Disulfide bond scrambling is the source of some, but not all, of the aggregate. Free sulfhydryl analyses, Disulfide bond mapping Aggregation reduces activity where Drug > Agg #3 > Agg #2 cAMP assay.