Principal Scientist Amgen Cambridge, Massachusetts
High molecular weight (HMW) species, a subset of size variants, represent a critical quality attribute for many therapeutic monoclonal antibodies (mAbs). Monitoring HMW species is essential throughout process development and characterization. In this case study, we describe the findings from development of a size exclusion chromatography (SEC) method for in-process sample characterization of a mAb. Multiple SEC columns were evaluated, revealing variable recovery of higher-order aggregates. Notably, column performance stabilized after repeated injections, with the number of injections required varying from column to column. While SEC methods typically use a reference standard (RS) for column conditioning, the RS in this case contained only dimers and failed to resolve the issue. Instead, in-process samples containing higher-order aggregates were necessary to achieve consistent column performance.
Learning Objectives:
Upon completion, participants will be able to identify challenges in HMW recovery across SEC columns and understand the need for appropriate sample-based conditioning to ensure consistent detection of higher-order aggregates.
