High throughput and rapid size exclusion chromatography (SEC) methods can be useful to support the development of biotherapeutic processes and formulations. Additionally, a fast SEC method will be more efficient for real-time purity analyzes to support a production process. In these applications, SEC can be used to monitor self-associated or aggregated protein impurities or protein fragmentation that may impact product safety or efficacy.
In a previous application note, it was demonstrated that efficient separations and extended column lifetimes could be achieved using mobile phases at physiological pH (7.4) and ionic strength using the columns Waters™ MaxPeak Premier Protein SEC. Therefore, the performance and column life evaluation of the UPLC and HPLC versions of these recently introduced columns were evaluated while operating near their maximum specified pressures, resulting in run times of 2.1 to 3.0 minutes.
For this study, the lifetimes of one Waters ACQUITY™ Premier Protein SEC column (250 Å, 1.7 µm, 4.6 x 150 mm) and two XBridge™ Premier Protein SEC columns (250 Å, 2.5 µm) of dimensions 4.6 x 150 mm and 7.8 x 150 mm were successfully evaluated during 500 analyses. Separation performance was assessed for self-associated and selected fragmented size variants of four marketed monoclonal antibody biosimilars.
- Fast SEC analyzes (2.1 to 3.0 minutes), SEC analyzes of mAb size variants on UPLC and HPLC systems
- Column lifetimes greater than 500 runs near specified maximum column pressures
- Reproducible determinations of HMWS and LMWS impurity levels throughout the life study