Evaluating the Ruggedness of the Alliance™ iS Bio HPLC System for SEC Separations

Biopharmaceutical companies are often progressing several monoclonal antibody (mAb) drug candidates to mitigate the risks associated with drug development. This strategic approach, aimed at increasing the likelihood of success, is accompanied by rugged technologies to improve efficiency and effectiveness of a labs operation to progress the most promising drug candidate forward. Specifically, in quality control (QC) environments, the implementation of robust methods is critical to ensure precision, accuracy, and reliability of results to make informed decisions throughout the manufacturing process. A key component of these robust methodologies is the selection of instrumentation and columns for the appropriate analytical method. For example, in size-exclusion chromatography (SEC), factors such as the type of stationary phase, column dimensions, instrument dispersion, and instrument ionic strength and pH tolerance should be considered. Recently, Waters has developed the Alliance iS Bio HPLC System for robust analysis of biopharmaceutical applications. This next-generation HPLC system, with biocompatible and exceedingly inert construction, contains MaxPeak™ High Performance Surfaces (HPS) Technology to reduce analysis variability. This application note will assess the robustness benefits associated with this surface technology for SEC separations.

In this study, the Alliance iS Bio HPLC System with an XBridge™ Premier Protein SEC Column (250 Å, 2.5 µm, 7.8 x 300 mm) was evaluated for method robustness and compared to a legacy stainless-steel system and column (250 Å, 5 µm, 7.8 mm x 300 mm). A widely adopted compendial SEC method outlined in the United States Pharmacopeia (USP) General Chapter <129> was adjusted by altering method parameters such as pH, injection volume, sample concentration, salt concentration, and salt type.1 A comparative analysis was performed to evaluate the performance of each instrument configuration when method parameters were adjusted.

Benefits
- The Alliance iS Bio HPLC System coupled with an XBridge Premier Protein SEC Column showed improved tolerance to low ionic strength mobile phases when compared to a stainless-steel system and column