Evaluating the Waters ACQUITY Premier System as a Flexible LC Platform That Can Be Broadly Deployed in Biopharmaceutical Labs

Importance of the Topic

Biopharmaceutical laboratories require versatile liquid chromatography platforms capable of handling a range of techniques under diverse conditions. The need for high throughput, reliable method transfer and minimal sample loss is critical in the development and manufacturing of complex biomolecules. Demonstrating broad compatibility with high-ionic strength mobile phases further expands the utility of a single LC system across multiple workflows, reducing instrument footprint and simplifying training and maintenance.

Study Objectives and Overview

This study assessed the Waters ACQUITY Premier System equipped with MaxPeak High Performance Surfaces (HPS) Technology as a unified platform for orthogonal LC methods commonly used in biopharmaceutical analysis. Three representative techniques were selected: ion-exchange chromatography (IEX), size-exclusion chromatography (SEC) and hydrophobic interaction chromatography (HIC). Performance on the Premier System was compared against a conventional ACQUITY UPLC System to determine compatibility, reproducibility and the potential for method transfer.

Methodology

For each technique, established biotherapeutic standards were analyzed under industry-typical conditions. IEX separations of a monoclonal antibody (cetuximab) were carried out using a salt gradient in HEPES buffer. SEC analyses of a mAb size variant standard employed isocratic phosphate-chloride mobile phase. HIC experiments on an antibody drug-conjugate surrogate used a high-ionic strength ammonium sulfate gradient. Triplicate runs on both systems enabled statistical comparison of retention times, resolving power (peak/valley ratio) and peak area reproducibility.

Instrumentation Used

• Waters ACQUITY Premier System with MaxPeak HPS Technology
• Waters ACQUITY UPLC System (conventional baseline platform)
• BioResolve SCX mAb Column for IEX
• ACQUITY Protein BEH SEC Column for size-exclusion separations
• Protein-Pak Hi Res HIC Column for hydrophobic interaction chromatog­raphy

Key Results and Discussion

In IEX separations, the Premier System matched the conventional UPLC in retention times, resolving power and peak capacity, demonstrating statistical equivalence across charge variants. For SEC, overall size profiles were equivalent, and the Premier System showed a 34% improvement in peak-to-valley ratio for a clipped fragment, indicating reduced surface adsorption. HIC analysis yielded consistent drug-antibody ratio profiles between platforms, with no significant differences in individual peak areas or total DAR values, confirming robustness under high-ionic strength gradients.

Benefits and Practical Applications

• Single-platform deployment for IEX, SEC and HIC simplifies lab workflows
• High-performance surfaces minimize analyte loss in demanding conditions
• Comparable results enable seamless transfer of legacy LC methods
• Reduced instrument footprint and training burden across multiple techniques

Future Trends and Potential Applications

Expansion of universal LC platforms will support integrated multi-technique workflows, combining chromatography with mass spectrometry and automated sample preparation. Emerging therapeutic modalities, such as bispecific antibodies and cell- and gene-therapy products, will benefit from high-throughput, surface-optimized systems. Continued innovation in stationary phases and surface coatings may further reduce sample adsorption and expand compatibility with novel mobile phases.

Conclusion

The Waters ACQUITY Premier System with MaxPeak HPS Technology delivers reliable, high-quality performance across ion-exchange, size-exclusion and hydrophobic interaction chromatography methods. Its compatibility with high-ionic strength mobile phases and equivalence to conventional platforms streamline method transfer and support efficient analytical operations in biotherapeutic development and manufacturing.

References

• Birdsall R E et al. Increasing Chromatographic Performance of Acidic Peptides in RPLC-MS-Based Assays with ACQUITY Premier Featuring MaxPeak HPS Technology. Waters Application Note 720007003, 2020.
• Koshel B M et al. Improving Recovery and Quantitation of Oligonucleotide Impurities Using ACQUITY Premier with MaxPeak HPS Technology. Waters Application Note 720007238, 2021.
• Kellett J et al. Increasing Recovery of Phosphorylated Peptides Using ACQUITY Premier Technology Featuring MaxPeak High Performance Surfaces. Waters Application Note 720007198, 2021.