Method Scaling from ACQUITY™ Premier to Arc™ Premier System

Importance of the Topic

The ability to transfer chromatographic methods reliably between different liquid chromatography systems is critical in biopharmaceutical development and manufacturing workflows. Ensuring consistent separation selectivity and reproducibility reduces development time and resource consumption, supporting high quality control standards.

Objectives and Study Overview

This application note demonstrates the scaling of a peptide mapping method for infliximab from the ACQUITY Premier UPLC System to the Arc Premier UHPLC System using Waters Columns Calculator. Key performance metrics including retention time precision, peak area consistency and peak tailing are evaluated to confirm method equivalency across platforms.

Methodology and Instrumentation

The peptide mapping protocol involved reduction, alkylation and tryptic digestion of infliximab followed by separation under gradient elution. Method scaling was performed with Waters Columns Calculator using system dwell volumes and column dimensions. Instruments employed:

• ACQUITY Premier UPLC System with Premier Peptide CSH C18 column
• Arc Premier UHPLC System with XSelect Premier Peptide CSH C18 column
• QuanRecovery Vials with MaxPeak High Performance Surfaces
• ACQUITY QDa Mass Detector for inline mass detection
• Empower 3 Chromatography Data Software

Key Results and Discussion

Scaled methods preserved separation selectivity with relative retention time shifts below 0.02 and retention time precision below 0.01 minutes. Peak area reproducibility remained within 5 percent RSD under both scaled and custom flow scenarios. MaxPeak High Performance Surfaces significantly reduced peptide peak tailing factors by over 60 percent for acidic peptides, enabling detection and quantitation of low abundance deamidated variants. Inline MS data confirmed consistent mass response across methods.

Benefits and Practical Applications

Method scaling via Waters Columns Calculator:

• Streamlines method transfer across laboratories
• Maintains chromatographic selectivity and sensitivity
• Enables high throughput by custom flow adjustments
• Enhances assay robustness through reduced analyte adsorption

Future Trends and Applications

Advancements in automated method scaling and bioinert surface technologies will support increasing throughput demands, integrate with LCMS workflows more seamlessly and may leverage artificial intelligence for predictive method optimization. Adoption of high performance surfaces is poised to improve trace impurity analysis in next generation biotherapeutics.

Conclusion

The Arc Premier System equipped with MaxPeak High Performance Surfaces and aided by Waters Columns Calculator delivers reproducible peptide mapping equivalent to established UPLC platforms. This approach facilitates efficient method migration, robust assay performance and reliable LCUV and LCMS workflows in biopharmaceutical development and manufacturing.

Reference

1 Robert E Birdsall et al Migration of an HILIC Method for Released Glycan Analysis from the ACQUITY UPLC H Class Bio System to the Arc Premier System Waters Application Note 720007389 2021 Sep
2 Kellen DeLaney et al Improving Peptide Mapping Studies Through Reproducible Performance Using the ACQUITY Premier UPLC System Waters Application Note 720007593 2022 Apr
3 Pawel Bigos et al Obtaining Equivalent IEX Chromatographic Performance Through Automated Method Scaling Using Waters Columns Calculator Waters Application Note 720007807 2022 Dec