ENHANCING ANALYTICAL PERFORMANCE WITH A BIO-INERT HPLC SYSTEM

Importance of the Topic

The accurate quantitation and high-resolution separation of biotherapeutic analytes is fundamental to drug development, quality control and regulatory compliance. Traditional stainless-steel HPLC systems can promote undesired analyte–surface interactions, leading to adsorption, reduced recovery and compromised reproducibility.

Objectives and Study Overview

This evaluation compared the legacy steel-based HPLC platform with the new Alliance iS Bio HPLC System featuring MaxPeak High Performance Surfaces (HPS) technology. The primary goals were to assess improvements in resolution, recovery and signal-to-noise ratio for diverse biopharmaceutical molecules.

Methodology

Established methods employing ion-pairing reversed-phase LC (IP-RPLC), size-exclusion chromatography (SEC) and standard reversed-phase LC were transferred from the legacy system to the Alliance iS Bio platform. Analytes included

• MassPREP Oligonucleotide Standard (15–35 mer OST)
• GEM91 phosphorothioate oligonucleotide
• USP monoclonal antibody reference standard
• Glucagon-like peptide-1 (GLP-1) peptides

Used Instrumentation

• Alliance iS Bio HPLC System with MaxPeak HPS coatings
• XBridge Premier Oligonucleotide BEH C18 column (2.5 µm, 130 Å, 4.6 × 100 mm)
• XBridge BEH C18 columns (2.5 µm & 5 µm, 130 Å, 4.6 × 100 mm)
• Legacy stainless-steel flow path and columns for comparison
• UV detector at 260 nm

Results and Discussion

Key observations included

• 30% increase in peak height for 15–35 mer oligonucleotides on the HPS-coated system versus legacy steel
• Approximately 40% improvement in signal-to-noise ratio for trace impurities in GEM91 analyses
• Consistent retention times and enhanced reproducibility for monoclonal antibody reference standards
• Maximized recovery of GLP-1 peptides and elimination of lengthy column passivation procedures

Benefits and Practical Applications

The bio-inert HPLC system reduces method development time by removing surface adsorption concerns, accelerates throughput and delivers robust quantitation for critical biopharmaceutical assays in research, QC and regulated environments.

Future Trends and Applications

Continued advances may include specialized surface chemistries optimized for direct mass spectrometry coupling, integration with microfluidic platforms for high-throughput screening and extension of bio-inert technologies to capillary and preparative chromatography scales.

Conclusion

The Alliance iS Bio HPLC System with MaxPeak HPS technology effectively mitigates analyte–surface interactions, providing improved sensitivity, recovery and reproducibility for a wide range of biotherapeutic analyses.

Reference

Bigos P, Du X, Han D, Birdsall R, Hoehnel A, Nyholm K. Enhancing analytical performance with a bio-inert HPLC system Waters Corporation