ADVANCING ANALYTICAL PERFORMANCE FOR BIOTHERAPEUTICS WITH A BIO-INERT HPLC SYSTEM

Significance of the topic

Biotherapeutic analytics rely on high performance liquid chromatography to monitor drug quality, but standard stainless steel flow paths can cause adsorption of analytes, leading to loss of signal and poor recovery. Developing bio-inert HPLC systems is critical for accurate, reliable analysis of modern biotherapeutic modalities.

Objectives and overview of the study

This study compares the Alliance iS Bio HPLC System featuring MaxPeak High Performance Surfaces with a legacy stainless steel HPLC platform. The goal is to assess improvements in resolution, signal-to-noise ratio and recovery for oligonucleotides, monoclonal antibodies and peptide therapeutics.

Methodology

Established ion-pairing reversed phase chromatography, size exclusion chromatography and reversed phase liquid chromatography methods were transferred from legacy HPLC to the bio-inert system. Chromatographic conditions included XBridge BEH C18, BioSuite Diol and Premier Protein SEC columns, as well as Peptide CSH C18 columns, various salt concentrations and gradient elution profiles to challenge system performance.

Results and discussion

Analysis of a fully thiolated oligonucleotide showed a 40% increase in signal-to-noise for trace impurities on the bio-inert system. Size exclusion chromatography of a monoclonal antibody demonstrated wider salt tolerance and lower baseline noise, enhancing detection of high and low molecular weight species. A panel of GLP-1 peptides exhibited sharper peak shapes and improved integration accuracy due to reduced system dispersion.

Benefits and practical applications

The bio-inert HPLC system delivers higher sensitivity, resolution and recovery across multiple biotherapeutic classes. This enables robust quality control workflows in manufacturing and research laboratories, supporting routine analysis of novel modalities and traditional biologics.

Future trends and potential applications

Further developments may integrate this bio-inert platform with mass spectrometry and advanced detectors, extend its use to cell and gene therapy products, and enhance automation for high throughput biopharma environments.

Conclusion

Adoption of bio-inert flow paths significantly improves analytical performance for biotherapeutics, offering consistent precision and accuracy compared to legacy systems. The Alliance iS Bio HPLC System represents a modern solution for demanding QC applications.

Used Instrumentation

• Alliance iS Bio HPLC System with MaxPeak High Performance Surfaces Technology
• Legacy stainless steel HPLC System
• XBridge BEH C18 Column 5 um 130 Å 4.6 x 100 mm
• BioSuite Diol Column 250 Å 5 um 7.8 x 300 mm
• Premier Protein SEC Column 250 Å 2.5 um 7.8 x 300 mm
• Peptide CSH C18 Column 130 Å 2.5 um 4.6 x 150 mm

References

• Du X Birdsall RE Bigos P Han D Nyholm K Deploying the Alliance iS Bio HPLC System as a modern HPLC for biopharmaceutical analysis in QC Environments Waters Application Note 720008288EN
• Bigos P Birdsall RE Nyholm K Evaluating the Ruggedness of the Alliance iS Bio HPLC System for SEC Separations Waters Application Note 720008427EN
• Han D Birdsall RE Nyholm K Leveraging the Alliance iS Bio HPLC System as a Modern HPLC for Peptide Drug Substances Analysis in QC Environments Waters Application Note 720008345EN