Improving Chromatographic Separations Of Biopharmaceuticals With MaxPeak High Performance Surfaces (HPS) Technology

Importance of the topic

Biopharmaceuticals represent the fastest-growing sector in the pharmaceutical industry and include large molecules such as monoclonal antibodies, peptides, nucleotides, glycans, and lipid nanoparticles. Their complex structures and electron-rich functional groups often interact nonspecifically with metal surfaces in standard stainless-steel chromatography hardware, leading to poor reproducibility, peak distortion, and loss of sensitivity. These challenges demand a comprehensive solution to enhance chromatographic performance, improve confidence in analytical results, and accelerate biotherapeutic discovery, development, and manufacturing.

Study objectives and overview

This expert summary reviews the implementation of Waters’ MaxPeak™ High Performance Surfaces (HPS) Technology across a series of application notes, demonstrating its impact on liquid chromatography (LC) and LC-mass spectrometry (MS) workflows for biopharmaceutical analytics. The compiled studies cover intact protein size-exclusion chromatography (SEC), peptide mapping and quality attribute monitoring, nucleotide bioanalysis, glycan profiling, lipid nanoparticle characterization, mRNA poly(A) tail measurement, and cell culture media monitoring. Each application highlights improvements in resolution, sensitivity, accuracy, and robustness enabled by HPS technology in both columns and instrument flow paths.

Methods and instrumentation

MaxPeak HPS employs a hybrid organic/inorganic barrier coating on metal surfaces to suppress ionic and hydrophobic adsorption. It has been integrated into:

• Column hardware (ACQUITY Premier and XBridge Premier Columns, BioResolve SEC and SCX Columns, ACQUITY Premier Glycan BEH C18 AX, Amide Columns, Oligonucleotide C18 Columns).
• Instrument platforms (ACQUITY Premier UPLC System, Arc Premier UHPLC System, ACQUITY QDa Mass Detector, BioAccord LC-MS System, Xevo G3 QTof MS).
• Informatics solutions (Empower Software, waters_connect, UNIFI, Column Calculator).

Analytical techniques included reversed-phase MS (RP-MS), ion-exchange UV-MS (IEX-UV-MS), peptide mapping, multiple attribute method (MAM), hydrophilic interaction chromatography (HILIC-FLR/MS), mixed-mode glycan separations, ion-pair reversed-phase for oligonucleotides, evaporative light scattering (ELSD), and size-exclusion chromatography with UV detection.

Main results and discussion

Across all modalities, MaxPeak HPS Technology delivered:

• Reduced metal-induced secondary interactions, shown by up to 200% lower nonspecific binding in SEC of intact proteins and decreased peak tailing in cation-exchange chromatography.
• Enhanced recovery of acidic and phosphorylated peptides, yielding up to 34-fold increases in MS response and four-fold gains in peak area compared to conventional hardware.
• Improved reproducibility of peptide mapping retention times (standard deviation <1 second) and lower RSDs in peptide quality attribute monitoring over three orders of magnitude in dynamic range.
• Superior oligonucleotide chromatographic performance without system passivation, delivering tenfold better recovery and extended dynamic range in impurity quantification.
• Orthogonal lipid nanoparticle analysis by ELSD/PDA with complementary mass detection, achieving picogram sensitivity for key lipid components.
• A rapid SEC-UV method for poly(A) tail length determination of mRNA, exploiting RNase T1 digestion and calibration curves from oligoribonucleotide standards.
• Improved recovery and peak shape of highly-sialylated and phosphorylated glycans in HILIC and mixed-mode separations, enhancing FLR/MS sensitivity and enabling automated glycan assignments.
• High-throughput cell culture media nutrient and metabolite monitoring using BioAccord LC-MS, integrating trend plots, unknown screening, and multivariate analysis for microbial and mammalian bioprocess development.
• Successful migration of legacy amino acid analysis with pre-column derivatization to the ACQUITY Premier Binary System, preserving resolution, tailing, linearity, and precision.

Benefits and practical applications

The application of MaxPeak HPS Technology offers significant advantages for biopharmaceutical laboratories:

• Reliable size variant and charge variant assessments without high-salt or organic mobile phases.
• Streamlined biosimilar comparability studies at intact and subunit levels using ACQUITY Premier Columns and Xevo G3 QTof.
• Automated method scaling between UPLC and UHPLC systems via Column Calculator for seamless downstream migration.
• Reduction of assay failures in peptide mapping and MAM workflows through enhanced hardware inertness.
• Integrated LC-MS platform (BioAccord) for comprehensive analysis of peptides, glycans, oligonucleotides, lipids, and metabolites.
• Compliance-ready informatics for GxP and non-GxP environments, supporting accelerated method development and routine QC testing.

Future trends and possibilities

Looking ahead, the universal application of MaxPeak HPS surfaces can be extended to emerging modalities such as lipid nanoparticle-based gene therapies and next-generation mRNA vaccines. Integration with artificial intelligence algorithms and machine learning may enable predictive method development and real-time monitoring of bioprocess attributes. Further miniaturization and high-throughput capabilities will support continuous manufacturing, single-cell proteomics, and personalized therapeutic analytics.

Conclusion

MaxPeak High Performance Surfaces Technology establishes a new benchmark for chromatographic inertness, delivering improved resolution, sensitivity, and reproducibility across a broad spectrum of biopharmaceutical analyses. By eliminating nonspecific adsorption in both column and instrument hardware, this holistic solution empowers scientists to achieve reliable critical quality attribute measurements, accelerate workflow development, and maintain robust performance in discovery, development, and manufacturing settings.