ACQUITY UPLC H-Class Bio System
Others | 2011 | WatersInstrumentation
Accurate and rapid characterization of biological macromolecules such as proteins, peptides, nucleic acids, and glycans is critical for biopharmaceutical research and quality control. Complex samples require multiple chromatographic techniques to fully elucidate structure and heterogeneity.
The ACQUITY UPLC H-Class Bio System extends ultrahigh-performance liquid chromatography (UPLC) to biomolecular analysis, integrating multi-mode separation (reversed phase, ion exchange, size exclusion, HILIC) on a single platform. This summary highlights system design, workflow automation, and chromatographic performance.
Key system components include:
Automated buffer blending using Auto•Blend Plus enabled rapid optimization of pH and ionic strength for ion exchange separations, demonstrated by separation of a three-protein mixture with clear peak resolution across pH gradients. UPLC peptide mapping showed superior resolution and sensitivity compared to HPLC. Size exclusion analysis of aggregates versus monomeric IgG was completed in under 3.7 minutes with enhanced peak sharpness. HILIC separation of 2-aminobenzamide-labeled glycans from human IgG achieved baseline resolution of major glycoforms.
• High resolution and sensitivity from sub-2 µm columns with minimized system dispersion improve detection of low-abundance variants.
• Increased throughput reduces analysis time and sample reanalysis, lowering costs and accelerating decision-making.
• Robust operation under high ionic strength and varied solvent conditions ensures reproducibility.
• Flexible multi-mode capability on a single system simplifies workflows for peptide mapping, intact protein characterization, glycan profiling, and oligonucleotide analysis.
Upcoming developments include tighter integration with mass spectrometry workflows, enhanced informatics and AI-driven method development, novel column chemistries for expanded biomolecule classes, and increased automation for high-throughput biopharma screening and QC environments.
The ACQUITY UPLC H-Class Bio System delivers a comprehensive, flexible, and automated solution for high-performance biomolecular separations, combining UPLC speed and resolution with robust multi-mode capability to support advanced biopharmaceutical analysis.
HPLC
IndustriesManufacturerWaters
Summary
Significance of the topic
Accurate and rapid characterization of biological macromolecules such as proteins, peptides, nucleic acids, and glycans is critical for biopharmaceutical research and quality control. Complex samples require multiple chromatographic techniques to fully elucidate structure and heterogeneity.
Aims and overview
The ACQUITY UPLC H-Class Bio System extends ultrahigh-performance liquid chromatography (UPLC) to biomolecular analysis, integrating multi-mode separation (reversed phase, ion exchange, size exclusion, HILIC) on a single platform. This summary highlights system design, workflow automation, and chromatographic performance.
Methodology and instrumentation
Key system components include:
- Quaternary Solvent Manager with Auto•Blend Plus technology for on-demand blending of pure solvents and concentrated buffer stocks to achieve precise pH and ionic strength gradients.
- Flow-through-needle Sample Manager with inert flow path for high recovery and minimal carryover.
- Column heaters and multi-column managers ensuring accurate temperature control and low dispersion.
- UPLC-optimized detectors: photodiode array, UV/Vis, fluorescence, evaporative light scattering, quadrupole and time-of-flight mass spectrometers.
- Empower™ and MassLynx™ software with BiopharmaLynx™ for data acquisition and analysis.
Main results and discussion
Automated buffer blending using Auto•Blend Plus enabled rapid optimization of pH and ionic strength for ion exchange separations, demonstrated by separation of a three-protein mixture with clear peak resolution across pH gradients. UPLC peptide mapping showed superior resolution and sensitivity compared to HPLC. Size exclusion analysis of aggregates versus monomeric IgG was completed in under 3.7 minutes with enhanced peak sharpness. HILIC separation of 2-aminobenzamide-labeled glycans from human IgG achieved baseline resolution of major glycoforms.
Benefits and practical applications
• High resolution and sensitivity from sub-2 µm columns with minimized system dispersion improve detection of low-abundance variants.
• Increased throughput reduces analysis time and sample reanalysis, lowering costs and accelerating decision-making.
• Robust operation under high ionic strength and varied solvent conditions ensures reproducibility.
• Flexible multi-mode capability on a single system simplifies workflows for peptide mapping, intact protein characterization, glycan profiling, and oligonucleotide analysis.
Future trends and potential applications
Upcoming developments include tighter integration with mass spectrometry workflows, enhanced informatics and AI-driven method development, novel column chemistries for expanded biomolecule classes, and increased automation for high-throughput biopharma screening and QC environments.
Conclusion
The ACQUITY UPLC H-Class Bio System delivers a comprehensive, flexible, and automated solution for high-performance biomolecular separations, combining UPLC speed and resolution with robust multi-mode capability to support advanced biopharmaceutical analysis.
Used instrumentation
- ACQUITY UPLC H-Class Bio System with quaternary solvent manager and flow-through-needle injector
- Auto•Blend Plus buffer blending
- Column heaters and multi-column managers
- UPLC-optimized detectors (PDA, UV/Vis, fluorescence, ELS, MS)
- Empower™ and MassLynx™ with BiopharmaLynx™
Content was automatically generated from an orignal PDF document using AI and may contain inaccuracies.
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