ZipChip™ System for Selected Thermo Scientific™ Mass Spectrometers - for CE/ESI-MS Analyses

Importance of the Topic

Capillary electrophoresis electrospray ionization mass spectrometry CE ESI MS is a powerful approach for rapid high resolution separation of charged and polar analytes in biopharmaceutical quality control metabolomics and biomarker discovery applications

Objectives and Overview of the Study

This application note introduces the ZipChip system for integrated microfluidic CE separation and on chip ESI into Thermo Scientific mass spectrometers The note demonstrates workflows covering intact antibody profiling ADC analysis subunit mapping peptide analysis and metabolite quantification

Methodology and Instrumentation

• ZipChip interface and microfluidic chip assembly allowing CE separation and electrospray on chip
• ZipChip autosampler for automated sample loading priming and flushing
• DryDock accessory for chip drying and storage
• ZipChip HR and HS chips with 22 cm and 10 cm channels on board desalting emitter and usage tracking
• Assay kits for intact antibodies peptides and metabolites providing buffers and diluents
• Mass spectrometers Q Exactive Plus HF HF X with BioPharma Option focusing on HRAM detection in intact and denatured states

Key Results and Discussion

• Intact mAbs trastuzumab infliximab NIST mAb achieved sub 3 minute separation with baseline resolution of charge variants lysine clipping and major glycoforms at pg to ng consumption
• ADC analysis of trastuzumab emtansine resolved drug to antibody ratio DAR0 to DAR8 in under one minute with average DAR consistent with literature
• Subunit profiling via IdeS digest and reduction allowed mapping Fc2 Fd and light chain subunits in 3 minutes identifying glycoforms and clipping variants
• Peptide mapping delivered 98 percent sequence coverage for heavy and light chains in 10 minutes with a few nanograms of digest
• Metabolomics assays separated 20 amino acids including isomers leucine isoleucine in 2 minutes with linear dynamic range from 10 nM to 10 µM and low carryover
• Direct analysis of amino acids in high salt cell culture media achieved via on chip desalting enabling rapid quantification with stable isotope internal standard

Benefits and Practical Applications

• Extremely fast high resolution separations combined with nano spray sensitivity and HRAM detection
• Minimal sample preparation on chip desalting supports high salt samples
• Low sample consumption down to femtograms improves throughput and cost efficiency
• Broad application covering biotherapeutics metabolomics peptide analysis QA QC and industrial analytics

Future Trends and Potential Applications

The compact integrated CE ESI MS platform can evolve with higher throughput chip formats expanded assay kits and advanced software for real time data processing enabling label free quantification deeper glycoanalytics and on line quality control in biomanufacturing

Conclusion

The ZipChip system offers a portable complete CE ESI solution for direct coupling to mass spectrometers delivering rapid robust reproducible separations and high quality MS data across diverse application areas in analytical chemistry and biopharma quality control