Intelligent Data Capture (IDC) Enables Optimal Xevo G2-XS Data Acquisition and Processing for Multi-Attribute Method (MAM) Studies

Importance of the Topic

Multi-Attribute Method (MAM) is critical for direct measurement of product and critical quality attributes in biopharmaceutical development, enabling simultaneous attribute monitoring, impurity detection, and new peak identification.

Objectives and Study Overview

This study evaluates the impact of the Intelligent Data Capture (IDC) noise reduction algorithm on the Xevo G2-XS QTof system for peptide-level MAM analysis. Key aims include assessing quantitative fidelity, new peak detection performance, raw data file size reduction, and data processing speed across IDC threshold settings (Off, 5, 10, 15).

Methodology

Sample Set:

• System suitability: MassPREP peptide mixture
• Control: Tryptic digestion mAb standard
• Spiked control: mAb digest with 15 heavy-labeled peptides
• Stressed sample: NISTmAb incubated at pH 8.0, 40 °C for 14 days then digested

LC Conditions:

• ACQUITY UPLC H-Class PLUS Bio System
• ACQUITY Premier Peptide CSH C18 column, 60 °C, 0.2 mL/min
• Mobile phases: 0.1% formic acid in water (A), 0.1% formic acid in acetonitrile (B)
• Injection volume: 10 µL; sample temperature: 6 °C

MS Conditions:

• Xevo G2-XS QTof, ESI+ in MSE mode, m/z 50–2000
• Capillary voltage: 1.2 kV; cone voltage: 20 V; collision energy ramp: 60–120 V

Data Management:

• waters_connect platform with Peptide MAM and UNIFI applications

Used Instrumentation

• ACQUITY UPLC H-Class PLUS Bio System
• ACQUITY UPLC Tunable UV Detector
• Xevo G2-XS QTof MS
• waters_connect informatics with Peptide MAM and UNIFI apps

Main Results and Discussion

Quantitative Fidelity:

• % modification profiles for 30 attributes, including low-abundance glycopeptides (Man5 at ~0.08%), remained consistent across IDC Off and IDC15 settings.
• IDC15 spectra demonstrated reduced baseline noise and preserved isotope distributions for weak signals.

New Peak Detection:

• IDC15 acquisition identified all true new peaks in spiked and stressed samples while eliminating false positives observed at IDC5 and IDC Off.
• Lower IDC thresholds and no IDC increased manual validation workload due to artifacts.

Data Reduction and Processing Efficiency:

• IDC5 reduced raw file size by 65%; IDC15 achieved over 90% reduction versus IDC Off.
• Data processing times decreased by up to 75% at IDC15, saving more than three hours for two injections.

Benefits and Practical Applications

• Streamlined, compliance-ready peptide MAM workflows with minimal manual intervention.
• Enhanced confidence in new peak detection, reducing regulatory risks and out-of-specification investigations.
• Significant data storage savings and faster data analysis without loss of sensitivity or quantitation accuracy.

Future Trends and Applications

• Extension of real-time noise reduction algorithms to other mass spectrometry platforms and omics workflows.
• Integration of AI-driven data analysis for adaptive thresholding and enhanced feature extraction.
• Wider regulatory acceptance of MAM workflows embedded with intelligent data reduction and automated reporting.

Conclusion

Implementing IDC on the Xevo G2-XS QTof delivers reproducible peptide MAM results while dramatically reducing raw data sizes and processing times. The default IDC15 setting offers optimal noise suppression, eliminates false-positive new peaks, and enhances overall workflow efficiency, supporting robust quality monitoring in biopharmaceutical development.

Reference

1. Mortishire-Smith R, Richardson K, Denny R, Hughes C. Intelligent Data Capture: Real-Time Noise Reduction for High Resolution Mass Spectrometry. Waters White Paper, 720006567EN (2019).
2. Ippoliti S, Yu YQ, Mortishire-Smith R. Peptide Mapping Using Intelligent Data Capture on Vion IMS QTof. Waters Application Brief, 720006636EN (2019).