Enabling Intelligent Data Capture for Intact Mass Analysis of Monoclonal Antibodies and Subunits using a Vion IMS QTof HRMS

Importance of the Topic

In intact monoclonal antibody and subunit analysis high resolution mass spectrometry generates rich data sets that require efficient handling. Intelligent Data Capture reduces noise and background in real time, lowering file sizes while preserving spectral quality.

Objectives and Study Overview

This study evaluates the effect of enabling Intelligent Data Capture on data quality metrics and file size for intact trastuzumab and NIST mAb subunits across varying IDC thresholds (5, 10, and 20 counts) compared to no IDC.

Methodology

• Samples: Trastuzumab at 100 micrograms per milliliter in ammonium acetate and BSA solution; NIST mAb subunit standard at 100 micrograms per milliliter in water.
• Acquisition: Electrospray ionization time of flight MS in positive full scan mode with IDC disabled and enabled at thresholds of 5, 10, and 20 counts.
• Deconvolution: MaxEnt1 software applied to combined MS spectra for mass assignment and glycoform distribution analysis.

Used Instrumentation

• Waters Vion IMS QTof High Resolution Mass Spectrometer with Intelligent Data Capture option.
• ESI-TOF ion source configured for positive ion full scan.

Main Results and Discussion

Enabling IDC at the default threshold of 5 counts yields a 39 percent reduction in file size for intact trastuzumab, with further reductions to 44 and 59 percent at thresholds of 10 and 20 counts, respectively. Glycoform relative abundances and charge state distributions remain consistent across modes. For reduced mAb subunits, IDC provides 45 percent file size reduction at 5 counts and up to 80 percent at 20 counts, with no compromise in spectral integrity.

Practical Benefits and Applications

• Substantial reduction in data storage requirements and transfer times.
• Improved processing throughput for routine LC MS workflows.
• Maintained confidence in qualitative and quantitative outcomes.

Future Trends and Potential Applications

Integration of adaptive threshold algorithms, expansion to broader proteomic workflows, and leveraging cloud based data management may further enhance the utility of real time noise reduction in high throughput laboratories.

Conclusion

Intelligent Data Capture delivers an efficient solution for reducing file sizes by up to 80 percent while preserving the critical spectral features required for intact antibody and subunit mass analysis. The default threshold of five counts provides an optimal balance between noise reduction and data fidelity.

References

1. Intelligent Data Capture Real Time Noise Reduction for High Resolution Mass Spectrometry Waters white paper 720006567EN
2. High Performance Vion IMS QTof for Monoclonal Antibody Intact and Subunit Mass Analysis Waters application note 720005906EN
3. High Sensitivity Intact Monoclonal Antibody HRMS Quantification Waters application note 720006222EN