Performing the biopharmaceutical multi-attribute method (MAM)

Importance of Topic

The multi attribute method MAM represents a significant advancement in biopharmaceutical analysis. It combines high resolution accurate mass LC MS with targeted quantitation and new peak detection in a single workflow. This capability addresses the growing demand for robust characterization and monitoring of critical quality attributes in therapeutic proteins across development and manufacturing.

Objectives and Overview

This technical note describes a comprehensive HR MAM workflow integrating BioPharma Finder and Chromeleon Chromatography Data System. It outlines the transition from discovery phase attribute characterization to compliance ready CQA monitoring and new peak detection. The focus is on data acquisition, processing, and reporting strategies that ensure consistent results from research to quality control laboratories.

Methodology

• Peptide mapping in BioPharma Finder to achieve full sequence coverage and identify product quality attributes including post translational modifications.
• Selection of critical quality attributes and creation of a target peptide workbook for targeted MS1 quantitation.
• Import of the peptide workbook into Chromeleon CDS and optimization of MS detection settings retention time alignment and isotope integration parameters.
• Relative quantitation of CQAs based on integrated peak areas within predefined report templates.
• Non targeted full scan MS processing for new peak detection using frame alignment filtering and reporting functions.

Instrumentation

• Thermo Scientific Vanquish Horizon UHPLC system
• Thermo Scientific Q Exactive Plus Hybrid Quadrupole Orbitrap or Exactive Plus Orbitrap mass spectrometer
• Thermo Scientific BioPharma Finder software
• Thermo Scientific Chromeleon Chromatography Data System

Key Results and Discussion

The MAM workflow demonstrated robust identification of product quality attributes with high resolution MS MS and reliable mapping of sequence coverage. The target peptide workbook facilitated consistent monitoring of selected CQAs including glycoforms and oxidation variants. Optimization of detection parameters in Chromeleon CDS enabled accurate integration of low abundance modifications such as deamidation. New peak detection efficiently highlighted unexpected impurities by comparing test samples against reference standards and applying filtering rules to minimize false positives.

Benefits and Practical Applications

The integrated software approach supports seamless method transfer from research to regulated environments. Compliance ready acquisition and data audit trails in Chromeleon CDS ensure full traceability. Customizable report templates and automated workflows streamline routine quality control testing of biotherapeutics while maintaining high data integrity and reproducibility.

Future Trends and Possibilities

Advancements in data processing algorithms and machine learning are expected to enhance automated impurity detection and attribute quantitation. Integration with laboratory information management systems will further accelerate sample tracking and result review. Expansion of MAM techniques to emerging modalities such as cell and gene therapies will drive broader adoption across the biopharma industry.

Conclusion

The HR MAM methodology leveraging BioPharma Finder and Chromeleon CDS delivers a powerful platform for comprehensive characterization and monitoring of biotherapeutic quality attributes. By combining targeted quantitation with new peak detection in a compliant environment the workflow ensures consistent data generation from early development through commercial quality control.

Reference

1. Rogers R et al Development of a quantitative mass spectrometry multi attribute method for characterization quality control testing and disposition of biologics Mabs 2015 7 881
2. Rogers R et al A view on the importance of multi attribute method for measuring purity of biopharmaceuticals and improving overall control strategy The AAPS Journal 2018 20 7
3. Rogstad S et al Multi attribute method for quality control of therapeutic proteins Anal Chem 2019 91 14170
4. Liu HC et al A high resolution accurate mass multi attribute method for critical quality attribute monitoring and new peak detection Thermo Scientific Application Note 72916