Sensitive, Rapid, Robust, and Reproducible Workflow for Host Cell Protein Profiling in Biopharmaceutical Process...

Area of Science:

• Biopharmaceutical Manufacturing
• Analytical Chemistry
• Proteomics

Background:

• Host cell proteins (HCPs) are critical impurities in biopharmaceutical production.
• Regulatory agencies require HCP identification and quantification for risk assessment.
• Existing HCP detection methods can be time-consuming, limiting process development integration.

Purpose of the Study:

• To develop a sensitive, robust, and reproducible workflow for HCP detection and quantification.
• To significantly reduce the turnaround time for HCP analysis.
• To enable high-throughput HCP profiling for biopharmaceutical process development.

Main Methods:

• Utilized an Evosep ONE LC system coupled to an Orbitrap Fusion Lumos mass spectrometer.
• Developed a rapid workflow for HCP detection and quantification.
• Analyzed NIST monoclonal antibody reference material.

Main Results:

• Achieved significantly shorter turnaround times compared to previous methods.
• Enabled high-throughput analysis with a capacity of 60 samples per day.
• Detected the largest number of HCPs reported to date in the NIST reference material, demonstrating improved performance and throughput.

Conclusions:

• The developed HCP workflow is sensitive, robust, and reproducible.
• The rapid nature of the workflow allows for integration into biopharmaceutical process development.
• This method facilitates better risk assessment and optimization of biopharmaceutical production by enabling rapid measurement and clearance tracking of HCPs.