Host cell protein quantification workflow using optimized standards combined with data-independent acquisition mass...

Area of Science:

• Biopharmaceutical manufacturing
• Analytical chemistry
• Protein analysis

Background:

• Host cell protein (HCP) monitoring is vital for monoclonal antibody (mAb) safety and efficacy.
• Traditional ELISA methods for HCP quantification have limitations in protein identification.
• Mass spectrometry (MS) offers a powerful alternative for qualitative and quantitative HCP analysis.

Purpose of the Study:

• To develop and validate a sensitive and robust MS-based workflow for HCP quantification in mAb production.
• To evaluate the performance of the HCP Profiler solution and a data-independent acquisition (DIA) method.
• To establish a standardized analytical method for biopharmaceutical process development and quality control.

Main Methods:

• Implementation of a liquid chromatography-MS (LC-MS) based workflow.
• Utilizing an innovative quantification standard, the HCP Profiler solution.
• Employing a spectral library-based data-independent acquisition (DIA) method with strict data validation.

Main Results:

• The MS-based workflow demonstrated high accuracy and reproducibility (CV < 10%) for HCP quantification.
• Achieved high sensitivity, detecting HCPs down to the sub-ng/mg mAb level.
• The spectral library-based DIA approach outperformed data-dependent acquisition and library-free DIA interpretation.

Conclusions:

• The presented MS-based workflow is a mature and robust method for HCP monitoring in mAb manufacturing.
• This approach supports process development and ensures drug product quality control.
• The HCP Profiler solution and DIA method offer significant advantages over conventional techniques.