Quantitative host cell protein analysis using two dimensional data independent LC-MS(E)
Amy Farrell1,2, Stefan Mittermayr1, Brian Morrissey1
1Characterisation and Comparability Laboratory, NIBRT-The National Institute for Bioprocessing Research and Training , Foster Avenue, Mount Merrion, Blackrock, Co., Dublin, Ireland.
Analytical Chemistry
|August 18, 2015
Summary
Host cell proteins (HCPs) are impurities in biotherapeutics. Mass spectrometry effectively identifies and quantifies HCPs, guiding purification strategies to minimize patient risk and ensure therapeutic protein safety.
Area of Science:
- Biopharmaceutical Manufacturing
- Proteomics
- Analytical Chemistry
Background:
- Host cell proteins (HCPs) are critical impurities in recombinant protein production, necessitating robust removal strategies.
- Traditional anti-HCP ELISA methods offer limited characterization, prompting the need for advanced analytical techniques.
- Ensuring patient safety requires thorough assessment and control of HCPs in therapeutic proteins.
Purpose of the Study:
- To characterize Host cell protein (HCP) repertoires in monoclonal antibody (mAb) samples using advanced mass spectrometry.
- To evaluate the impact of downstream purification (DSP) parameters, including harvest time and elution buffers, on HCP levels.
- To assess the effect of purification strategies on critical quality attributes (CQAs) of therapeutic mAbs.
Main Methods:
- Application of a 2D-LC-MS(E) proteomic platform for high-resolution HCP profiling.
- Analysis of HCPs in Protein A purified mAb samples across different harvest times and elution buffers.
- Evaluation of CQAs for mAbs purified using various Protein A elution buffers.
Main Results:
- An arginine-based Protein A elution buffer significantly minimized identified and quantified HCP levels in purified mAb samples.
- Later harvest times correlated with higher HCP concentrations, which were effectively reduced by complementary DSP steps.
- Purification strategies employing specific elution buffers and DSP steps did not adversely affect mAb product CQAs.
Conclusions:
- Mass spectrometry-based HCP determination provides valuable insights for optimizing purification processes.
- Strategic selection of Protein A elution buffers and downstream purification steps is crucial for minimizing HCPs in therapeutic mAbs.
- Optimized bioprocessing parameters, guided by mass spectrometry, enhance the safety and efficacy of therapeutic proteins by reducing HCP burden.
