Host cell protein detection gap risk mitigation: quantitative IAC-MS for ELISA antibody reagent coverage...

Area of Science:

• Biopharmaceutical Manufacturing
• Proteomics
• Analytical Chemistry

Background:

• Host cell proteins (HCPs) in biopharmaceuticals require strict clearance for patient safety.
• Enzyme-linked immunosorbent assays (ELISAs) using polyclonal antibodies are standard for monitoring HCP clearance.
• Assessing the broad coverage of these polyclonal antibodies is crucial for ELISA reliability.

Purpose of the Study:

• To introduce and validate a quantitative proteomics approach combined with immunoaffinity chromatography (qIAC-MS) for assessing ELISA antibody coverage.
• To investigate if HCPs used for animal immunization represent process-relevant HCPs.
• To compare qIAC-MS with traditional gel-based methods for evaluating ELISA coverage.

Main Methods:

• Characterization of cell culture fluid (CCF) from mock fermentation and a recombinant monoclonal antibody product.
• Application of quantitative immunoaffinity chromatography coupled with mass spectrometry (qIAC-MS).
• Utilized controls to identify HCPs in reagents and non-specific binding.

Main Results:

• The qIAC-MS approach was demonstrated as a suitable orthogonal method to 2D SDS-PAGE for evaluating ELISA antibody coverage.
• Identified specific HCPs covered and missed by the existing ELISA.
• Quantified HCPs present in the polyclonal reagent and those binding non-specifically.

Conclusions:

• The qIAC-MS method provides a more comprehensive assessment of ELISA antibody coverage than traditional methods.
• This technique enables a more refined risk assessment of HCPs in biopharmaceutical products.
• Improved understanding of HCP coverage enhances overall product quality and patient safety.