Monitoring Host cell proteins during bioprocess development and biologics manufacturing is a critical task often performed using ELISA. Custom process-specific HCP ELISA are typically the preferred method for measuring HCPs. However, it is not always appropriate or practical to generatede novoantibodiesand a customized HCP ELISA. This creates a strong need for robust, ready-to-use, generic HCP assays to support biologics development.
Here we discuss the generation and validation of a generic CHO-specific HCP immunoassay. We describe the overall workflow starting from creation of the representative HCCF as the HCP reference standard, the development of a broad coverage CHO-HCP antibody, use of multiple orthogonal methods (including amino acid analysis, 2D Western blot and 2D DIBE coverage assay, as well as mass spec) for characterization of the HCP coverage. Lastly, we describe validation of the ELISA for in-process samples and final drug substance as well as supply chain management to ensure long-term access to the validated assay components.