BEBPA

Orthogonal HCP ELISA Characterization and Supply
Control Strategy to Support Marketed Drug Product

Use of a generic off-the-shelf HCP ELISA has several challenges both before and after it is validated for use in bioprocess monitoring. First, it is difficult demonstrate that an HCP ELISA is fit-for-purpose to use in bioprocess control strategy and second consistent long-term reagent supply is an established risk for HCP ELISAs.

To validate HCP reagents, ELISA and 2D coverage are common orthogonal techniques that together demonstrate that the anti-HCP ELISA is fit for purpose. Mass spectrometry provides another way to assess HCP impurities, and unlike ELISA provides identification of the individual HCP proteins. As such mass spectrometry is now commonly used as both a primary and an orthogonal method for HCP assessment along with 2D “DIGE/DIBE immunoblots and ELISA.

Here we describe the characterization of a generic CHO HCP assay using standard methods of 2D, ELISA, coupled with an orthogonal LC-MS analysis of the HCP standard and the evaluation of potential problematic HCPs as part of the qualification. We also propose recommendations for demonstrating fit-for-purpose to the specific bioprocess and for securing long term HCP reagent supply to reduce the risk of premature critical reagent depletion.

Presentation given by: Rob Hooper, Senior Research Scientist
(Day 1: Implementation of HCP ELISAs: Commercial, Platform, & Process-Specific Assays Session)