ELISA reagent characterization using advanced LC-MS methods

When using commercial or process-specific ELISAs for Host Cell Protein (HCP) impurity measurements, regulatory reviewers require a characterization of the suitability of the ELISA reagents to the specific product.

Our observation from running a vast number of projects is that frequent challenges in ELISAs are:

A) Low HCP coverage
B) ELISA not measuring abundant HCPs
C) lack of dilutional linearity
D) different HCP values obtained when using new ELISA reagents

2D PAGE analysis cannot answer these challenges; they require higher resolution analytical methods to provide detailed information about the HCP coverage.

This webinar will show you results from ELISA reagent characterization using advanced LC-MS techniques on various biologics and expression systems.
​​​​​
​​​​​​The analyses include:

• HCP Coverage analysis of early process sample using ELISA immunocapture combined with mass spectrometry (ELISA-MS)
• Coverage of individual HCPs of potential concern
• Coverage of individual HCPs in the purified drug substance
• Comparison of the HCP standard with the early process sample
• Evaluation of high-responding HCPs and non-specific binding likely leading to lack of dilutional linearity
• Evaluation of cross-reactivity to the drug substance

Such a comprehensive ELISA reagent characterization enables a detailed understanding of the HCP-ELISA reagents and documentation of their suitability to the specific manufacturing process and purified drug substance. The LC-MS analyses also help bridge between ELISA reagents and explain differences in HCP measurements between old and new reagents.