(T1230-06-36) Quantitative Assessment of mRNA Drug Biological Activity by Cell-Based Assays

Purpose: Biological activity of mRNA drugs, historically assessed via animal study, is critical to drug development, mechanistic evaluation of drug candidates, and as a manufacturing critical quality attribute (CQA). However, animal testing is costly, time-consuming and presents a significant environmental impact highlighted by the recent passing of Bill S.5002 - FDA Modernization Act 2.0 which aims to mediate the ecological impact of drug discovery and development by reducing animal use. The aim of this study is to develop quantitative assessment of mRNA drug biological activity by cell-based sssays.

Methods: In response to the need for robust in-vitro assays which can serve as a proxy for activity of mRNA therapeutics, we report successful development of two in-cell Western assays that assess the biological activity of a bivalent mRNA vaccine against Sars-CoV-19.  We used the A549 cell line to evaluate the antigenicity of specific binding antibodies. We standardized the experimental procedure and analyzed the data with 4PL models.

Results: These assays meet the ICH M10 guideline for large molecule PK study with coefficient of variation (CV < 20%) and relative error (RE < 20%) for both accuracy and precision.

Conclusion: These sensitive, reproducible assays hold great potential not only to GMP testing but also to large scale screening of mRNA drugs.

Acknowledgements: No conflict declared.