Comparison of Three Luciferase-Based Detection Kits for Cell Viability Assessment in a Cell-Based Potency Assay

Purpose

Potency assays play a crucial role in the development of biologic drugs, ensuring the strength and effectiveness of a drug product. The accuracy of potency assays is fundamental in determining the appropriate dosage and in ensuring the safety and efficacy of the drug, both in preclinical studies and clinical trials.

Cell viability assays, commonly used to assess potential cytotoxicity or therapeutic effects of a drug, measure the health, survival, and metabolic activity of cells in the presence of a drug. These assays provide insights into how a drug affects cellular function and survival. The success and accuracy of cell viability assays depend on the critical reagents used to assess cell health. These reagents are essential in evaluating the potency—the key quality attribute of biologics. To ensure reliable results, the performance of viability detection reagents must be continuously monitored throughout the potency assay’s lifecycle, accounting for potential sources of variance and their impact on assay outcomes over time.

Cell viability assays use a variety of markers as indicators of metabolically active (living) cells. Examples of markers commonly used include measuring ATP levels,  measuring the ability to reduce a substrate, and detecting enzymatic/protease activities unique to living cells. Firefly luciferase, which measures ATP concentrations for cell viability, is widely used due to its easy use, speed, higher sensitivity, and less prone to artifacts than other methods.

The current study compared cell viability detection using firefly luciferase-based assays from three different vendors with a qualified cell-based proliferation assay.