A Cell-Based Potency Assay for Determining the Relative Potency of Botulinum Neurotoxin A Preparations Using Manual and Semi-Automated Procedures

Cell-based potency assays (CBPAs) are required for the potency testing and commercial release of botulinum neurotoxin (BoNT)-based drug products. These CBPAs must account for the toxin's biological activities while meeting regulatory guidelines for precision and accuracy. Here, studies describe the characterization and qualification of the BoSapient CBPA and demonstrate that it is fit for use as a relative potency assay for BoNT/A-containing samples. The CBPA is operated in a 96-well plate format and relies upon the fluorescence emissions of a reporter that directly responds to BoNT/A activity. The BoSapient cell line expresses the BoNT/A-receptors SV2 and complex gangliosides, is responsive only to intact BoNT/A, and can robustly detect picomolar and sub-picomolar BoNT/A quantities, making the CBPA appropriate for quantifying BoNT/A-based drug products. The cell line was passaged 30 times without significant loss of reporter expression or BoNT/A sensitivity. Manual and semi-automated CBPA methods were developed and qualified according to regulatory guidelines and shown to have low bias (