Development and Comparative Analysis of a 384-well Luciferase-Based Viability HTS Assay for Trypanosoma brucei brucei BS427 cells

摘要

We describe the development of a whole cell, high throughput (HTS) assay for Trypanosoma brucei brucei whole cells. This development contributes to the early drug discovery process for Human African Trypanosomiasis (HAT). The assay is in 384 well format and estimates cell viability via a luciferase based reagent to detect intracellular ATP. The assay fulfils statistical requirements for reproducibility with a Z' of >0.5. Criteria that were explored to define parameters for a reproducible assay included cell inoculum, final cell density, DMSO sensitivity and compound diluent. A panel of reference compounds were found to have comparable activity to published assay protocols and in house Alamar Blue assay in 384 well format.