Cell-Based Assay (CBA)

Using PIPETMAX® 268 to Maximize Informational Content from Cell-Based Assays using Multi-Parameter Mechanistic Toxicity Profiling.

Abstract/Summary

Cell-Based Assay: Testing compounds of interest in the biological context of the cell is a necessary step to identify toxic liability or confirm mechanism of action. In this application, a mechanistic toxicity profiling panel has been created in 384-well format consisting of assays used together to evaluate cell viability, cytotoxicity, and apoptosis. The assays consist of detection chemistries for live and dead cell protease activity, ATP content, membrane integrity, and caspase-3/7 activity.

In order to prevent manual errors from impacting Z’-factor results, PIPETMAX 268, an automated pipetting station, was used to perform all steps of experimental set-up including cell dispensing, serial titration and array of compound, and addition of the assay reagents to the plate. This shows representative data with K562 (human, BCR-ABL+ CML model) and HepG2 (human, hepatocellular carcinoma) cell lines, demonstrating dose and time-dependent toxicity profiles for anti-proliferative, necrosis and apoptosis-inducing compounds and discussed the Z’-factors achieved in each assay.

Legend: A five-assay profile was used to distinguish necrosis, apoptosis, and cytostasis in K562 Cells

Legend: A five-assay profile was used to distinguish necrosis, apoptosis, and cytostasis in K562 Cells

 


Cell Based Assay (CBA)
The CBA application poster that resulted from the collaboration between Gilson, Inc and Promega Corp.


To See the CBA Data:

Download Application Poster