Those cells have been used to develop a bioanalytical system which we are currently characterizing. This report details recent progress in adapting the RLT 2.0 bioassay to a 96-well microplate, and optimizing the assay for high throughput and efficiency. It also provides RLT 2.0-derived potency estimates for several dioxin and furan congeners, not tested previously, and describes the results of a sensitivity analysis used to estimate the range of uncertainty associated with application of those estimates. The RLT 2.0 bioassay was successfully adapted to and optimized for a 96-well microplate format.
Analysis is performed using a microplate reading luminometer.
In vitro protein assay performed after luciferase assay, provides a secondary quantitative measure of potential differences in cell number and thus viability per well. Overall, the 96-well RLT 2.0 bioassay developed greatly increased assay efficiency and throughput without significant loss of sensitivity. RLT 2.0-derived relative potencies were similar to those based on other rainbow trout specific and mammalian bioassays. Thus, a sensitivity analysis was conducted to estimate the range of uncertainty associated with TEQ estimates based on RLT 2.0-derived relative potencies. Richter, C.A., V.L. Tieber, M.S. Denison, and J.P. Giesy.