Chemiluminometric biochemical induction assay (CBIA) for the detection of DNA-damaging agents.
A microbroth chemiluminometric version of the biochemical induction assay (BIA) was developed using a chemiluminescent substrate widely used to detect beta-galactosidase in high-throughput screening (HTS) laboratories. The assay was run in both 96-well and 384-well plate formats using the Zymark RapidPlate liquid handling system to transfer samples and reagents. Chemiluminescence was read using the Victor-2 multilabel counter. The new microbroth chemiluminometric method, the CBIA, allowed rapid screening of samples, crude extracts, and pure compounds for their DNA-damaging effects in bacteria. In screening a small subset of our natural products library samples by the agar plate BIA and the CBIA, the latter yielded a higher hit rate, suggesting it is more sensitive than the agar plate assay. The CBIA was unaffected by the colored samples often encountered during screening of crude natural products extracts.
Singh
Maya Prakash
MP
Natural Products, Chemical and Screening Sciences, Wyeth Research, Pearl River, NY 10965, United States. singhm@wyeth.com
Arias
Daniel A
DA
Greenstein
Michael
M
D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D004249:DNA Damage; D004269:DNA, Bacterial / Q000187:drug effects*; D004926:Escherichia coli / Q000187:drug effects* / Q000235:genetics / Q000378:metabolism; D008163:Luminescent Measurements / Q000379:methods*; D001616:beta-Galactosidase / Q000032:analysis
D000900:Anti-Bacterial Agents; D004269:DNA, Bacterial; D001616:beta-Galactosidase