A high density assay format for the detection of novel cytotoxic  agents in large chemical libraries.
In response to the need for inexpensive high throughput assays for anti-cancer drug screening, a 1536-well microtiter plate based assay utilizing the Alamar Blue fluorescent dye as a measure of cellular growth was validated in 10 microL assay volume. Its robustness was assessed in a screen against a library of 2000 known bioactives; with an overall Z' value of 0.89 for assay robustness, several known cytotoxic agents were identified including and not limited to anthracyclines, cardiac glycosides, gamboges, and quinones. To further test the sensitivity of the assay, IC50 determinations were performed in both 384-well and 1536-well formats and the obtained results show a very good correlation between the two density formats. These findings demonstrate that this newly developed assay is simple to set up, robust, highly sensitive and inexpensive. It could potentially provide a rapid way to screen established and primary tumor cell lines against large chemical libraries.
Shum
David
D
High Throughput Screening Core Facility, Memorial Sloan-Kettering Cancer Center, 1275 York Avenue, New York, NY 10021, USA.
Radu
Constantin
C
Kim
Earl
E
Cajuste
Muriel
M
Shao
Yufang
Y
Seshan
Venkatraman E
VE
Djaballah
Hakim
H
D002460:Cell Line; D049109:Cell Proliferation / Q000187:drug effects; D020128:Inhibitory Concentration 50; D015394:Molecular Structure; D054852:Small Molecule Libraries / Q000032:analysis* / Q000737:chemistry* / Q000633:toxicity
D054852:Small Molecule Libraries