Feasibility of a High-Flux Anticancer Drug Screen Using a Diverse Panel of Cultured Human Tumor Cell Lines

Abstract

We describe here the development and implementation of a pilot-scale, in vitro, anticancer drug screen utilizing a panel of 60 human tumor cell lines organized into subpanels representing leukemia, melanoma, and cancers of the lung, colon, kidney, ovary, and central nervous system. The ultimate goal of this disease-oriented screen is to facilitate the discovery of new compounds with potential cell line-specific and/or subpanel-specific antitumor activity. In the current screening protocol, each cell line is inoculated onto microtiter plates, then preincubated for 24-28 hours. Subsequently, test agents are added in five 10-fold dilutions and the culture is incubated for an additional 48 hours. For each test agent, a dose-response profile is generated. End-point determinations of the cell viability or cell growth are performed by in situ fixation of cells, followed by staining with a protein-binding dye, sulforhodamine B (SRB). The SRB binds to the basic amino acids of cellular macromolecules; the solubilized stain is measured spectrophotometrically to determine relative cell growth or viability in treated and untreated cells. Following the pilot screening studies, a screening rate of 400 compounds per week has been consistently achieved.

Controlling complement resistance in cancer by using human monoclonal antibodies that neutralize complement-regulatory proteins CD55 and CD59

“…Briefly, 2Â10 5 /50 ll cells were incubated with Rituximab at 2 lg/ml with or without blocking antibodies (10 lg/ml) to a final volume of 100 ll for 10 min at room temperature prior to the addition of NHS (25%). Residual viable cells were measured after 1 h at 37 C using an MTT assay [38].…”

Involvement of c-Fos in fenretinide-induced apoptosis in human ovarian carcinoma cells

“…Cell number was estimated by using the sulforhodamine B assay. 48 Three analyses were performed, and four replicate wells were used for each analysis.…”

Mining and Visualizing Large Anticancer Drug Discovery Databases

“…7,8 More recently, Telik, Inc. (South San Francisco, CA), developed a method for predicting ligand binding to proteins by affinity fingerprinting from a small panel of reference proteins. [9][10][11] After preliminary testing of over 300 proteins from a variety of sources, the authors selected panels of [8][9][10][11][12][13][14][15][16][17][18] proteins that displayed the broadest binding affinities for a set of over 5000 compounds. MLR was used to build "computational surrogates" to predict the binding potencies of additional molecules.…”