Patent ID: 6656695
Filing Date: 2003-12-02
Classification: C12Q,G01N

Abstract:
A method for determining the activity of a biologically active agent according to its effect on cellular signaling pathways, the method comprising:contacting a candidate biologically active agent with a cell culture assay combination, wherein said cell culture assay combination comprises human cells in an inflammatory state as a result of adding to said culture at least three of the factors selected from the group consisting of TNF-&agr;, TNF-&bgr;, IL-1, IL-2, IL-4, IL-12, IL-13, Staphylococcal Enterotoxin B (SEB); Staphylococcal Enterotoxin E (SEE), toxic shock syndrome toxin (TSST), anti-CD3 antibody, anti-T cell receptor antibody, histamine and IFN-&ggr; in an amount and for a time sufficient to induce said inflammatory state; recording changes in at least two different cellular parameter readouts, said parameters being selected from the group consisting of ICAM-1, VCAM-1, E-selectin, P-selectin, IL-8, CD31, CD40, HLA-DR, eotaxin-3, MCP-1, and MIG, after introduction of said agent; deriving a biomap dataset from said changes in parameter readouts wherein said biomap comprises data normalized to control data on the same cell type under control conditions comprising said at least three factors and lacking said biologically active agent, and wherein output parameters are optimized so that the biomap dataset is sufficiently informative that it can discriminate the mode of action or functional effect of an agent; comparing said biomap dataset to a reference biomap dataset to determine the presence of variation, wherein the presence of variation indicates a difference in the effect of the agent on a cellular signaling pathway.