Biomap characterization of biologically active agents
AssigneeBioseek, Inc.
InventorsEllen L. Berg, Eugene C. Butcher, Jennifer Melrose
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Abstract
A method of screening biologically active agent based on the analysis of complex biological responses in culture. Methods for selecting cells and culture conditions for such screens are provided, as well as the identification of an optimized set of discrete parameters to be measured, and the use of biomap analysis for rapid identification and characterization of drug candidates, genetic sequences acting pathways, and the like. A feature of the invention is simultaneous screening of a large number of cellular pathways, and the rapid identification of compounds that cause cellular responses.
Claims16
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-α, TNF-β, 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-γ 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.
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 endothelial 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-α, TNF-β, 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-γ 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 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.
Dependencies are inferred from the claim text.