Function homology screening
AssigneeBioseek, Inc.
InventorsEllen L. Berg, Eugene C. Butcher, Jennifer Melrose
Life of this patent
Estimating the term…
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 screening system for determining the effect of a candidate compound on inflammatory response, said system comprising a test biological culture, assay reagents for measuring at least two parameters of ICAM-1, VCAM-1 E-selectin, P-selectin, IL-8, CD31, HLA-DR, eotaxin-3, MCP-1 MIG;
- and a data processor for analyzing the data from said biological culture in relation to at least one control biological culture of known activity, said biological culture comprising:
- endothelial cells, at least two factors of TNF-α, IL-1, IL-4, histamine and IFN-γ in amounts sufficient to induce an inflammatory state for said endothelial cells.
A method for testing the activity of a candidate compound on inflammatory response, said method comprising:
- adding a candidate compound to a test biological culture comprising endothelial cells in an inflammatory state as a result of adding to said culture at least two of TNF-α, IL-1, IL-4, histamine and IFN-γ in an amount and incubating for a time sufficient to induce said inflammatory state;
- measuring at least two parameters of ICAM-1, VCAM-1 E-selectin, P-selectin, IL-8, CD31, HLA-DR, eotaxin-3, MCP-1, and MIG and comparing the measurement of said at least two parameters with the measurement from a control biological culture,
- whereby the activity of said candidate compound is determined.
A method for testing the activity of a candidate compound on inflammatory response, said method comprising:
- adding a candidate compound to a test biological culture comprising endothelial cells in an inflammatory state as a result of adding to said culture the factors TNF-α, IL-1, IL-4, histamine and IFN-γ in an amount and incubating for from about 6 to 72h sufficient to induce said inflammatory state;
- measuring at least four parameters of ICAM-1, VCAM-1, E-selectin, P-selectin, eotaxin-3, MCP-1, IL-8, CD31, HLA-DR and MIG and comparing the measurement of said at least four parameters with the measurement from a control biological culture,
- whereby the activity of said candidate compound is determined.
A method for analyzing which cellular pathway of cells in an inflammatory state is affected by a compound, said method comprising:
- adding a candidate compound to one portion of a test biological culture comprising endothelial cells in an inflammatory state as a result of adding to said culture at least two of the factors TNF-α, IL-1, IL-4, histamine and IFN-γ in an amount and incubating for a time sufficient to induce said inflammatory state, and adding an inhibitor compound known to inhibit one pathway to both of said portions;
- measuring at least two parameters of ICAM-1, VCAM-1 E-selectin, P-selectin, IL-8, CD31, HLA-DR, eotaxin-3, MCP-1, and MIG in each of said portions and comparing the measurement of said at least two parameters in each of said portions;
- whereby when said compound has no effect on said parameters in the portion containing said compound as compared to said portion in which said compound is present, said compound is affecting the same pathway as said inhibitor compound.
Dependencies are inferred from the claim text.