Claims
- 1. A method for testing an agent for its potential to inhibit heparanase catalytic activity, comprising the steps of:
interacting immobilized heparanase substrate binding protein on a solid support with labeled heparanase substrate to provide immobilized labeled heparanase substrate, interacting a heparanase enzyme solution with the immobilized labeled heparanase substrate in the presence of the agent, detecting the presence or absence of label in the solution remote from the solid support, to determine whether or not the agent has the potential to inhibit heparanase.
- 2. The method according to claim 1, wherein the label of the labeled heparanase substrate is at least one molecule chosen from fluorescent, radioactive, chemiluminescent, and chromogenic molecules.
- 3. The method according to claim 2, wherein the fluorophores comprise fluorescein, lanthanide-chelate, and Eu(DPTA)-chelate.
- 4. The method according to claim 3, wherein the fluorescein is selected from FITC and F2FITC.
- 5. The method according to claim 3, wherein the lanthanide-chelate is selected from europium-chelate (Eu-chelate), samarium-chelate (Sm-chelate), terbium-chelate (Tb-chelate) and dysprosium-chelate (Dy-chelate).
- 6. The method according to claim 5, wherein the lanthanide-chelate is Eu-chelate.
- 7. The method according to claim 1, wherein the heparanase substrate of the labeled heparanase substrate is selected from heparan sulfate, heparan sulfate proteoglycan and heparan sulfate analogs.
- 8. The method according to claim 7, wherein the heparanase substrate is heparan sulfate.
- 9. The method according to claim 1, wherein the heparanase substrate binding protein is selected from fibroblast growth factor (FGF), VEGF, and PDGF.
- 10. The method according to claim 9, wherein the heparanase substrate binding protein is FGF.
- 11. A method for testing an agent for its potential to inhibit heparanase catalytic activity, comprising the steps of:
interacting immobilized molecules on a solid support with labeled heparanase substrate bound to an immobilization bridge, wherein the immobilized molecules bind to the immobilization bridge, to provide bridge-immobilized labeled heparanase substrate, interacting a heparanase enzyme solution with the bridge-immobilized labeled heparanase substrate in the presence of the agent, detecting the presence or absence of label in the solution remote from the solid support, to determine whether or not the agent has the potential to inhibit heparanase.
- 12. The method according to claim 11, wherein the immobilization bridge is biotin and the immobilized molecules are selected from streptavidin and avidin molecules and mixtures thereof.
- 13. The method according to claim 12, wherein the immobilized molecules are streptavidin molecules.
- 14. A method for testing an agent for its ability to inhibit binding between fibroblast growth factor (FGF) and heparanase substrate, comprising the steps of:
interacting in solution immobilized FGF on a solid support with the agent and labeled heparanase substrate, and detecting the presence or absence of label in the solution remote from the solid support, to determine whether or not the agent has the ability to inhibit binding between fibroblast growth factor and heparanase substrate.
PRIORITY TO PROVISIONAL APPLICATION(S) UNDER 35 U.S.C. §119(e)
[0001] This application claims priority under 35 U.S.C. §119(e) of provisional application(s) Serial No. 60/341,573, filed Dec. 18, 2001.
Provisional Applications (1)
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Number |
Date |
Country |
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60341573 |
Dec 2001 |
US |