Patent ID: 6524866
Filing Date: 2003-02-25
Classification: G01N

Abstract:
A method to detect, in a sample of complex biological material, any candidate hit compound that binds to a selected target at or above a selected binding strength and to determine a stability, during capillary electrophoresis, of any target/hit compound complex detected, wherein the stability is determined to identify a relative affinity of a hit compound, said method comprising, in the order given, the steps of:(1) providing a predetermined concentration of a selected target; (2) providing a sample of complex biological material; (3) mixing the target with the sample to form a target/sample mixture and incubating the target/sample mixture; (4) injecting an aliquot of the target/sample mixture into a capillary electrophoresis instrument comprising a capillary having a single detection point; (5) subjecting the target/sample aliquot to capillary electrophoresis under a set of conditions to detect any hit compound in the sample that binds to the target at or above a selected binding strength; (6) tracking a capillary electrophoretic migration of the target in the injected target/sample aliquot at the detection point in the capillary electrophoresis instrument; (7) repeating steps (4)-(6) one or more times, wherein each performance of steps (5)-(6) uses a different set of conditions for a different selected binding strength, wherein each injected target/sample aliquot undergoes a different capillary electrophoresis run time from other injected target/sample aliquot(s), thereby generating multiple capillary electrophoretic profiles, of the same target/sample aliquot, wherein the different conditions are optimized so that any complex, between the target and any candidate hit compound at or above each selected binding strength, generates a different result in tracking the capillary electrophoretic migration as in step (6); (8) determining whether at least one capillary electrophoretic profile of the target generated from step (6), when compared to a reference standard, indicates a presence of any hit compound(s) binding to the target; and (9) determining the stability, during capillary electrophoresis, of any target/hit compound complex(es) detected, by comparing the multiple capillary electrophoretic profiles.