Real time microarrays
First Claim
1. A method of measuring at least one parameter of a binding reaction of an analyte with a probe on a microarray, comprising the steps of:
- contacting a fluid volume comprising an initial concentration C of at least one analyte that is labeled with a quencher with a solid substrate comprising a number Po of unbound molecules of at least one probe that has been labeled with a donor and affixed thereto at a probe spot, said at least one probe and said at least one analyte configured to bind together, wherein the at least one analyte is not labeled with a metal cluster;
measuring kinetically in real-time a plural number of values for the number P(t) of unbound molecules of said probe at time t, during a time interval when said reaction of said at least one probe and said at least one analyte is occurring, wherein said reaction is not competitive;
determining by using said measured plural number of values P(t) fitted to an exponential curve at least one parameter descriptive of a binding reaction of said at least one probe and said at least one analyte; and
performing a selected one of storing, manipulating and displaying to a user data comprising said at least one parameter,wherein said contacting and measuring are performed in the absence of a washing step.
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Abstract
This invention provides methods and systems for measuring the binding of analytes in solution to probes bound to surfaces in real-time. The method involves contacting a fluid volume having a plurality of different analytes with a solid substrate having a plurality of different probes. The probes are capable of specifically binding to the analytes. The method also involves measuring signals at multiple time points while the fluid volume is in contact with the substrate. The signals measured at multiple time points can be correlated with the amount of binding of the analytes with the probes. The method eliminates the need to wash the probes before measuring the binding characteristics.
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Citations
10 Claims
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1. A method of measuring at least one parameter of a binding reaction of an analyte with a probe on a microarray, comprising the steps of:
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contacting a fluid volume comprising an initial concentration C of at least one analyte that is labeled with a quencher with a solid substrate comprising a number Po of unbound molecules of at least one probe that has been labeled with a donor and affixed thereto at a probe spot, said at least one probe and said at least one analyte configured to bind together, wherein the at least one analyte is not labeled with a metal cluster; measuring kinetically in real-time a plural number of values for the number P(t) of unbound molecules of said probe at time t, during a time interval when said reaction of said at least one probe and said at least one analyte is occurring, wherein said reaction is not competitive; determining by using said measured plural number of values P(t) fitted to an exponential curve at least one parameter descriptive of a binding reaction of said at least one probe and said at least one analyte; and performing a selected one of storing, manipulating and displaying to a user data comprising said at least one parameter, wherein said contacting and measuring are performed in the absence of a washing step. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10)
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Specification