Capillary assays involving separation of free and bound species
First Claim
1. A method for detecting the presence of an analyte in a sample employing capillary electroseparation in a microfluidic device where the binding of specific binding pair members results in an inhomogeneous complex, said method comprising:
- combining in an assay mixture particles of uniform dimension to which one member of said specific binding pair is bound, said sample suspected of containing said analyte and a labeled compound which binds to said bound member of said specific binding pair or to said analyte;
subjecting said assay mixture to capillary electroseparation to provide a sharp band of particles; and
detecting labeled member bound to said particles, as indicative of the presence of said analyte.
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Abstract
The present invention concerns methods for masking inhomogeneity of a member of a specific binding pair (sbp) employed in a capillary electroseparation. The method comprises binding the sbp member to synthetic particles that become localized during capillary electroseparation. Also disclosed is one embodiment of the present invention, which is a method for conducting a capillary electroseparation specific binding assay. The method involves the electroseparation of a labeled first member of a specific binding pair that is bound in a complex from labeled first member that is not bound in the complex. The complex comprises the first member and a second member of a specific binding pair. A combination is provided comprising a sample suspected of containing an analyte, a labeled first member of a specific binding pair, and a second member of a specific binding pair under conditions for forming a complex between labeled first member and the second member. The second member either initially or subsequent to the formation of the complex being covalently or noncovalently bound to synthetic particles that migrate uniformly during electroseparation. The combination is subjected to electroseparation and a determination is made as to whether the complex is formed. Also disclosed are kits for conducting a capillary electroseparation specific binding assay.
473 Citations
10 Claims
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1. A method for detecting the presence of an analyte in a sample employing capillary electroseparation in a microfluidic device where the binding of specific binding pair members results in an inhomogeneous complex, said method comprising:
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combining in an assay mixture particles of uniform dimension to which one member of said specific binding pair is bound, said sample suspected of containing said analyte and a labeled compound which binds to said bound member of said specific binding pair or to said analyte; subjecting said assay mixture to capillary electroseparation to provide a sharp band of particles; and detecting labeled member bound to said particles, as indicative of the presence of said analyte. - View Dependent Claims (2, 3, 4, 5, 6)
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7. A method for conducting a capillary electroseparation specific binding assay for a protein analyte using a microfluidic device, said method involving the electroseparation of a labeled first member of a specific binding pair, where the other member is said analyte, and said analyte becomes bound to a second specific binding pair member, that binds to analyte in the presence of binding of said analyte to said first member, said method comprising:
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(a) incubating an assay mixture comprising a sample suspected of containing said analyte, said first member bound to particles of uniform dimension, and said second member in any order of addition, to form a complex, when said analyte is present, of said first member bound to said particles, said analyte, and said second member; (b) electroseparating said assay mixture; and (c) determining whether said complex is formed. - View Dependent Claims (8, 9, 10)
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Specification