Claims
- 1. A method for detecting the binding of or interaction between each or any of a plurality of ligands and one or more target antiligands, said method comprising:
(a) reacting the antiligand(s) with a set of electrophoretic tag (e-tag) probes, the set comprising j members, and each of said e-tag probes having the form:
(D, Mj)-L-Tj, where
(i) D is a detection group comprising a detectable label; (ii) Tj is a ligand capable of binding to or interacting with a target antiligand, (iii) L is a linking group connected to Tj by a bond that is cleavable by a selected cleaving agent when the probe is bound to or interacting with the target antiligand, wherein cleavage by said agent produces an e-tag reporter of the form (D, Mj)-L′, where L′ is the residue of L attached to (D, Mj) after such cleavage, (iv) Mj is a mobility modifier having a charge/mass ratio that imparts a unique and known electrophoretic mobility to a corresponding e-tag reporter of the form (D, Mj)-L′, within a selected range of electrophoretic mobilities with respect to other e-tag reporters of the same form in the probe set; and (v) (D, Mj)- includes both D-Mj-and Mj-D-; (b) treating the contacted antiligand(s) with the cleaving agent, thereby to produce a mixture of e-tag reporters having the form (D, Mj)-L′, and uncleaved and/or partially cleaved probes, (c) exposing said mixture to a capture agent effective to bind to uncleaved or partially cleaved e-tag probes, but not the corresponding e-tag reporters, and effective to
(i) impart a mobility to the probes bound to capture agent that prevents the probes from electrophoretically migrating within said range of electrophoretic mobilities or (ii) immobilize the probes on a solid support; (d) fractionating e-tag reporters having the form (D, Mj)-L′ by electrophoresis, to effect separation of the e-tag reporters, and (e) identifying the electrophoretic mobilities of one or more electrophoretic bands, each band corresponding to an e-tag reporter that is uniquely assigned to a target antiligand.
- 2. The method of claim 1, wherein Tj is biotinylated and the capture agent is avidin or streptavidin.
- 3. The method of claim 1, wherein Tj contains an antigen and the capture agent is an antibody or antibody fragment that binds specifically to the antigen.
- 4. The method of claim 1, wherein Tj contains a particle or mass group that effectively prevents its migration under electrophoretic conditions within the range of electrophoretic mobilities of the e-tag reporters.
CROSS-REFERENCE TO RELATED APPLICATIONS
[0001] This application is a continuation of 09/303,029 filed Apr. 30, 1999; 09/561,579 filed Apr. 28, 2000; 09/602,586 filed Jun. 21, 2000; 09/684,386 filed Oct. 4, 2000; and 09/698,846 filed Oct. 27, 2000, all of which are incorporated herein by reference in their entirety.
Continuations (5)
|
Number |
Date |
Country |
Parent |
09303029 |
Apr 1999 |
US |
Child |
09824984 |
Apr 2001 |
US |
Parent |
09561579 |
Apr 2000 |
US |
Child |
09824984 |
Apr 2001 |
US |
Parent |
09602586 |
Jun 2000 |
US |
Child |
09824984 |
Apr 2001 |
US |
Parent |
09684386 |
Oct 2000 |
US |
Child |
09824984 |
Apr 2001 |
US |
Parent |
09698846 |
Oct 2000 |
US |
Child |
09824984 |
Apr 2001 |
US |