Methods and devices for analyte detection
View Patent ↗Methods for detecting one or more analytes, such as a protein, in a fluid path are provided. The methods include resolving, immobilizing and detecting one or more analytes in a fluid path, such as a capillary. Also included are devices and kits for performing such assays.
1. A method, comprising:
resolving in a fluid path one or more analytes in a sample;
immobilizing in the fluid path at least one of the one or more resolved analytes, the immobilizing being non-specific to any analyte wherein the immobilizing being at least one of photo-immobilizing, chemically immobilizing, or thermally immobilizing; and
conveying detection agents through the fluid path after the immobilizing, the detection agents configured to bind to or interact with the analytes and facilitate detection of at least one of the immobilized analytes.
2. The method of claim 1 , wherein the fluid path comprises a capillary.
3. The method of claim 1 , further comprising a plurality of fluid paths, wherein the plurality of fluid paths are capillaries.
4. The method of claim 1 , wherein the one or more analytes are resolved by electrophoresis.
5. The method of claim 1 , wherein the one or more analytes are resolved by isoelectric focusing, gel electrophoresis, or electrokinetic chromatography.
6. The method of claim 1 , wherein the fluid path includes one or more reactive moieties capable of immobilizing the at least one of the one or more analytes in said fluid path, the immobilizing including activating at least one of the one or more reactive moieties.
7. The method of claim 6 , wherein the activating comprises photoactivation, chemical activation or thermal activation of the activated reactive moieties.
8. The method of claim 1 , wherein the detection agents are conveyed through the fluid path by hydrodynamic flow, electroendosmotic flow, or electrophoresis.
9. The method of claim 1 , wherein said detection agents comprise antibodies, peptides, proteins, enzyme substrates, transition state analogs, co factors, nucleotides, polynucleotides, aptamers, lectins, small molecules, ligands, inhibitors, drugs, or labeled molecules thereof, or chemiluminescent substrates.
10. The method of claim 1 , wherein said detection agents comprise chemiluminescent labeled antibodies.
11. The method of claim 1 , wherein the detection of the at least one of the immobilized analytes includes detecting a chemiluminescent signal.
12. The method of claim 1 , wherein the detection of the at least one of the immobilized analytes is by a photodetector.
13. The method of claim 1 , wherein the detection of the at least one of the immobilized analytes is by an array of photodetectors.
14. The method of claim 1 , wherein the detection of the at least one of the immobilized analytes is by a charged coupled device (CCD) array.
15. The method of claim 1 , wherein the resolving and the immobilizing occurs in a same location of the fluid path.
16. The method of claim 1 , wherein one or more reactive moieties are disposed within the fluid path prior to the resolving, the one or more reactive moieties configured to immobilize the at least one of the resolved analytes when the one or more reactive moieties are activated.
17. A method, comprising:
resolving one or more proteins at a location in a capillary;
photoimmobilizing at least one of the resolved proteins at the location in the capillary;
contacting the at least one of the photoimmobilized proteins with antibodies to form one or more antibody-protein complexes in the capillary; and
detecting at least one of the at least one of the photoimmobilized proteins at the location in the capillary after one or more detection agents are conveyed through the capillary.
18. The method of claim 17 , wherein said antibodies comprise a label.
19. The method of claim 17 , further comprising contacting said one or more antibody-protein complexes with a labeled antibody.
20. The method of claim 17 , wherein said detecting includes detecting a chemiluminescent signal.
21. The method of claim 17 , wherein the photoimmobilizing is non-specific with respect to any of the one or more resolved proteins.
22. A method, comprising:
resolving one or more proteins in a location of a capillary, the capillary having one or more photoreactive groups disposed therein;
photoimmobilizing at least one of the resolved proteins in the location of the capillary;
introducing antibodies into the capillary such that the antibodies contact the photoimmobilized proteins to form one or more antibody-target complexes in the location of the capillary; and
detecting in the location of the capillary at least one of the photoimmobilized proteins after one or more detection agents are conveyed through the capillary and contact the one or more antibody-target complexes in the location of the capillary.
23. The method of claim 22 , wherein the photoimmobilizing is non-specific with respect to any of the one or more resolved proteins.
24. The method of claim 22 , wherein the capillary has disposed therein one or more photoreactive groups and one of polymeric materials or polymerizable materials including one or more photoreactive groups.
25. A method, comprising:
concentrating one or more proteins in a fluid path;
immobilizing at least one of the concentrated proteins in the fluid path, the immobilization being non-specific with respect to any of the proteins wherein the immobilizing being at least one of photo-immobilizing, chemically immobilizing, or thermally immobilizing;
introducing antibodies into the fluid path such that the immobilized proteins contact the antibodies to form one or more antibody-protein complexes in the fluid path; and
after the introducing, detecting at least one of the immobilized proteins.
26. The method of claim 25 , wherein the immobilizing and the detecting occurs in a same location of the fluid path.
27. A method, comprising:
resolving one or more analytes in a location of a fluid path wherein the immobilizing being at least one of photo-immobilizing, chemically immobilizing, or thermally immobilizing;
immobilizing at least one of the resolved analytes in the location of the fluid path;
after the immobilizing, conveying detection agents through the fluid path, the detection agents configured to bind to or interact with the immobilized analytes after the detection agents are conveyed through the fluid path, the detection agents configured to facilitate detection of the immobilized analytes; and
detecting at least one of the immobilized analytes.
28. The method of claim 27 , wherein the at least one of the resolved analytes are immobilized in the location of the fluid path during the detecting step.
29. The method of claim 27 , wherein the detecting occurs in the location of the fluid path.
30. The method of 27 , wherein said immobilizing step and said detecting step are carried out by a same device.
31. The method of claim 27 , wherein the immobilizing is non-specific with respect to any of the one or more resolved analytes.