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 an analyte in a sample disposed within in a fluid path based on the size of the analyte;
immobilizing the analyte in the fluid path wherein the immobilizing is non-specific to the analyte;
after the immobilizing, conveying a detection reagent through the fluid path; and
detecting the analyte by measuring a signal generated by the detection reagent.
2. The method of claim 1 , wherein the analyte includes a protein.
3. The method of claim 1 , wherein the sample includes a standard.
4. The method of claim 1 , wherein the sample is hydrodynamically introduced into the fluid path.
5. The method of claim 1 , wherein the sample is introduced into the fluid path by electrokinetic injection.
6. The method of claim 1 , wherein the fluid path includes at least one of a capillary, a substrate, a curved surface or a planar surface.
7. The method of claim 1 , wherein the analyte is resolved by electrophoresis.
8. The method of claim 1 , wherein the fluid path includes a sieving matrix.
9. The method of claim 1 , wherein the fluid path includes a detergent.
10. The method of claim 1 , wherein the immobilizing includes immobilizing by photoactivated chemistry.
11. The method of claim 1 , wherein the immobilizing includes heating the fluid path.
12. The method of claim 1 , wherein the detection reagents include antibodies.
13. The method of claim 1 , wherein the resolving includes resolving the analyte prior to the immobilizing to enable the detection of a physical parameter of the analyte.
14. The method of claim 1 , the analyte being one analyte from a plurality of analytes, wherein the immobilizing is non-specific to any analyte from the plurality of analytes.
15. A method, comprising:
resolving in a fluid path an analyte in a sample based on the size of the analyte;
immobilizing in the fluid path the analyte, the immobilizing being one of photo-immobilizing, chemically immobilizing, or thermally immobilizing; and
conveying a detection agent through the fluid path after the immobilizing, the detection agent configured to bind to or interact with the analyte and facilitate detection of the immobilized analyte.
16. The method of claim 15 , wherein the fluid path includes at least one of a capillary, a substrate, a curved surface or a planar surface.
17. The method of claim 15 , the fluid path being a first fluid path from a plurality of fluid paths, the plurality of fluid paths including capillaries.
18. The method of claim 15 , wherein the analyte is resolved by electrophoresis.
19. The method of claim 15 , wherein the analyte is resolved by at least one of isoelectric focusing, gel electrophoresis, or electrokinetic chromatography.
20. The method of claim 15 , the immobilizing including activating a reactive moiety in the fluid path.
21. The method of claim 15 , wherein the detection agent is conveyed through the fluid path by at least one of hydrodynamic flow, electroendosmotic flow, or electrophoresis.
22. The method of claim 15 , wherein the detection agent includes at least one of antibodies, peptides, proteins, enzyme substrates, transition state analogs, cofactors, nucleotides, polynucleotides, aptamers, lectins, small molecules, ligands, inhibitors, drugs, or labeled molecules thereof, chemiluminescent substrates or chemiluminescent labeled antibodies.
23. The method of claim 15 , wherein the resolving and the immobilizing occurs in a same location of the fluid path.
24. The method of claim 15 , 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 resolved analyte when the one or more reactive moieties are activated.
25. A method, comprising:
resolving a protein in a fluid path based on the size of the protein;
photoimmobilizing the resolved protein;
contacting the photoimmobilized protein with an antibody to form an antibody-protein complex in the fluid path; and
detecting the photoimmobilized protein in the fluid path after a detection agent is conveyed through the fluid path.
26. The method of claim 25 , wherein said antibody includes a label.
27. The method of claim 25 , further comprising contacting the antibody-protein complex with a labeled antibody.
28. The method of claim 25 , wherein the detecting includes detecting a chemiluminescent signal.
29. The method of claim 25 , wherein the at least one of the resolved analytes are immobilized in the location of the fluid path during the detecting step.
30. The method of claim 25 , the fluid path including a location, the resolving, the immobilizing and the detecting occurring substantially in the location.
31. The method of claim 25 , the protein being one protein from a plurality of proteins, the photoimmobilizing being non-specific with respect to any the plurality of proteins.
32. The method of claim 25 , wherein the fluid path includes at least one of a capillary, a substrate, a curved surface or a planar surface.