Methods for detecting an analyte in a sample
View Patent ↗Methods for detecting the presence of an analyte in a sample are provided. Aspects of the methods include mixing a sample with an indirectly-detectable labeled binding member that includes a label moiety. The mixing produces a first composition. Unbound labeled binding member is then separated from any resultant binding complexes to produce a second composition that includes the binding complexes. A second light emitted by a light wavelength converter that is excited by a first light generated by the labeled binding member of the binding complexes of the second composition is then detected to detect the presence of the analyte in the sample. Embodiments also include kits and systems that find use in practicing the subject methods.
1. A method of detecting the presence of an analyte in a sample, said method comprising:
mixing said sample with a chemiluminescent indirectly-detectable labeled binding member comprising a label moiety to produce a first composition that includes binding complexes comprising said labeled binding member;
separating said unbound labeled binding member from binding complexes present in said first composition to produce a second composition comprising said binding complexes;
adding a light wavelength converter and an oxidant to the second composition;
detecting said binding complexes in said second composition to detect the presence of said analyte in said sample, wherein said detecting comprises detecting second light emitted by said light wavelength converter, said light wavelength converter generating said second light in response to absorbing first light generated by said indirectly-detectable labeled binding member of said binding complexes of said second composition, said first light being generated in response to reaction of said indirectly-detectable labeled binding member with said oxidant.
2. The method according to claim 1 , wherein said binding complexes are present in said second composition in amount proportional to the amount of said analyte in said sample.
3. The method according to claim 1 , wherein said labeled binding member specifically binds to said analyte.
4. The method according to claim 3 , wherein said first composition further comprises a second binding member, and said binding complexes are made up of said labeled binding member, said analyte and said second binding member.
5. The method according to claim 4 , wherein said second binding member is immobilized on a surface of a solid support.
6. The method according to claim 1 , wherein said labeled binding member does not specifically bind to said analyte.
7. The method according claim 6 , wherein said labeled binding member competes with said analyte for binding to a second binding member.
8. The method according to claim 1 , wherein said light wavelength converter is at least one of linked directly to said labeled binding member and linked to said labeled binding member via a linker moiety.
9. The method according to claim 8 , wherein said light wavelength converter is conjugated to said labeled binding member.
10. The method according to claim 8 , wherein said light wavelength converter is immobilized on a surface of a solid support.
11. The method according to claim 1 , wherein said light wavelength converter remains unlinked with said labeled binding member in said second composition.
12. The method according to claim 1 , wherein said label moiety of said indirectly-detectable labeled binding member is an enzymatic label.
13. The method according to claim 12 , wherein said enzymatic label comprises a peroxidase.
14. The method according to claim 12 , wherein said enzymatic label converts a substrate into a chemiluminescent product.
15. The method according to claim 14 , wherein said substrate is a dihydrophthalazinedione.
16. The method according to claim 15 , wherein said substrate is luminol.
17. The method according to claim 1 , wherein said light wavelength converter has a Stokes shift of at least about 100 nm.
18. The method according to claim 17 , wherein said light wavelength converter comprises a nanoparticle.