Methods for detecting analytes
View Patent ↗The present disclosure provides methods for the detection of multiple analytes using a single solid phase. The present disclosure also relates to the preparation of solid phases that include receptacles having affixed thereto antibodies directed to at least two different analytes. The methods of the present disclosure can be used, for example, for the quantitative detection of analytes in a sample and the measurement thereof.
1. A method for detecting analytes comprising:
providing a solid phase comprising two receptacles, wherein each of said two receptacles comprises a first capture antibody and a second capture antibody affixed thereto, wherein the first capture antibody binds to a first analyte and the second capture antibody binds to a second analyte, and wherein said first analyte and said second analyte are different;
providing a sample, wherein a portion of the sample is provided to each of the two receptacles;
providing to the solid phase, a first detection antibody specific to the first analyte and a second detection antibody specific to the second analyte, wherein the portion of said sample in the first receptacle is contacted with said first detection antibody and the portion of said sample in the second receptacle is contacted with said second detection antibody, and wherein said first detection antibody and said second detection antibody produce the same detectable signal; and
detecting said first analyte and detecting said second analyte in said sample.
2. The method of claim 1 , wherein said two receptacles are tapered receptacles.
3. The method of claim 1 , further comprising quantifying the amount of analyte detected in each of said two receptacles.
4. The method of claim 3 , wherein said quantifying comprises obtaining the detected amount of analyte for each receptacle and providing a single numerical value.
5. The method of claim 1 , further comprising dispensing in each of said two receptacles a luminogenic substrate and an electron transfer agent, wherein said luminogenic substrate produces the detectable signal in each of said first receptacle and said second receptacle.
6. The method of claim 5 , wherein said detectable signal in each of said first receptacle and said second receptacle is light.
7. The method of claim 6 , wherein said detectable signal in each of said first receptacle and said second receptacle is the same wavelength of light.
8. The method of claim 7 , wherein said amount of said first analyte in said sample is directly proportional to the amount of light detected in said first receptacle.
9. The method of claim 8 , wherein said amount of said second analyte in said sample is directly proportional to the amount of light detected in said second receptacle.
10. The method of claim 5 , wherein said luminogenic substrate is oxidized by contacting said detection antibody in the first receptacle or by contacting said detection antibody in the second receptacle.
11. The method of claim 5 , wherein said luminogenic substrate comprises a luminol derivative and peracid salt.
12. The method of claim 5 , wherein said electron transfer agent is a substituted acetanilide.
13. The method of claim 1 , wherein said first detection antibody and said second detection antibody are each conjugated to horseradish peroxidase.
14. The method of claim 1 , further comprising incubating the portion of said sample in the first receptacle with the first detection antibody and the portion of said sample in the second receptacle with the second detection antibody for at least five minutes prior to said detecting the amount of each of said first analyte in said sample and said second analyte in said sample.
15. The method of claim 14 , wherein said incubation is for a duration of 8 minutes.
16. The method of claim 1 , wherein said amount of the first analyte in said sample and said amount of the second analyte in said sample are detected in succession.
17. The method of claim 1 , wherein said sample, and said first detection antibody specific to the first analyte and said second detection antibody specific to the second analyte are provided to said solid phase at the same time.
18. The method of claim 1 , wherein said sample is a body fluid sample.
19. The method of claim 18 , wherein said body fluid sample is urine.
20. The method of claim 1 , wherein each of said first capture antibody, said second capture antibody, said first detection antibody and said second detection antibody are monoclonal antibodies.
21. The method of claim 1 , wherein said solid phase is configured for use in an automated immunodiagnostic device.
22. The method of claim 21 , wherein said portion of said sample is provided to each of the two receptacles by an automated immunodiagnostic device.
23. The method of claim 22 , wherein said automated immunodiagnostic device provides said first detection antibody to the portion of the sample in the first receptacle and provides said second detection antibody to the portion of the sample in the second receptacle.
24. The method of claim 23 , wherein said detection comprises measurement of an amount of each of said first analyte in said sample and detecting an amount of said second analyte in said sample by said automated immunodiagnostic device.
25. The method of claim 1 , wherein said detecting each analyte comprises providing said solid phase to an automated immunodiagnostic device, wherein said automated immunodiagnostic device detects said first analyte in said sample in said first receptacle and detects said second analyte in said sample in said second receptacle.
26. The method of claim 25 , wherein said automated immunodiagnostic device comprises a luminometer, and wherein said luminometer detects each of said first analyte said second analyte in succession.