Methods and compositions for enhancing immunoassays

Patent metadata

Field	Value
Publication number	US-9983206-B2
Application number	US-201414209746-A
Country	US
Kind code	B2
Filing date	Mar 13, 2014
Priority date	Mar 15, 2013
Publication date	May 29, 2018
Grant date	May 29, 2018

Abstract

Embodiments of the methods, compositions, and systems provided herein relate to enzymatic enhancement of immunoassays using photonic sensor arrays.

First claim

What is claimed is:

1. A method of detecting interleukin-2 (IL-2) comprising:
(a) obtaining a fluidic cell comprising an optical ring resonator having a first anti-cytokine antibody attached thereto, wherein the first anti-cytokine antibody is anti-IL-2 from clone 555051 antibody;
(b) contacting the first anti-cytokine antibody with a sample comprising IL-2 that selectively binds to the anti-cytokine antibody;
(c) contacting the bound IL-2 with a second anti-cytokine antibody that selectively binds to the IL-2, wherein the second anti-cytokine antibody is a biotinylated anti-IL-2 from clone 555040 antibody;
(d) contacting the biotinylated second anti-cytokine antibody with a streptavidin-horseradish peroxidase conjugate;
(e) contacting the horseradish peroxidase with a substantially continuous flow of a solution comprising 4-chloro-1-naphthol and hydrogen peroxide, under conditions that oxidize 4-chloro-1-naphthol to 4-chloro-1-naphthon, whereby the 4-chloro-1-naphthon precipitates on the surface of the optical ring resonator during the substantially continuous flow of the solution; and
(f) measuring a change in resonance wavelengths of the optical ring resonator, thereby indicating the presence of the IL-2.

2. The method of claim 1 , wherein the concentration of hydrogen peroxide is less than about 0.003%.

3. The method of claim 1 , wherein a concentration of the cytokine less than about 100 pg/ml is detected.

4. The method of claim 1 , wherein a continuous flow of reagents contacting the optical ring resonator having a first anti-cytokine antibody attached thereto is maintained for step (b).

5. The method of claim 1 , wherein a substantially continuous flow of reagents contacting the optical ring resonator having a first anti-cytokine antibody attached thereto is maintained for each of steps (b), (c), (d), and (e).

6. The method of claim 1 , wherein a continuous flow of reagents contacting the optical ring resonator having a first anti-cytokine antibody attached thereto is maintained for each of steps (b), (c), (d), and (e).

7. The method of claim 1 , wherein steps (b)-(e) comprise maintaining a flow of reagents contacting the optical ring resonator having a first anti-cytokine antibody attached thereto.

8. The method of claim 7 , wherein the flow rate of the reagents is about 30 μL/min.

9. The method of claim 1 , wherein a continuous flow of the solution is maintained for step (e).

10. The method of claim 9 , wherein the continuous flow of the solution is maintained for steps (e) and (f).

Assignees

• Univ Illinois

Inventors

• Bailey Ryan C
• Luchansky Matthew S
• Qavi Abraham J
• Kindt Jared T

Classifications

• G01N33/54373 Primary

  involving physiochemical end-point determination, e.g. wave-guides, FETS, gratings · CPC title

• G01N2333/5412

  IL-6 · CPC title

• G01N2333/55

  IL-2 · CPC title