Rapid Detection of Anti-Chromatin Autoantibodies in Human Serum using a Portable Electrochemical Biosensor

• US 20090288960A1
• Filed: 02/12/2009
• Published: 11/26/2009
• Est. Priority Date: 02/12/2008
• Status: Abandoned Application

• Current Assignee: Gabriel Lopez, Konstantin Konstantinov, Plamen Atanassov, Ravil Sitdikov, Robert Rubin
• Original Assignee: Gabriel Lopez, Konstantin Konstantinov, Plamen Atanassov, Ravil Sitdikov, Robert Rubin
• Inventors: Rubin, Robert, Konstantinov, Konstantin, Lopez, Gabriel, Atanassov, Plamen, Sitdikov, Ravil
• Application Number:
  US12/370,403
• Publication Number:
  US 20090288960A1
• Time in Patent Office: Days
• US Class Current: 205/777.500
• CPC Class Codes: G01N 2800/104 Lupus erythematosus [SLE]
• CPC Class Codes: G01N 33/5438 Electrodes
• CPC Class Codes: G01N 33/564 for pre-existing immune complex or autoimmune disease , i.e. systemic lupus erythematosus, rheumatoid arthritis, multiple sclerosis, rheumatoid factors or complement components C1-C9

Abstract

A rapid electrochemical flow-through sensor and method for detecting the presence of autoantibodies in a fluid sample such as blood is shown and described. The sensor may take either a single- or multi-plexed form. The sensor comprises a fluid inlet, a reaction region comprising immobilized autoantigens and an electrode assembly, and a fluid outlet.

Claims

1. A method for detecting autoantibodies associated with an autoimmune disease in a human test sample comprising:

providing an electrochemical flow-through sensor comprising;

an inlet channel configured to deliver fluid into a reaction region;

a reaction region comprising;

an electrode assembly comprising a working electrode; and

immobilized autoantigens configured to present the product of an enzymatic reaction to the electrode assembly so as to produce a detectable alteration in the working electrode'"'"'s signal output; and

an outlet channel configured to allow for fluid flow out of the reaction region, wherein the outlet channel is not the inlet channel;

introducing the test sample to the reaction region via the inlet channel under sufficient conditions for autoantibodies in the test sample to bind to the immobilized autoantigens;

allowing unbound molecules to exit the reaction region via the outlet channel;

introducing one or more detection molecules configured to, upon interaction with the bound autoantibodies, produce an enzymatic reaction, the product of which is able to produce a detectable alteration in the working electrode'"'"'s signal output.

2. The method of claim 1 wherein the autoimmune disease is systemic lupus erythematosus.

3. The method of claim 1 wherein the autoantibodies are anti-chromatin autoantibodies.

4. The method of claim 1 wherein the immobilized autoantigens is H1-stripped chromatin.

5. The method of claim 4 wherein the human test sample is blood.

6. The method of claim 1 wherein the outlet channel runs through the working electrode.

7. A flow-through electrochemical apparatus for detecting autoantibodies associated with an autoimmune disease in a human test sample, the apparatus comprising:

a housing containing;

an inlet channel configured to deliver fluid into a reaction region;

a reaction region comprising;

an electrode assembly comprising a working electrode and a reference; and

immobilized autoantigens configured to present the product of an enzymatic reaction to the electrode assembly so as to produce a detectable alteration in the working electrode'"'"'s signal output; and

an outlet channel configured to allow for fluid flow out of the reaction region, wherein the outlet channel is not the inlet channel.

8. The apparatus of claim 7 wherein the autoantigens are immobilized to a porous substrate.

9. The apparatus of claim 7 wherein the electrode assembly comprises at least one working electrode and at least once reference electrode.

10. The apparatus of claim 9 wherein the electrode assembly comprises a substrate.

11. The apparatus of claim 10 wherein the working and reference electrodes are screen-printed on a paper substrate.

12. The apparatus of claim 7 wherein the outlet channel travels through the working electrode.

13. The apparatus of claim 7 wherein the autoantigens are H1-stripped chromatin.

14. The apparatus of claim 7 further comprising a second inlet channel configured to deliver fluid into a second reaction region.

15. A flow-through electrochemical apparatus for detecting autoantibodies associated with an autoimmune disease in a human test sample, the apparatus comprising:

a housing containing;

a plurality of inlet channel configured to deliver fluid into a plurality of reaction regions, wherein each reaction region comprises;

an electrode assembly comprising a working electrode and a reference; and

immobilized autoantigens configured to present the product of an enzymatic reaction to the electrode assembly so as to produce a detectable alteration in the working electrode'"'"'s signal output; and

a plurality of outlet channels configured to allow for fluid flow out of the reaction regions, wherein the outlet channels are not the inlet channels.

16. The apparatus of claim 15 wherein the plurality of reaction regions have the same autoantigens immobilized therein.

17. The apparatus of claim 16 wherein apparatus comprises a single porous substrate having the autoantigens immobilized thereto;

and wherein the porous substrate is in fluid contact with each of the plurality of inlet channels.

18. The apparatus of claim 16 wherein the autoantigens are H1-stripped chromatin.

19. The apparatus of claim 15 wherein the apparatus contains a single substrate including a plurality of electrode assemblies and wherein each inlet channel delivers fluid to a different electrode assembly on the single substrate.

20. The apparatus of claim 15 wherein at least two of the reaction regions have different autoantigens immobilized therein.