TEST APPARATUS AND METHODS FOR ST2 CARDIAC BIOMARKER

• US 20160169879A1
• Filed: 12/11/2014
• Published: 06/16/2016
• Est. Priority Date: 12/11/2014
• Status: Active Grant

• Current Assignee: Critical Care Diagnostics Inc.
• Original Assignee: Critical Care Diagnostics Inc.
• Inventors: Snider, James V., Bender, Jillian Elizabeth, Kuo, Shrin P., Chung, Roy A.
• Granted Patent: US 10,324,089 B2
• Time in Patent Office: Days
• US Class Current: 1/1
• CPC Class Codes: G01N 2333/7155 for interleukins [IL]
• CPC Class Codes: G01N 2800/324 Coronary artery diseases, e.g. angina pectoris, myocardial infarction
• CPC Class Codes: G01N 2800/325 Heart failure or cardiac arrest, e.g. cardiomyopathy, congestive heart failure
• CPC Class Codes: G01N 33/54388 based on lateral flow
• CPC Class Codes: G01N 33/558 using diffusion or migration of antigen or antibody immunochromatographic test strips G01N33/54387
• CPC Class Codes: G01N 33/566 using specific carrier or receptor proteins as ligand binding reagents where possible specific carrier or receptor proteins are classified with their target compounds

Abstract

The technology described in this document can be embodied in a test strip for use in measuring a level of an ST2 cardiac biomarker in a whole blood sample. The test strip includes a base, and a plurality of conjugates, wherein each conjugate includes a reporter group bound to a first antibody that binds to ST2. A conjugate pad holds the plurality of conjugates that bind with ST2 to produce conjugate-ST2 complexes. The test strip also includes a plurality of second antibodies that bind to ST2, and a plurality of third antibodies that bind to the conjugate-ST2 complexes. The plurality of second antibodies are bound to a membrane in a test location, and the plurality of third antibodies are bound to the membrane in a control location. A plasma separation pad passes blood plasma from a whole blood sample to the conjugate pad while inhibiting other components.

Claims

1. A test strip for use in measuring a level of an ST2 cardiac biomarker in a whole blood sample, the test strip comprising:

a base;

a plurality of conjugates, each conjugate comprising a reporter group bound to a first antibody that binds to ST2;

a conjugate pad disposed along a length of the base and configured to hold the plurality of conjugates that bind with ST2 to produce conjugate-ST2 complexes;

a plurality of second antibodies that bind to ST2;

a plurality of third antibodies that bind to the conjugate-ST2 complexes;

a membrane disposed on the base such that the membrane is in fluid communication with the conjugate pad, and wherein the plurality of second antibodies are bound to the membrane in a test location and the plurality of third antibodies are bound to the membrane in a control location arranged further from the conjugate pad than the test location; and

a plasma separation pad in fluid communication with a portion of the conjugate pad, wherein the plasma separation pad is disposed and configured to receive the whole blood sample and to pass blood plasma from the whole blood sample to the conjugate pad while inhibiting other components of the whole blood sample from passing to the conjugate pad.

2. The test strip of claim 1, wherein either the first or the second antibodies bind specifically to ST2.

3. The test strip of claim 1, wherein both the first and second antibodies bind specifically to ST2.

4. The test strip of claim 1, further comprising an absorbent pad disposed on the base, and in fluid communication with the membrane at an end or side of the membrane opposite the conjugate pad, wherein the absorbent pad is configured to absorb plasma and buffer that has traversed through the membrane.

5. The test strip of claim 1, wherein the conjugate pad is disposed on the base to receive a buffer solution, and a portion of the conjugate pad that receives the buffer solution is not in contact with the plasma separation pad.

6. The test strip of claim 1, wherein the conjugate pad comprises fibers comprising glass, polyester, or glass and polyester.

7. The test strip of claim 1, wherein at least one of the plasma separation pad or the membrane comprises nitrocellulose.

8. The test strip of claim 1, wherein the first antibodies comprise monoclonal antibodies.

9. The test strip of claim 1, wherein the reporter group comprises gold particles.

10. The test strip of claim 8, wherein the monoclonal antibodies comprise 7E4-monoclonal-anti-ST2 antibodies that are conjugated to colloidal gold.

11. The test strip of claim 1, wherein the second antibodies are 9F8-monoclonal-anti-ST2 antibodies.

12. The test strip of claim 1, wherein the third antibodies comprise goat anti-mouse IgG antibodies.

13. The test strip of claim 1, wherein an amount of change in a visual appearance of a first portion that includes the second antibodies depends on an amount of the bound conjugate present in the blood plasma traversing the first portion.

14. A test apparatus comprising:

a test strip having a first end and an opposite second end; and

a housing for the test strip, wherein the housing comprises;

a first section comprising an outer face and an inner face, wherein the inner face of the first section comprises a channel to receive the test strip along the length of the first section, and

a second section comprising an outer face and an inner face, wherein the second section is configured to be attached to the first section such that in an attached configuration the inner face of the first section faces the inner face of the second section, and the first and second sections together enclose the test strip within the housing, wherein the second section comprises;

a buffer port configured to allow a buffer solution to be dispensed to a portion of the test strip proximate to the first end of the test strip,

a test window configured to provide visualization of one or both of a test location and a control location on the test strip,

a sample port disposed between the buffer port and the test window, the sample port configured to enable a sample of whole blood to be dispensed to the test strip, and

a set of projections disposed on the inner face of the second section between the test window and the sample port such that in the attached configuration, each projection in the set of projections is in contact with the test strip, wherein

a height of at least one projection in the set of projections is different from a height of another projection in the set of projections, and heights of the different projections are configured such that, in the attached configuration, the set of projections produces a pressure gradient that allows a fluid to flow at a predetermined flow rate along the length of the test strip between the first and last projections in the set of projections.

15. The test apparatus of claim 14, wherein the predetermined flow rate is such that the fluid flows from the portion of the test strip adjacent to the buffer port of the second section to the portion of the test strip adjacent to the test window of the second section in about 15 minutes.

16. The test apparatus of claim 14, wherein the set of projections is a set of ridges.

17. The test apparatus of claim 16, wherein distances between ridges are substantially equal.

18. The test apparatus of claim 16, wherein the set of projections includes three ridges, a first ridge being disposed closer to the sample port than the other ridges, a third ridge being disposed closer to the test window than the other ridges, and a second ridge being disposed between the first and third ridges.

19. The test apparatus of claim 14, wherein the first section includes multiple attachment projections that are configured to attach to corresponding attachment receptacles disposed on the second section, wherein dimensions of the attachment projections and attachment receptacles are configured such that in the attached configuration, the set of projections produces the pressure gradient that allows the sample fluid to flow at the predetermined flow rate along the length of the test strip.

20. The test apparatus of claim 14, wherein the inner face of the second section includes a second projection around the sample port, the second projection configured to inhibit a flow of components of the sample of whole blood along the length or width of the test strip.