Blood cell barrier for a lateral flow device

• US 8,535,617 B2
• Filed: 11/30/2007
• Issued: 09/17/2013
• Est. Priority Date: 11/30/2007
• Status: Active Grant

• Current Assignee: Kimberly-Clark Worldwide Incorporated (Kimberly-Clark Corporation)
• Original Assignee: Kimberly-Clark Worldwide Incorporated (Kimberly-Clark Corporation)
• Inventors: MacDonald, J. Gavin, Smith, Molly K.
• Primary Examiner(s): Merkling, Sally
• Application Number:
  US11/948,127
• Publication Number:
  US 20090142229A1
• Time in Patent Office: 2,118 Days
• Field of Search: None
• US Class Current: 422/420
• CPC Class Codes: G01N 33/491 by separating the blood components G01N15/05 takes precedence

Abstract

A lateral flow device for analyzing a whole blood sample is provided. More specifically, the lateral flow device contains a porous membrane that defines a barrier zone for separating red blood cells from blood plasma (includes plasma in which clotting factors haven been removed), which may then flow to a subsequent detection zone for analysis. The barrier zone is formed from a blood cell barrier composition that includes an unsaturated aliphatic fatty acid or an ester thereof. Without intending to be limited by theory, the present inventors believe such unsaturated aliphatic fatty acid molecules undergo autoxidation in the presence of air and hemoglobin to release peroxides (e.g., hydrogen peroxide) via oxidative saturation of double bonds. In turn, the released peroxides are believed to induce the formation of echinocytes or crenated blood cells. The crenated red blood cells are distorted in shape and less flexible and malleable than normal red blood cells, making them less able to penetrate into the pores of the porous membrane of the lateral flow device. Consequently, the stiffer, less flexible cells cannot move easily into the porous and become trapped at the surface of the membrane, while the liquid components of the sample flow and penetrate through the membrane to the detection zone for analysis.

Claims

1. A lateral flow device for detecting the presence of an analyte in a whole blood sample, the device comprising a porous membrane, wherein the porous membrane further comprises:

a barrier zone for separating red blood cells from the whole blood sample, wherein the barrier zone is formed from a blood cell barrier composition that comprises an unsaturated aliphatic fatty acid or ester thereof having a carbon chain of at least C₈ and more than one carbon-carbon double bond, wherein the blood cell barrier composition prevents the penetration of red blood cells into the porous membrane; and

a detection zone for detecting the analyte, the detection zone being located downstream from the barrier zone and in fluid communication therewith, wherein the barrier zone and detection zone are located in the same plane, further wherein the barrier zone is in the form of a first line and the detection zone is in the form of a second line, wherein the first and second lines are spaced apart from each other by a section of the porous membrane.

2. The lateral flow device of claim 1, wherein the unsaturated aliphatic fatty acid or ester thereof has a carbon chain of at least C₁₅ and at least two carbon-carbon double bonds.

3. The lateral flow device of claim 1, wherein the blood cell barrier composition includes an omega-3 fatty acid, omega-6 fatty acid, an ester of an omega-3 fatty acid, an ester of an omega-6 fatty acid, or a mixture thereof.

4. The lateral flow device of claim 1, wherein the blood cell barrier composition includes linoleic acid, α

-linolenic acid, eicosadienoic acid, eicosatrienoic acid, arachidonic acid, eicosapentaenoic acid, docosahexaenoic acid, an ester thereof, or a mixture thereof.

5. The lateral flow device of claim 1, wherein the blood cell barrier composition includes linoleic acid or an ester thereof.

6. The lateral flow device of claim 1, further comprising detection probes that are capable of generating a detection signal within the detection zone.

7. The lateral flow device of claim 6, wherein a receptive material is immobilized within the detection zone that is capable of binding to the detection probes or conjugates thereof.

8. The lateral flow device of claim 7, wherein the detection probes are conjugated with a specific binding member that preferentially binds to the analyte.

19. The lateral flow device of claim 1, wherein the unsaturated fatty acid or ester thereof is capable of undergoing autoxidation in the presence of air and the whole blood sample to release a peroxide.

20. The lateral flow device of claim 19, wherein the released peroxide is capable of inducing the formation of crenated blood cells in the sample.

21. The lateral flow device of claim 20, wherein the crenated blood cells are trapped by the barrier zone before reaching the detection zone.

22. The lateral flow device of claim 1, wherein the device is an in vitro test device.

23. The lateral flow device of claim 1, wherein the first and second lines are disposed in a direction that is substantially perpendicular to the flow of the sample through the device.

9. A method for detecting the presence of an analyte in a whole blood sample, the method comprising:

providing a lateral flow device that comprises a porous membrane, wherein the porous membrane further comprises a barrier zone and a detection zone in fluid communication therewith, wherein the barrier zone is formed from a blood cell barrier composition that comprises an unsaturated aliphatic fatty acid or ester thereof having a carbon chain of at least C₈ and more than one carbon-carbon double bond, wherein the blood cell barrier composition prevents the penetration of red blood cells into the porous membrane, wherein the barrier zone and detection zone are located in the same plane, and, further wherein the barrier zone is in the form of a first line and the detection zone is in the form of a second line, wherein the first and second lines are spaced apart from each other by a section of the porous membrane;

contacting the barrier zone with the whole blood sample so that blood plasma, is separated from the sample and flows to the detection zone; and

detecting the presence of the analyte within the blood plasma at the detection zone.

10. The method of claim 9, wherein the volume of the whole blood sample is less than about 10 microliters.

11. The method of claim 9, wherein the unsaturated aliphatic acid or ester thereof has a carbon chain of at least C₁₅ and at least two carbon-carbon double bonds.

12. The method of claim 9, wherein the blood cell barrier composition includes an omega-3 fatty acid, omega-6 fatty acid, an ester of an omega-3 fatty acid, an ester of an omega-6 fatty acid, or a mixture thereof.

13. The method of claim 9, wherein the blood cell barrier composition includes linoleic acid, α

-linolenic acid, eicosadienoic acid, eicosatrienoic acid, arachidonic acid, eicosapentaenoic acid, docosahexaenoic acid, an ester thereof, or a mixture thereof.

14. The method of claim 9, wherein the blood cell barrier composition includes linoleic acid or an ester thereof.

15. The method of claim 9, wherein the lateral flow device further comprises detection probes that are capable of generating a detection signal within the detection zone.

16. The method of claim 15, wherein a receptive material is immobilized within the detection zone that is capable of binding to the detection probes or conjugates thereof.

17. The method of claim 15, wherein the detection probes are conjugated with a specific binding member that preferentially binds to the analyte.

18. The method of claim 9, further comprising supplying a diluent to the lateral flow device to facilitate flow of the blood plasma to the detection zone.