Enhanced isotachophoresis assays using additives with spatial gradients

Patent metadata

Field	Value
Publication number	US-10132775-B2
Application number	US-201514828487-A
Country	US
Kind code	B2
Filing date	Aug 17, 2015
Priority date	Oct 1, 2010
Publication date	Nov 20, 2018
Grant date	Nov 20, 2018

Abstract

Techniques for enhanced isotachophoresis assays using additives with spatial gradients include forming a concentration gradient of an additive along a channel from an input port to an output port. The channel is used for isotachophoresis with ions of a leading electrolyte having a first mobility greater than a mobility of an analyte, and ions of a trailing electrolyte having a second mobility less than the mobility of the analyte. The additive is different from both the leading electrolyte and the trailing electrolyte; and the additive has a third mobility that assures the analyte will encounter the additive. The method further comprises introducing a mixture of the trailing electrolyte and a sample including the analyte. The method further comprises applying an electric field to the channel; and, measuring the analyte.

First claim

What is claimed is:

1. An isotachophoresis method comprising:
(a) forming a concentration gradient of a matrix material along a channel, wherein the channel connects an input port to an output port, and wherein the input port is configured to receive a sample comprising an analyte;
(b) introducing ions into the channel, wherein the ions comprise
(i) a leading electrolyte having a first effective mobility magnitude greater than an effective mobility magnitude of the analyte, and
(ii) a trailing electrolyte having a second effective mobility magnitude less than the effective mobility magnitude of the analyte;

(c) contacting the analyte to the leading electrolyte;
(d) contacting the trailing electrolyte to the analyte; and
(e) applying an electric field to the channel,
wherein the matrix material affects at least one of the analyte, the leading electrolyte or the trailing electrolyte in at least a portion of the channel.

2. The method of claim 1 , wherein the matrix material comprises a stationary material.

3. The method of claim 2 , wherein the matrix material comprises a stationary polymeric material.

4. The method of claim 1 , wherein the matrix material is ion permeable.

5. The method of claim 1 , further comprising detecting the analyte.

6. The method of claim 1 , wherein the analyte is a protein.

7. The method of claim 1 , wherein the analyte is a nucleic acid.

8. The method of claim 1 , further comprising introducing an additional ion into the channel via an additional channel at a junction, wherein the additional ion is different from the leading electrolye, the trailing electrolye, and the analyte.

9. The method of claim 8 , wherein prior to the introducing the additional ion, the additional ion is substantially absent from the channel.

10. The method of claim 8 , wherein the additional ion comprises a weak base or a weak acid.

11. The method of claim 8 , wherein the introducing the additional ion is conducted after the analyte has migrated along the channel at least to the junction.

12. The method of claim 8 , wherein the introducing the additional ion is triggered by a feedback signal.

13. The method of claim 12 , wherein the feedback signal is triggered by detection of an optical signal or an electrical signal.

14. An apparatus comprising:
(a) a channel connecting an input port to an output port, wherein the input port is configured to receive a sample comprising an analyte;
(b) a concentration gradient of a matrix material located in the channel;
(c) a leading electrolyte, wherein the leading electrolyte has a first effective mobility magnitude greater than an effective mobility magnitude of the analyte;
(d) a trailing electrolyte, wherein the trailing electrolyte has a second effective mobility magnitude less than the effective mobility magnitude of the analyte; and
(e) an electric field generator capable of applying an electric field to the channel,
wherein the matrix material affects at least one of the analyte, the leading electrolyte or the trailing electrolyte in at least a portion of the channel.

15. The apparatus of claim 14 , wherein the matrix material comprises a stationary material.

16. The apparatus of claim 15 , wherein the matrix material comprises a stationary polymeric material.

17. The apparatus of claim 14 , wherein the matrix material is ion permeable.

18. The apparatus of claim 14 , wherein the matrix material is functionalized with at least one type of capture molecule.

19. The apparatus of claim 14 , wherein the apparatus further comprises a detector.

20. The apparatus of claim 14 , wherein the analyte is a protein.

21. The apparatus of claim 14 , wherein the analyte is a nucleic acid.

22. The apparatus of claim 14 , further comprising an additional channel connected to the channel.

23. The apparatus of claim 22 , further comprising an additional ion in the additional channel.

24. The apparatus of claim 22 , wherein the additional channel comprises a side channel connected to the channel at a junction and wherein the electric field generator is configured to apply an electric field to the side channel when the analyte migrates through the channel to the junction.

25. The apparatus of claim 23 , wherein the additional ion comprises a weak base or a weak acid.

26. The apparatus of claim 23 , wherein the additional ion is not present in the channel.

27. The apparatus of claim 24 , wherein the application of the electric field to the side channel is activated by a feedback signal.

28. The apparatus of claim 24 , wherein the feedback signal is triggered by detection of an optical signal or an electrical signal.

29. An isotachophoresis method comprising:
(a) forming a concentration gradient of a matrix material along a channel, wherein the channel connects an input port to an output port, and wherein the input port is configured to receive a sample comprising an analyte;
(b) introducing ions into the channel, wherein the ions comprise
(i) a leading electrolyte having a first effective mobility magnitude greater than an effective mobility magnitude of the analyte, and
(ii) a trailing electrolyte having a second effective mobility magnitude less than the effective mobility magnitude of the analyte;

(c) contacting the analyte to the leading electrolyte;
(d) contacting the trailing electrolyte to the analyte; and
(e) applying an electric field to the channel,
wherein the matrix material is functionalized with specific capture molecules.

30. The method of claim 29 , wherein the specific capture molecules are present in a concentration gradient along the channel.

31. The method of claim 29 , wherein the specific capture molecules comprise at least one of the group consisting of a macromolecule, a nucleic acid, an antibody, an antigen, biotin, avidin, a ligand, a receptor, a bait molecule, and a protein.

32. An isotachophoresis method comprising:
(a) forming a concentration gradient of a matrix material along a channel, wherein the channel connects an input port to an output port, and wherein the input port is configured to receive a sample comprising an analyte;
(b) introducing ions into the channel, wherein the ions comprise
(i) a leading electrolyte having a first effective mobility magnitude greater than an effective mobility magnitude of the analyte, and
(ii) a trailing electrolyte having a second effective mobility magnitude less than the effective mobility magnitude of the analyte;

(c) contacting the analyte to the leading electrolyte; and
(d) contacting the trailing electrolyte to the analyte; and (e) applying an electric field to the channel,
wherein the matrix material is functionalized with probe molecules.

Assignees

• Santiago Juan G
• Persat Alexandre
• Garcia Giancarlo
• Eid Charbel
• Univ Leland Stanford Junior

Inventors

• Santiago Juan G
• Persat Alexandre
• Garcia Giancarlo
• Eid Charbel

Classifications

• G01N27/4473

  by electric means · CPC title

• G01N27/44795 Primary

  Isoelectric focusing · CPC title

• G01N27/44756 Primary

  Apparatus specially adapted therefor · CPC title

• G01N27/44721

  by optical means · CPC title