Electrophoresis Devices And Methods For Focusing Charged Analytes

• US 20080087546A1
• Filed: 05/19/2004
• Published: 04/17/2008
• Est. Priority Date: 05/19/2003
• Status: Active Grant

• Current Assignee: Washington State University
• Original Assignee: Protasis Corp.
• Inventors: Ivory, Cornelius F., Strand, David, Leatzow, Dan M.
• Granted Patent: US 8,142,630 B2
• Time in Patent Office: Days
• US Class Current: 204/547
• CPC Class Codes: G01N 27/44795 Isoelectric focusing

Abstract

Devices are provided for separating and focusing charged analytes, comprising a separation chamber and two or more electrodes, for example, an electrode array. A membrane separates the separation chamber and the electrodes. The separation chamber of the device is configured, that is, the separation chamber has a shaped geometry, which serves to induce a gradient in an electric field generated by the electrodes in the electrode chamber. Optionally, molecular sieve is included in the separation chamber that is operative to shift the location at which a stationary focused band of a charged analyte forms under a given set of focusing process parameters. Methods are provided for separating and focusing charged analytes comprising introducing a first fluid comprising at least one charged analyte into the separation chamber of a device as just described, applying an electric field gradient to the charged analyte to focus the charged analyte in the electric field gradient.

Claims

1. An electrophoretic device comprising:

a separation chamber having a fluid inlet port and a fluid outlet port, with a flow path from the fluid inlet port to the fluid outlet port defining a fluid flow direction through the separation chamber, and

electrodes positioned in the electrode chamber and operative when energized to generate an electric field gradient in the separation chamber,

wherein the separation chamber has a non-uniform configuration along at least a portion of the flow path.

4. The electrophoretic device of claim 1, wherein the separation chamber has a non-uniform width and a non-uniform height.

12. The electrophoretic device of claim 1, further comprising an electrode chamber separated from the separation chamber by a permeable membrane, the electrodes being positioned in the electrode chamber.

15. The electrophoretic device of claim 12, wherein the electrode chamber has a non-uniform cross-section flow channel.

22. The electrophoretic device of claim 12, wherein the electrode chamber is defined by one or more non-linear walls.

23. The electrophoretic device of claim 22, wherein the one or more non-linear walls of the electrode chamber are curved in the direction of flow along the flow path.

26. The electrophoretic device of claim 1, further comprising molecular sieve in the separation chamber operative to shift the location at which a stationary focused band of charged analyte forms under a given set of focusing process parameters.

29. The electrophoretic device of claim 26, wherein the molecular sieve comprises a gel.

35. The electrophoretic device of claim 26, wherein the molecular sieve comprises zeolites.

38. The electrophoresis device of claim 1, wherein the electrodes comprise an electrode array.

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39. (canceled)

40. A method for focusing a analyte comprising:

providing a device for focusing a charged analyte comprising;

a non-uniform separation chamber having a fluid inlet port and a fluid outlet port,

with a flow path from the fluid inlet port to the fluid outlet port defining a fluid flow direction through the separation chamber, and

electrodes separated from the separation chamber and operative when energized to generate an electric field gradient in the separation chamber;

introducing a fluid comprising at least one analyte into the separation chamber via the inlet port; and

energizing the electrodes to establish an electric field gradient in the separation chamber to focus at least a portion of the analyte at a location along the flow path.

41. The method of claim 40, wherein the electrodes comprise an electrode array.

42. The method of claim 40, wherein the electric field gradient is changed during the course of focusing the charged analyte.

43. The method of claim 40, wherein the charged analyte comprises an uncharged material sorbed into a charged carrier.

44. The method of claim 40, wherein the separation chamber contains molecular sieve operative to shift the location at which a stationary focused band of a charged analyte forms under a given set of focusing process parameters.

53. The method of claim 40, wherein the electric field gradient is dynamically controlled.

45-46. -46. (canceled)

48. (canceled)

50. (canceled)

54. (canceled)

55. A method for focusing a charged analyte comprising:

providing a device for focusing a charged analyte comprising;

a non-uniform separation chamber and an electrode chamber separated from the separation chamber by a membrane, with electrodes positioned proximate the electrode chamber and operative to generate an electric field in the electrode channel;

introducing a first liquid comprising a plurality of charged analytes into the separation chamber; and

applying an electric field gradient to the plurality of charged analytes in the first liquid to focus the plurality of charged analytes in the electric field gradient into stationary focused bands of charged analytes, each charged analyte forming a stationary focused band.