Devices and methods for focusing analytes in an electric field gradient II

• US 20060124459A1
• Filed: 01/12/2004
• Published: 06/15/2006
• Est. Priority Date: 01/15/2003
• Status: Abandoned Application

• Current Assignee: Protasis Corp.
• Original Assignee: Protasis Corp.
• Inventors: Strand, David, Leatzow, Dan M.
• Application Number:
  US10/542,509
• Publication Number:
  US 20060124459A1
• Time in Patent Office: Days
• US Class Current: 204/450
• CPC Class Codes: C07K 1/26 Electrophoresis
• CPC Class Codes: G01N 27/44795 Isoelectric focusing

Abstract

Devices are provided for separating and focusing analytes, comprising a separation chamber and electrodes separated from the separation chamber by a membrane. The electrodes are operative to generate an electric field in the separation chamber. Molecular sieve in the separation chamber is operative to shift the location at which a stationary focused band of the 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 separation chamber to focus the charged analyte at a location in the separation chamber. Methods are provided for separating and focusing un-charged (including inadequately charged) analytes, comprising introducing a fluid comprising at least the uncharged analyte and lipids, micelles and/or vesicles into the separation chamber of a device as just described, and applying an electric field gradient to the separation chamber to focus the analyte (in association with the lipids, micelles and/or vesicles) at a location in the separation chamber.

Claims

18. An electrophoretic device for focusing a charged analyte comprising:

a separation chamber comprising an inlet for introducing a first fluid into the separation chamber and an outlet for exiting the first liquid from the separation chamber;

an electrode chamber comprising an electrode array and an inlet for introducing a second liquid into the electrode chamber and an outlet for exiting the second liquid from the electrode chamber;

permeable material separating the separation and electrode chambers; and

molecular sieve in the separation chamber operative to shift the location at which a stationary focused band of a charged analyte forms under a given set of focusing process parameters.

19. A device for focusing a charged analyte comprising:

a first block comprising a first trough having an inlet for introducing a first liquid to the first trough and an outlet for exiting the first liquid from the first trough;

a second block comprising a second trough having an inlet for introducing a second liquid to the second trough and an outlet for exiting the second liquid from the second trough, the second trough further comprising an electrode array positioned in the second trough

wherein the first trough and the second trough are substantially coincident and form a channel when the first block is sealed to the second block; and

permeable material intermediate the first and second blocks,

wherein the permeable material divides the channel formed when the first block is sealed to the second block into a first chamber and a second chamber, the second chamber including the electrode array, and wherein the first chamber comprises 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.

20. A device for focusing a charged analyte comprising:

a first block having a first trough having an inlet for introducing a first liquid to the first trough and an outlet for exiting the first liquid from the first trough;

a second block having a second trough having an inlet for introducing a second liquid to the second trough and an outlet for exiting the second liquid from the second trough, the second trough further comprising an electrode array comprising a plurality of electrodes arranged linearly along the length of the second trough,

wherein the first trough and the second trough are substantially coincident and form a channel when the first block is sealed to the second block;

permeable material intermediate the first and second blocks;

a voltage controller for controlling the voltage applied to each electrode of the electrode array,

wherein the permeable material divides the channel formed when the first block is sealed to the second block into a first chamber and a second chamber, the second chamber including the electrode array, such that the first chamber and second chamber are in liquid communication and the first chamber is in electrical communication with the electrode array when the chambers are filled with a conductive liquid, and

wherein the first chamber comprises 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.

21. A device for focusing and separating charged analytes comprising:

a separation chamber comprising an inlet for introducing a first liquid into the separation chamber and an outlet for exiting the first liquid from the separation chamber;

an electrode chamber comprising an electrode array and an inlet for introducing a second liquid into the electrode chamber and an outlet for exiting the second liquid from the electrode chamber;

permeable material separating the separation and electrode chambers; and

molecular sieve in the separation chamber operative to separate bands of charged analyte of similar electrophoretic mobilities and different molecular weights.

22. A device for focusing a charged analyte comprising:

a separation chamber comprising an inlet for introducing a first fluid into the separation chamber and an outlet for exiting the first liquid from the separation chamber;

an electrode array isolated from the separation chamber and operative to establish an electric field gradient in the separation chamber; and

molecular sieve in the separation chamber operative to shift the location at which a stationary focused band of a charged analyte forms under a given set of focusing process parameters.

23. An electrophoresis device for focusing a charged analyte comprising:

a separation chamber;

means for generating an electric field in the separation chamber; and

molecular sieve in the first chamber operative to shift the location at which a stationary focused band of a charged analyte forms under a given set of focusing process parameters.

24-1. The method of claim 24, wherein the analyte is a charged analyte.

24-2. The method of claim 24, wherein the analyte is focused withy the aid of lipids, micelles or vesicles in the sample fluid.

46. A method for focusing a charged analyte comprising:

providing a device for focusing a charged analyte comprising;

a separation chamber comprising an inlet for introducing a first liquid into the separation chamber and an outlet for exiting the first liquid from the separation chamber;

an electrode chamber comprising an electrode array and an inlet for introducing a second liquid into the electrode chamber and an outlet for exiting the second liquid from the electrode chamber; and

permeable material separating the separation and electrode chambers;

introducing a first liquid comprising at least one charged analyte into the separation chamber; and

applying an electric field gradient to the charged analyte in the first liquid to focus the charged analyte in the electric field gradient,

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.

47. A method for focusing a charged analyte in a fluid medium comprising:

providing a device for focusing a charged analyte comprising;

a first block comprising a first trough having an inlet for introducing the first liquid to the first trough and an outlet for exiting the first liquid from the first trough;

a second block comprising a second trough having an inlet for introducing a second liquid to the second trough and an outlet for exiting the second liquid from the second trough, the second trough further comprising an electrode array positioned in the second trough,

wherein the first trough and the second trough are substantially coincident- and form a channel when the first block is sealed to the second block; and

permeable material intermediate the first and second blocks,

wherein the permeable material divides the channel formed when the first block is sealed to the second block into a first chamber and a second chamber, the second chamber including the electrode array;

introducing a first liquid comprising at least one charged analyte into the first chamber;

applying an electric field gradient to the charged analyte to cause the charged analyte to focus in a region of the first chamber,

wherein the first chamber comprises molecular sieve operative to shift the location at which a stationary focused band of charged analyte forms under a given set of focusing process parameters.

48. A method of separating charged analytes comprising:

providing a device for focusing a charged analyte comprising a separation chamber and electrodes separated from the separation chamber by a conductive membrane and operative to generate an electric field in the separation chamber;

introducing a first liquid comprising at least one charged analyte into the first 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 bands of charged analytes, each charged analyte forming a stationary focused band,

wherein the first chamber contains molecular sieve operative to shift the location at which each stationary focused band of charged analyte forms under a given set of focusing process parameters.

49. The method of claim 48, wherein stationary focused bands of charged analyte are separated on the basis of their molecular weights.

50. The method of claim 48, wherein the plurality of charged analytes comprises charged analytes that have the same charge to mass ratio.

51. The method of claim 48, wherein the plurality of charged analytes comprises charged analytes that have the same electrophoretic mobility.

52. The method of claim 48, wherein each stationary focused band of charged analyte is focused each at a stable location separate from the other charged analytes.

53. A method of separating charged analytes comprising:

providing a device for focusing a charged analyte comprising;

a separation chamber comprising an inlet for introducing a first liquid into the separation chamber and an outlet for exiting the first liquid from the separation chamber;

an electrode chamber comprising an electrode array and an inlet for introducing a second liquid into the electrode chamber and an outlet for exiting the second liquid from the electrode chamber; and

permeable material separating the separation and electrode chambers;

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 bands of charged analytes, each charged analyte forming a stationary focused band,

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

54. The method of claim 53, wherein the molecular sieve is operable to separate the stationary focused bands of charged analyte of similar electrophoretic mobilities and different molecular weights, each at a stable location separate from the other charged analytes.