Method and device for moving molecules by the application of a plurality of electrical fields

• US 5,750,015 A
• Filed: 03/13/1996
• Issued: 05/12/1998
• Est. Priority Date: 02/28/1990
• Status: Expired due to Term

• Current Assignee: Monogram Biosciences, Inc. (Laboratory Corporation Of America Holdings)
• Original Assignee: Soane Biosciences, Inc.
• Inventors: Soane, David S., Soane, Zoya M.
• Primary Examiner(s): Gorgos, Kathryn L.
• Assistant Examiner(s): STARSIAK, JOHN S
• Application Number:
  US08/615,642
• Time in Patent Office: 790 Days
• Field of Search: 204/547, 204/643, 204/454, 204/451, 204/601
• US Class Current: 204/454
• CPC Class Codes: B01D 57/02 by electrophoresis treatment of water, waste water, sewage or sludge by electrophoresis C02F1/469; electrophoretic production of compounds or non-metals C25B7/00; investigating or analysing materials by using electrophoresis G01N27/26
• CPC Class Codes: B01F 33/30 Micromixers
• CPC Class Codes: B01F 33/3031 using electro-hydrodynamic [EHD] or electro-kinetic [EKI] phenomena to mix or move the fluids
• CPC Class Codes: B01J 19/0093 Microreactors, e.g. miniaturised or microfabricated reactors laboratory containers with capillary fluid transport in microfabricated channels or chambers B01L3/5027
• CPC Class Codes: B01J 2219/00853 Employing electrode arrangements
• CPC Class Codes: B01J 2219/00912 by electrophoresis
• CPC Class Codes: B82Y 10/00 Nanotechnology for information processing, storage or transmission, e.g. quantum computing or single electron logic
• CPC Class Codes: B82Y 40/00 Manufacture or treatment of nanostructures
• CPC Class Codes: C07K 1/26 Electrophoresis
• CPC Class Codes: G01N 27/44752 Controlling the zeta potential, e.g. by wall coatings
• CPC Class Codes: G01N 27/44773 Multi-stage electrophoresis, e.g. two-dimensional electrophoresis
• CPC Class Codes: G03F 7/0002 Lithographic processes using patterning methods other than those involving the exposure to radiation, e.g. by stamping

Abstract

Devices and methods are disclosed for moving charged molecules through a medium by the application of a plurality of electrical fields of sufficient strength and applied for sufficient amounts of time so as to move the charged molecules through the medium. The devices although preferably small in size, preferably generate large numbers (100 or more) of electrical fields to a movement area which preferably contains a liquid buffered or gel medium. Mixtures of charged molecules are pulled through the gel by the force of the electrical fields. The fields are preferably activated simultaneously or sequentially one after another at various speeds to create complex force field distributions or moving field waves along the separation medium. Charged molecules capable of moving quickly through the gel will be moved along by the faster moving field waves and be separated from slower moving molecules. The fields can be activated by computer software and can be used to move molecules away from and toward each other to obtain rapid and complex chemical synthesis, sequencing or reaction protocols.

Claims

1. A device for moving charged particles through a medium employing an electric field, said device comprising;

an electrically non-conductive solid support having an upper surface;

a main trench in said solid support extending downward from said upper surface;

a plurality of branch trenches connected to said main trench for moving charged particles into and out of said main trench; and

a plurality of electrodes positioned to be in electrical contact with a medium when present in said trenches.

2. A device according to claim 1 wherein said solid support comprises an organic polymer.

3. A device according claim 1 wherein at least one of said branch trenches crosses said main trench.

4. In a method for moving charged particles through a medium in a movement area comprising a trench of capillary dimensions using an electrical field with spaced apart electrodes positioned to be in electrical contact with a medium when present in said trench to produce said field, the improvement comprising:

supporting said medium with a non-porous polymer substrate having a substantially uncharged surface.

5. A method according to claim 4, wherein said polymer is polymethylmethacrylate, polycarbonate, polyethylene terephthalate or polystyrene and said polymer is optionally supported on glass.

6. A method according to claim 5, wherein said charged particles are separated during said moving into a plurality of components.

7. A method according to claim 4, wherein said charged particles are separated during said moving into a plurality of components.

8. A method according to claim 4, wherein said medium is a polymer gel.

9. A method according to claim 4, wherein said polymer is organic.

10. A method according to claim 4, wherein said charged particles are ions, molecules, cells or viruses.

11. In a method for moving charged particles through a medium in a movement area comprising a trench of capillary dimensions using an electrical field with spaced apart electrodes positioned to be in electrical contact with a medium when present in said trench to produce said field, the improvement comprising:

supporting said medium with a polymethylmethacrylate card.

12. A method according to claim 11, wherein said capillary dimensions are an inner diameter of from 25 to 100μ

.

13. A method according to claim 11, wherein said electrical field is created by a plurality of electrodes at opposite ends of said trench and along said trench.

14. A device for moving charged particles through a medium employing an electrical field, said device comprising:

a polymer solid substrate having an upper surface, wherein said upper surface of said polymer solid substrate is substantially uncharged;

a main trench of capillary dimensions in said substrate having opposite ends;

a pair of electrodes positioned to be in electrical contact with a medium when present in said trench, with one electrode proximal to one end of said trench and the other electrode proximal to the other end of said trench;

means for connecting said electrodes to a source of electricity; and

means for introducing and removing liquid from said trench.

15. A device according to claim 14, wherein said polymer is organic.

16. A device according to claim 15 wherein said organic polymer is polymethylmethacrylate, polycarbonate, polyethylene terephtahalate or polystryrene and said organic polymer is optionally supported on glass.

17. A device according to claim 16, wherein said capillary dimensions are a diameter of from 25 to 100μ

.

18. A device according to claim 14, wherein said charged particles are ions, molecules, cells or viruses.

19. A device for moving charged particles through a medium employing an electrical field, said device comprising:

an organic polymer solid substrate having an upper surface, wherein said upper surface of said organic polymer is substantially uncharged;

a main trench in said substrate extending downward from said upper surface, having opposite ends, said trench having a depth of about 5 and 25μ and

extending across said substrate;

a pair of electrodes positioned to be in electrical contact with a medium when present in said trench, with one electrode proximal to one end of said trench and the other electrode proximal to the other end of said trench;

means for connecting said electrodes to a source of electricity; and

ports for liquid transfer proximal to each end of said trench for liquid transport or a reservoir at each end of said trench.

20. A device according to claim 19, wherein said organic polymer substrate is polymethylmethacrylate.

21. A device according to claim 19, wherein said trench includes a gel for gel electrophoresis.

22. A device according to claim 19, further comprising:

at least one lateral branch trench crossing said main trench; and

at least one additional pair of electrodes positioned to be in electrical contact with a medium when present in said lateral branch trench, each additional pair proximal to opposite ends of each of said lateral branch trenches; and

means for connecting each of said additional pairs of electrodes to a source of electricity.

23. A device according to claim 22, further comprising:

an electronic computer for controlling the electricity delivered to each of said electrodes connected to said electrode connecting means.

24. A device according to claim 22, wherein base of said at least one lateral branch trench comprises reactant material.

25. A device according to claim 24, wherein said charged reactant material is labeled.

26. A device for moving charged particles through a medium employing an electrical field, said device comprising:

a polymethylmethacrylate card having an upper surface, wherein said upper surface of said substrate is substantially uncharged;

a main trench in said substrate extending downward from said upper surface, having opposite ends, said trench having capillary dimensions and extending across said substrate;

a pair of electrodes positioned to be in electrical contact with a medium when present in said trench, with one electrode proximal to one end of said trench and the other electrode proximal to the other end of said trench;

at least one lateral branch trench crossing said main trench; and

at least one additional pair of electrodes positioned to be in electrical contact with a medium when present in said lateral branch trench, each additional pair proximal to opposite ends of each of said lateral branch trenches;

means for connecting said electrodes to a source of electricity; and

ports for liquid transfer proximal to each end of said trench and each said lateral branch for liquid transport or a reservoir proximal to each end of said trench and each said lateral branch.

27. A device according to claim 26, said device further comprising:

an electronic computer for controlling the electricity delivered to each of said electrodes connected to said electrode connecting means.

28. A device according to claim 26, wherein said main trench contains a gel electrophoresis medium.

29. A device according to claim 28, wherein said gel electrophoresis medium is polyacrylamide.

30. In a capillary electrophoresis device comprising a capillary and electrodes positioned to be in electrical contact with a medium when present in said capillary and proximal to opposite ends of said capillary, the improvement which comprises:

a capillary of polymethylmethacrylate.

31. A device for moving charged particles through a medium employing an electrical field, said device comprising:

an organic polymer solid substrate having an upper surface, wherein said upper surface of said organic polymer solid substrate is substantially uncharged;

a main trench in said substrate extending downward from said upper surface having opposite ends, said trench having capillary dimensions and extending across said substrate;

a pair of electrodes positioned to be in electrical contact with a medium when present in said main trench, with one electrode proximal to one end of said main trench and the other electrode proximal to the other end of said main trench;

at least one latenal branch trench crossing said main trench, wherein said at least one lateral branch trench comprises a charged reactant material;

at least one additional pair of electrodes positioned to be in electrical contact with a medium when present in said lateral branch trench, each additional pair proximal to opposite ends of each of said lateral branch trenches;

means for connecting said electrodes to a source of electricity; and

ports for liquid transfer proximal to each end of said trench for liquid transport or a reservoir at each end of said trench.

32. A method for moving charged particles through a medium in a controlled manner in a device comprising:

at least one branched movement area comprising a main trench and at least two branch trenches, wherein said medium is present in said movement area;

at least one electrode positioned in each of said trenches to be in electrical contact with said medium when present in said movement area to apply at least two electric fields in different directions to said medium in said movement area; and

means for introducing liquid into and removing liquid from said movement area;

said method comprising;

introducing sample comprising said charged particles into said medium present in said movement area; and

applying at least two different electric fields to said medium in a manner sufficient to move said particles through said medium in a controlled manner.

33. The method according to claim 32, wherein said medium is selected from the group consisting of buffer solution, polymeric solution, surfactant micellular dispersion and gel.

34. The method according to claim 32, wherein said at least one branch trench comprises a charged reactant material.

35. The method according to claim 34, wherein said charged reactant material is labeled.

36. A method for moving charged particles through a medium in a controlled manner in a device comprising:

at least one branched movement area comprising a central trench and at least one branch comprising charged reactant material at its base, wherein said medium is present in said movement area;

at least two pairs of electrodes positioned to be in electrical contact with said medium in said movement area to apply at least two electric fields in different directions to said medium in said movement area; and

means for introducing liquid into and removing liquid from said movement area;

said method comprising;

introducing sample comprising said charged particles into said medium present in said movement area;

applying a central trench electric field to said medium to move said sample to a site at least proximal to the intersection of said central trench and latent branch;

applying a lateral branch electric field to said medium to move said charged reactant material to said reaction site.

37. A method according to claim 36, wherein when said charged reactant material reaches said reaction site, a reaction occurs between said charged reactant material and said sample to produce a reaction product.

38. A method according to claim 37, wherein said method further comprises detecting said reaction product at a site in said central trench.

39. A method according to claim 37, wherein at least a portion of said reaction between said sample and charged reactant material occurs in the absence of an applied electric field.

40. A method according to claim 37, further comprising applying said central trench electric field in a manner sufficient to move said reaction product to a second reaction site at the intersection of said main trench and a second lateral branch.

41. A device for moving particles through a medium employing an electrical field, said device comprising:

a non-porous polymer substrate having an upper surface;

a main trench in said substrate extending downward from said upper source having opposite ends, said trench having capillary dimensions and extending across said substrate;

a pair of electrodes positioned to be in electrical contact with a medium when present in said main trench, with one electrode proximal to one end of said main trench and the other electrode proximal to the other end of said main trench;

at least two branch trenches connected to said main trench;

at least one electrode positioned in each of said branch trenches so as to be in electrical contact with a medium present in said branch trenches;

means for connecting said electrodes to a source of electricity; and

ports for liquid transfer proximal to each end of said trench for liquid transport or a reservoir at each end of said trench.

42. A device according to claim 41, said device further comprising:

an electronic computer for controlling the electricity delivered to each of said electrodes.

43. A device according to claim 41, wherein at least said main trench contains a gel electrophoresis medium.

44. A method for moving particles through a medium in a controlled manner in a device comprising:

at least one branched movement area comprising main trench and at least two branch trenches connected to said main trench, wherein said medium is present in said movement area;

electrodes positioned in said trenches to be in electrical contact with said medium in said movement area to apply at least two electric fields in different directions to said medium in said movement area; and

means for introducing liquid into and removing liquid from said movement area;

said method comprising;

introducing sample comprising said particles into said medium present in said movement area; and

applying at least two different electric fields to said medium in a manner sufficient to move said particles through said medium in a controlled manner.

45. The method according to claim 44, wherein said medium is selected from the group consisting of buffer solution, polymeric solution, surfactant micellular dispersion and gel.

46. The method according to claim 44, wherein said at least one branch trench comprises a reactant material.

47. The method according to claim 46, wherein said charged reactant material is labeled.