Microanalytical device containing a membrane for molecular identification

• US 6,607,644 B1
• Filed: 10/31/2000
• Issued: 08/19/2003
• Est. Priority Date: 10/31/2000
• Status: Active Grant

• Current Assignee: Agilent Technologies, Inc.
• Original Assignee: Agilent Technologies, Inc.
• Inventors: Apffel, James A. Jr.
• Primary Examiner(s): Nguyen, Nam
• Assistant Examiner(s): NOGUEROLA, ALEXANDER STEPHAN
• Application Number:
  US09/703,431
• Time in Patent Office: 1,022 Days
• Field of Search: 204/450, 204/451, 204/600, 204/601, 422/50, 422/100
• US Class Current: 204/451
• CPC Class Codes: G01N 33/521 Single-layer analytical elements

Abstract

The invention provides a microanalytical device for analyzing a fluid sample containing at least one analyte molecule. The device is constructed from a substrate and a cover plate each having a substantially planar surface and a microchannel formed therein. The cover plate is placed over the substrate such that the cover plate microchannel is arranged in opposing relationship with the substrate microchannel. A membrane is interposed between the substrate and the cover and has at least one pore sized to allow passage of the analyte molecule from the substrate microchannel to the cover plate microchannel. An analyte altering moiety is attached to an interior surface of the pore and is capable of chemically altering the analyte molecule. Also disclosed are a method for chemically altering and transporting an analyte molecule in a fluid and a method for identifying a plurality of biomolecules.

Claims

1. A microanalytical device for analyzing a fluid sample containing at least one analyte molecule, comprising:

a substrate having a substantially planar substrate surface and a substrate microchannel formed in the substrate surface, wherein the substrate microchannel extends from an upstream terminus to a downstream terminus;

a cover plate having a substantially planar cover plate surface and a cover plate microchannel formed on the cover plate surface, wherein the cover plate surface is arranged over the substrate surface;

an inlet port in fluid communication with the substrate microchannel;

a membrane interposed between the substrate and the cover plate and having at least one pore sized to allow passage of the analyte molecule from the substrate microchannel to the cover plate microchannel, wherein the at least one pore is located downstream from the inlet port; and

an analyte altering moiety attached to an interior surface of the at least one pore, wherein the moiety is capable of chemically altering the analyte molecule.

2. The microanalytical device of claim 1, wherein the substrate microchannel is substantially parallel to the cover plate microchannel.

3. The microanalytical device of claim 2, wherein the membrane has a plurality of pores each sized to allow passage of fluid sample from the substrate microchannel to the cover plate microchannel.

4. The microanalytical device of claim 1, wherein the analyte altering moiety is adapted to digest the analyte molecule to form at least two analyte components.

5. The microanalytical device of claim 4, wherein the cover plate microchannel is adapted to separate the at least two analyte components.

6. The microanalytical device of claim 4, wherein the analyte altering moiety is an enzyme.

7. The microanalytical device of claim 6, wherein the enzyme is a proteolytic enzyme.

8. The microanalytical device of claim 1, wherein each pore of the membrane has a diameter in the range of about 10 angstroms to about 500 angstroms.

9. The microanalytical device of claim 1, wherein the membrane comprises a support that permits the fluid sample to diffuse therethrough and at least one monomolecular layer carried on a surface of the support, the at least one monomolecular layer being at least a portion of an active control structure.

10. The microanalytical device of claim 9, wherein the active control structure is controlled electrophorectically, electroosmotically or by application of pressure.

11. The microanalytical device of claim 1, wherein the analyte molecule is a biomolecule.

12. The microanalytical device of claim 11, wherein the biomolecule is peptidic.

13. The microanalytical device of claim 11, wherein the analyte molecule is nucleotidic.

14. The microanalytical device of claim 1, wherein the analyte molecule is oliogomeric or polymeric.

15. The microanalytical device of claim 1, wherein the fluid sample contains a first analyte molecule and a second analyte molecule.

16. The microanalytical device of claim 15, wherein the substrate microchannel is adapted to separate the first and second analyte molecules.

17. The microanalytical device of claim 1, wherein the substrate microchannel is adapted to chemically alter the analyte molecule.

18. The microanalytical device of claim 17, wherein the substrate microchannel is adapted to digest the analyte molecule to form at least two analyte components.

19. The microanalytical device of claim 18, wherein the substrate is further adapted to separate the at least two analyte components.

20. The microanalytical device of claim 1, wherein the cover plate microchannel is adapted for fluid communication with a mass spectrometer to allow the fluid sample to be introduced thereto.

21. The microanalytical device of claim 1, wherein the substrate microchannel is adapted for fluid communication with a source of the fluid sample.

22. The microanalytical device of claim 1, wherein the substrate microchannel comprises a first-dimension microchannel and at least one second-dimension microchannel, each second-dimension microchannel fluidly communicating with the first-dimension microchannel.

23. The microanalytical device of claim 22, wherein the substrate microchannel comprises a plurality of second-dimension microchannels each fluidly communicating with the first-dimension microchannel at a different point.

24. The microanalytical device of claim 23, wherein all of the second-dimension microchannels are parallel to one another.

25. A method for chemically altering and transporting an analyte molecule in a fluid, comprising the steps of:

(a) introducing the molecule through an inlet port into a microchannel formed in a substantially planar substrate surface so that the molecule travels along the microchannel;

(b) transporting the molecule from the substrate microchannel in a non-parallel direction with respect to the substrate surface into a pore of a membrane, wherein the pore is located downstream from the inlet port;

(c) chemically altering the analyte molecule to form an altered molecule using a moiety attached to an interior surface of the pore; and

(d) collecting the altered molecule from the pore on a cover plate microchannel formed on a substantially planar cover plate surface of a cover plate, wherein the substantially planar cover plate surface is in opposing relation to the substantially planar substrate surface.

26. The method of claim 25, wherein step (c) comprises digesting the analyte molecule using the moiety attached to the interior surface of the pore.

27. The method of claim 26, wherein the attached moiety comprises an enzyme.

28. The method of claim 27, wherein the attached enzyme is a proteolytic enzyme.

29. The method of claim 25, wherein step (b) comprises electrophoresis and/or pressure.

30. A method for identifying a plurality of biomolecules, comprising the steps of:

(a) providing a substrate having a substantially planar substrate surface, the substrate having a substrate microchannel formed in the substrate surface;

(b) spatially resolving the biomolecules along the substrate microchannel;

(c) transporting the biomolecules from the substrate microchannel in a non-parallel direction with respect to the substrate surface into pores of a membrane while preserving the spatial resolution of the biomolecules;

(d) digesting the biomolecule in the pores of the membrane to form biomolecular components;

(e) collecting the biomolecular components from the pores while preserving the spatial resolution of the biomolecular components on a cover plate microchannel formed on a substantially planar cover plate surface of a cover plate, wherein the substantially cover plate surface is in opposing relation to the substantially planar substrate surface; and

(f) spatially resolving the biomolecular components in the cover plate microchannel.