Detecting biochemical reactions

• US 20030068639A1
• Filed: 08/30/2002
• Published: 04/10/2003
• Est. Priority Date: 08/31/2001
• Status: Abandoned Application

• Current Assignee: Infineon Technologies AG
• Original Assignee: Infineon Technologies AG
• Inventors: Ehben, Thomas, Martin, Alfred, Fuchs, Karin, Hanke, Hans Christian, Haneder, Thomas, Lehmann, Volker
• Application Number:
  US10/233,001
• Publication Number:
  US 20030068639A1
• Time in Patent Office: Days
• US Class Current: 435/6
• CPC Class Codes: B01J 19/0046 Sequential or parallel reactions, e.g. for the synthesis of polypeptides or polynucleotides; Apparatus and devices for combinatorial chemistry or for making molecular arrays synthesis methods per se C40B50/00
• CPC Class Codes: B01J 2219/00286 Reactor vessels with top and bottom openings
• CPC Class Codes: B01J 2219/00389 Feeding through valves
• CPC Class Codes: B01J 2219/00511 Walls of reactor vessels
• CPC Class Codes: B01J 2219/00576 fluorophore
• CPC Class Codes: B01J 2219/00578 electrophoric
• CPC Class Codes: B01J 2219/00585 Parallel processes
• CPC Class Codes: B01J 2219/00596 Solid-phase processes
• CPC Class Codes: B01J 2219/00605 the compounds being directly bound or immobilised to solid supports
• CPC Class Codes: B01J 2219/00612 the surface being inorganic
• CPC Class Codes: B01J 2219/00641 the porous medium being continuous, e.g. porous oxide substrates
• CPC Class Codes: B01J 2219/00659 Two-dimensional arrays
• CPC Class Codes: B01J 2219/00677 Ex-situ synthesis followed by deposition on the substrate
• CPC Class Codes: B01J 2219/00722 Nucleotides
• CPC Class Codes: B01J 2219/00725 Peptides
• CPC Class Codes: B01J 2219/00729 Peptide nucleic acids [PNA]
• CPC Class Codes: B01J 2219/00731 Saccharides
• CPC Class Codes: B01J 2219/0074 Biological products
• CPC Class Codes: B01L 2300/069 Absorbents; Gels to retain a fluid
• CPC Class Codes: B01L 2300/0829 Multi-well plates; Microtitration plates
• CPC Class Codes: B01L 3/50857 : using arrays or bundles of open capillaries for holding samples
• CPC Class Codes: B82Y 30/00 : Nanotechnology for materials or surface science, e.g. nanocomposites
• CPC Class Codes: C40B 40/06 : Libraries containing nucleotides or polynucleotides, or derivatives thereof
• CPC Class Codes: C40B 40/10 : Libraries containing peptides or polypeptides, or derivatives thereof
• CPC Class Codes: C40B 40/12 : Libraries containing saccharides or polysaccharides, or derivatives thereof
• CPC Class Codes: C40B 60/14 : for creating libraries
• CPC Class Codes: G01N 33/54346 : Nanoparticles
• CPC Class Codes: G01N 33/54373 : involving physiochemical end-point determination, e.g. wave-guides, FETS, gratings

Abstract

A method for detecting a biochemical reaction includes immobilizing a capture molecule on an inner wall of a pore selected from a multiplicity of pores extending between first and second opposed surfaces of a macroporous substrate and contacting an analyte with the capture molecule. A light is then directed into the pore. A change in a light transmission property of the pore is then detected. This change indicates a binding reaction between the analyte and the capture molecule.

Claims

1. A method for detecting a biochemical reaction, the method comprising:

immobilizing a capture molecule on an inner wall of a pore selected from a multiplicity of pores extending between first and second opposed surfaces of a macroporous substrate;

contacting an analyte with the capture molecule;

directing light into the pore; and

measuring a light-transmission property of the pore, the light-transmission property being indicative of a binding reaction between the analyte and the capture molecule.

2. The method of claim 1, further comprising selecting a pore diameter of the pore to be between 500 nanometers and 100 micrometers.

3. The method of claim 1, further comprising selecting the macroporous substrate to be a macroporous silicon substrate.

4. The method of claim 1, further comprising selecting the macroporous substrate to have a thickness between 100 micrometers and 5000 micrometers.

5. The method of claim 1, further comprising distributing the multiplicity of pores so as to have a pore density in a range between 10⁵ pores per square centimeter and 10⁸ pores per square centimeter.

6. The method of claim 1, further comprising selecting the pore to have an inner surface area in a range between 10 square micrometers and 30,000 square micrometers.

7. The method of claim 1, further comprising distributing the multiplicity of pores in a grid.

8. The method of claim 7, further comprising scanning the grid with externally controlled microvalves arranged to correspond to the grid.

9. The method of claim 1, further comprising:

providing a support plate in optical communication with the second surface;

providing a recording device on the support plate; and

providing a microprocessor in communication with the recording device, the microprocessor being configured for analysis of data collected by the recording device.

10. The method of claim 9, wherein providing a recording device comprises providing a CCD array in optical communication with the second surface.

11. The method of claim 10, further comprising tilting the CCD array relative to the second surface.

12. The method of claim 1, wherein directing light into the pore comprises providing a waveguide for coupling light directly into the pore.

13. The method of claim 12, wherein providing a waveguide comprises selecting a waveguide that covers a multiplicity of pores.

14. The method of claim 1, wherein measuring a light-transmission property comprises detecting light that has been reflected back into the pore.

15. The method of claim 1, wherein directing light into the pore comprises providing a planar light waveguide in optical communication with the pore.

16. The method of claim 1, wherein directing light into the pore comprises providing a laser diode in optical communication with the pore.

17. The method of claim 1, wherein directing light into the pore comprises providing a glass fiber in optical communication with the pore.

18. The method of claim 17, wherein providing a glass fiber comprises providing a fiber having a beveled end in optical communication with the pore.

19. The method of claim 18, wherein providing a glass fiber having a beveled end comprises selecting the bevel angle to be in the range between 35 degrees and 55 degrees relative to the first surface.

20. The method of claim 19, wherein selecting the bevel angle comprises selecting the angle to be approximately 45 degrees.

21. The method of claim 1, further comprising selecting the capture molecule from the group consisting of DNA, proteins, and ligands.

22. The method of claim 21, further comprising selecting the capture molecule to be an oligonucleotide probe.

23. The method of claim 22, further comprising:

derivatizing the substrate with epoxysilane, and

binding the oligonucleotide probe with a terminal group selected from the group consisting of an amino group and a thiol group.

24. The method of claim 1, further comprising selecting the analyte from the group consisting of DNA, RNA, PNA, saccharides, peptides, proteins, cell components, individual cells, multicellular organisms, and cell assemblages.

28. The apparatus of claim 22, further comprising automatic application and sampling devices for automatically scanning the pores.

29. The apparatus of claim 28, wherein the automatic application and sampling devices comprise externally controllable microvalves disposed on a grid.

25. An apparatus for detecting a biochemical reaction, the apparatus comprising:

a macroporous substrate having

a first surface,

a second surface opposed to the first surface, and

a multiplicity of pores extending between the first and second surfaces;

a light supply device for supplying light to a pore selected from the multiplicity of pores;

a capture molecule immobilized at an inner wall of the pore; and

a measuring device for measuring a light-transmission property of the pore, the light-transmission property being indicative of an occurrence of a binding reaction between an analyte and the capture molecule.

26. The apparatus of claim 25, wherein the macroporous substrate comprises a macroporous silicon substrate.

27. The apparatus of claim 25, wherein the pores are disposed in a grid.

30. The apparatus of claim 25, wherein the measuring device comprises a CCD array in optical communication with the second surface.

31. The apparatus of claim 30, wherein the CCD array is tilted relative to the second surface.

32. The apparatus of claim 25, wherein the light supply device comprises a waveguide disposed to couple light directly into the pore.

33. The apparatus of claim 25, wherein the light supply device comprises a waveguide arranged to couple light into a multiplicity of pores.

34. The apparatus of claim 32, wherein the waveguide comprises a planar waveguide disposed on the first surface.

35. The apparatus of claim 25, wherein the light supply device comprises a laser diode disposed to couple light into the pore.

36. The apparatus of claim 25, wherein the light supply device comprises a glass fiber having a beveled end, the beveled end being disposed to couple light into the pore.

37. The apparatus of claim 36, wherein the beveled end is beveled at an angle in the range between 35 degrees and 55 degrees.

38. The apparatus of claim 37, wherein the beveled end is beveled at an angle of approximately 45 degrees.

39. The apparatus of claim 25, wherein the capture molecule is selected from the group consisting of DNA, proteins, and ligands.

40. The apparatus of claim 39, wherein the capture molecule comprises an oligonucleotide probe.

41. The apparatus of claim 25,

further comprising a reflecting agent disposed in optical communication with the second surface, and

wherein the measuring device is disposed to detect light reflected back from the reflecting agent.