Reactive probe chip, composite substrate and method for fabrication of the same

• US 20010039072A1
• Filed: 03/30/2001
• Published: 11/08/2001
• Est. Priority Date: 03/30/2000
• Status: Active Grant

• Current Assignee: Ebara Corporation
• Original Assignee: Ebara Corporation
• Inventors: Nagasawa, Hiroshi, Fukunaga, Akira, Hirose, Masayoshi
• Granted Patent: US 6,897,021 B2
• Time in Patent Office: Days
• US Class Current: 438/106
• CPC Class Codes: B01J 2219/00317 Microwell devices, i.e. having large numbers of wells
• CPC Class Codes: B01J 2219/00497 Features relating to the solid phase supports
• CPC Class Codes: B01J 2219/00527 Sheets
• CPC Class Codes: B01J 2219/00585 Parallel processes
• CPC Class Codes: B01J 2219/0059 Sequential 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 2219/00608 DNA chips
• CPC Class Codes: B01J 2219/0061 The surface being organic
• CPC Class Codes: B01J 2219/00612 the surface being inorganic
• CPC Class Codes: B01J 2219/00617 by chemical means
• CPC Class Codes: B01J 2219/00621 by physical means, e.g. trenches, raised areas
• CPC Class Codes: B01J 2219/00626 Covalent
• CPC Class Codes: B01J 2219/0063 Other, e.g. van der Waals forces, hydrogen bonding
• CPC Class Codes: B01J 2219/00637 by coating it with another layer
• CPC Class Codes: B01J 2219/00641 the porous medium being continuous, e.g. porous oxide substrates
• CPC Class Codes: B01J 2219/00644 the porous medium being present in discrete locations, e.g. gel pads
• CPC Class Codes: B01J 2219/00659 Two-dimensional arrays
• CPC Class Codes: B01J 2219/00722 Nucleotides
• CPC Class Codes: C40B 40/06 Libraries containing nucleotides or polynucleotides, or derivatives thereof
• CPC Class Codes: C40B 60/14 : for creating libraries
• CPC Class Codes: G01N 33/54346 : Nanoparticles
• CPC Class Codes: G01N 33/545 : Synthetic resin
• CPC Class Codes: G01N 33/552 : Glass or silica

Abstract

A reaction probe chip which is prepared by loading a reactive probe on fine pieces of carrier such as particles, tile-like plates and then arraying and immobilizing the reactive probe-loaded carrier on a base material. The carrier fine pieces such as particles, tile-like plates and the like are porous or have a reactive surface, and the base material is preferably a thin inorganic plate or a thin organic plate is disclosed.

The inorganic base material is preferably a glass slide or silicon wafer, and the organic base material is preferably a polyester film or polyethylene film. In case the porous carrier pieces are used, the reactivity of the inner surfaces of the porous carrier pores should be maintained during array or immobilization process of the reactive probe-loaded carrier.

A composite substrate characterized in that on at least a section of the surface thereof, a plurality of porous regions are orderly arranged as compartments by non-porous regions, or a plurality of non-porous regions are orderly arranged as compartments by porous regions is also disclosed. The porous solid is preferably porous glass or porous ceramic, the porous glass is preferably split-phase porous glass, and the surface is preferably flattened by a process such as polishing.

Claims

1. A reactive probe chip comprises:

a particulate carrier or a tile-like carrier loaded with a reactive probe, and a substrate, wherein said particulate carrier or said tile-like carrier is set on said substrate.

3. A reactive probe chip according to

claim 1

or

claim 2

, wherein said particulate carrier is a porous carrier particle.

4. A reactive probe chip according to

claim 1

or

claim 2

, wherein said particulate carrier is of a material with a surface having bonding ability.

5. A reactive probe chip according to

claim 4

, wherein said particulate particle is selected from a porous glass, silica gel or ion-exchange resin.

6. A reactive probe chip according to

claim 1

or

claim 2

, wherein said substrate is an inorganic or organic substrate.

7. A reactive probe chip according to

claim 1

, wherein said reactive probe is selected from the group consisting of DNA, RNA and PNA (peptide nucleic acid) and fragments thereof, oligonucleotides of any desired base sequence, antigens, antibodies, epitopes, enzymes, proteins and the polypeptide chains.

9. A reactive probe chip according to

claim 1

, wherein said carrier is porous particles, said reactive probe is loaded on the inner surfaces of the porous particle pores, and the probe-loaded particles are arrayed and immobilized in at least one of a plurality of microcompartments provided on a base material, while maintaining the reactivity of the inner surfaces of the porous carrier particle pores.

10. A reactive probe chip according to

claim 3

, wherein the pore size of the porous carrier particles ranges from 10 nm to 1 μ

m, and the particle size ranges from 1 μ

m to 100 μ

m.

11. A reactive probe chip according to

claim 1

or

claim 2

, wherein said carrier is a square or hexagonal or circular plate and has a diameter or a size of from 50 μ

m to 5 mm on each side.

2. A reactive probe chip comprises:

a particulate carrier or a tile-like carrier loaded with a reactive substance which has an ability to bond a detection target, and

a substrate, wherein said particulate carrier or a tile-like carrier is set on said substrate.

8. A reactive probe chip according to

claim 2

, wherein said reactive substance is selected from the group consisting of DNA, RNA and PNA (peptide nucleic acid) and fragments thereof, oligonucleotides of any desired base sequence, antigens, antibodies, epitopes, enzymes, proteins and the polypeptide chains.

12. A method for fabrication of a reactive probe chip which comprises the steps of:

immobilizing a reactive probe on particulate or tile-like carriers, and

arraying and immobilizing each of the loaded carriers in separate compartments on the base material.

13. A method for fabrication of a reactive probe chip according to

claim 12

, wherein said reactive substance is selected from the group consisting of enzymes, antigens, DNA, RNA and PNA (peptide nucleic acid) and fragments, antibodies, epitopes, proteins and peptides.

14. A method for fabrication of a reactive probe chip according to

claim 12

, wherein said oligonucleotides or proteins are synthesized on the carriers, and each carrier is arrayed and immobilized in separate compartments on the base material.

15. A method for fabrication of a reactive probe chip according to

claim 12

, wherein said carrier particles are of a material with a surface having bonding ability, and are selected from such as porous glass, silica gel or ion-exchange resin.

16. A method for fabrication of a reactive probe chip according to

claim 12

, wherein said carriers are porous carrier particles, and said particles loaded with the reactive probe are arrayed and immobilized using one or more same or different particles in at least one of a plurality of microcompartments provided on a base material while maintaining the reactivity of the inner surfaces of the carrier particle pores.

17. A method for fabrication of a reactive probe chip according to

claim 16

, wherein the pore size of the porous carrier particles ranges from 10 nm to 1 μ

m, and the particle size ranges from 1 μ

m to 100 μ

m.

18. A method for fabrication of a reactive probe chip according to

claim 12

, wherein the tile-like carriers have a square, hexagonal or circular shape and have a size of from 50 μ

m to 5 mm on each side or on diameter, and they are attached to and immobilized on the base material.

19. A method for fabrication of a reactive substance chip which comprises the steps of:

immobilizing a reactive substance which has an ability to bond a detection target on particulate or tile-like carriers, and

arraying and immobilizing each of the loaded carriers in separate compartments on the base material.

20. A method for fabrication of a reactive substance chip according to

claim 19

, wherein said reactive substance is selected front the group consisting of enzymes, antigens, DNA, RNA and PNA (peptide nucleic acid) and fragments, antibodies, epitopes, proteins and peptides.

21. A method for fabrication of a reactive substance chip according to

claim 20

, wherein said oligonucleotides or proteins are synthesized on the carriers, and each carrier is arrayed and immobilized in separate compartments on the base material.

22. A method for fabrication of a reactive substance chip according to

claim 19

, wherein said particulate carriers are of a material with a surface having bonding ability, and are selected from such as porous glass, silica gel or ion-exchange resin.

23. A method for fabrication of a reactive substance chip according to

claim 19

, wherein said carriers are porous carrier particles, and said particles loaded with the reactive substance are arrayed and immobilized using one or more same or different particles in at least one of a plurality of microcompartments provided on a base material while maintaining the reactivity of the inner surfaces of the carrier particle pores.

24. A method for fabrication of a reactive substance chip according to

claim 19

, wherein the pore size of the porous carrier particles ranges from 10 nm to 1 μ

m, and the particle size ranges from 1 μ

m to 100 μ

m.

25. A method for fabrication of a reactive substance chip according to

claim 19

, wherein the tile-like carriers have a square, hexagonal or circular shape and have a size of from 50 μ

m to 5 mm on each side or on diameter, and they are attached to and immobilized on the base material.

26. A method for fabrication of porous carrier particles loaded with a reactive substance, which comprises synthsizing oligo-nucleotides or proteins on the porous carrier particles by use of a solid phase method.

27. A composite substrate which comprises, on at least a section of the surface thereof, a plurality of porous regions arrayed on and comparted by non-porous regions, or a plurality of non-porous regions arrayed on and comparted by porous regions.

28. A composite substrate according to

claim 27

, wherein said composite substrate comprising both porous regions and non-porous regions and having a surface flattened by a process such as polishing.

29. A method for fabrication of a composite substrate according to

claim 27

wherein the separately formed porous solid is placed on prescribed regions of a non-porous substrate.

30. A method for fabrication of a composite substrate according to

claim 27

, wherein a porous solid precursor material is placed on prescribed regions of a non-porous substrate and the pores in said porous solid precursors is produced on the base material.

31. A method for fabrication of a composite substrate according to

claim 27

, wherein a plurality of the porous regions is formed in a plurality of predetermined regions on a substrate whose total surface is a porous solid precursor.