Translucent solid matrix assay device for microarray analysis

• US 20040166494A1
• Filed: 02/24/2003
• Published: 08/26/2004
• Est. Priority Date: 02/24/2003
• Status: Active Grant

• Current Assignee: Gentel Biosciences Inc. (Intuitive Biosciences, Inc.)
• Original Assignee: PriTest Inc.
• Inventors: Green, Lawrence R.
• Granted Patent: US 6,861,251 B2
• 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 2219/00315 Microtiter plates
• CPC Class Codes: B01J 2219/00317 Microwell devices, i.e. having large numbers of wells
• CPC Class Codes: B01J 2219/00387 Applications using probes
• CPC Class Codes: B01J 2219/00527 Sheets
• CPC Class Codes: B01J 2219/00533 essentially rectangular
• CPC Class Codes: B01J 2219/00536 in the shape of disks
• CPC Class Codes: B01J 2219/00576 fluorophore
• CPC Class Codes: B01J 2219/00605 the compounds being directly bound or immobilised to solid supports
• CPC Class Codes: B01J 2219/0061 The surface being organic
• CPC Class Codes: B01J 2219/00612 the surface being inorganic
• CPC Class Codes: B01J 2219/00637 by coating it with another layer
• CPC Class Codes: B01J 2219/00659 Two-dimensional arrays
• CPC Class Codes: B01J 2219/00675 In-situ synthesis on the substrate
• CPC Class Codes: B01J 2219/00722 Nucleotides
• CPC Class Codes: B01J 2219/00725 Peptides
• CPC Class Codes: B01J 2219/0074 Biological products
• CPC Class Codes: B82Y 30/00 Nanotechnology for materials or surface science, e.g. nanocomposites
• CPC Class Codes: C40B 40/10 Libraries containing peptides or polypeptides, or derivatives thereof
• CPC Class Codes: C40B 50/14 Solid phase synthesis, i.e. wherein one or more library building blocks are bound to a solid support during library creation; Particular methods of cleavage from the solid support

Abstract

The present invention concerns methods and compositions relating to matrix arrays. In certain embodiments, the arrays are translucent. In other embodiments, the arrays are reconfigurable. In preferred embodiments, the arrays are translucent and reconfigurable. Reconfigurable arrays may be produced using small linker molecules, such as aptamers or affibodies, attached to the array substrate. Preferably, the small linker molecules bind to an IgG specific portion of antibodies. Such arrays may be used to detect any target that binds selectively or specifically to an IgG, allowing great flexibility of use. Translucent matrix arrays may utilize a translucent, colloidal form of nitrocellulose to coat the array substrate.

Claims

1. A method of making translucent matrix arrays comprising:

a) coating a substrate with a translucent layer of nitrocellulose; and

b) attaching one or more first probes to the nitrocellulose.

2. The method of claim 1, wherein the nitrocellulose is colloidal nitrocellulose.

3. The method of claim 1, wherein the substrate is glass.

4. The method of claim 3, further comprising sealing the translucent layer of nitrocellulose to the glass substrate.

5. The method of claim 1, wherein the substrate is plastic.

6. The method of claim 1, wherein the first probes are selected from the group consisting of antibodies, antibody fragments, FAb fragments, humanized antibodies, single-chain antibodies, chimeric antibodies, oligonucleotides, polynucleotides, nucleic acids, aptamers and affibodies.

7. The method of claim 1, wherein the nitrocellulose is dissolved in amyl acetate to coat the substrate.

8. The method of claim 1, wherein the matrix array is reconfigurable.

9. The method of claim 8, wherein the first probes are aptamers or affibodies.

10. The method of claim 9, wherein the first probes bind to IgG (immunoglobulin G).

11. The method of claim 1, further comprising exposing the first probes to one or more samples suspected of containing targets.

12. The method of claim 11, further comprising allowing the first probes to bind to the targets.

13. The method of claim 12, further comprising binding tagged second probes to the targets.

14. The method of claim 13, further comprising detecting signals from the tagged second probes.

15. A translucent matrix array made by a method comprising:

a) coating a substrate with a translucent layer of nitrocellulose; and

b) attaching one or more first probes to the nitrocellulose.

16. The translucent matrix array of claim 15, wherein the first probes are attached to the array in spots of about 300 μ

m in size.

17. The translucent matrix array of claim 16, wherein the probes attached to each spot are specific for a different target.

18. The translucent matrix array of claim 15, wherein the first probes are selected from the group consisting of antibodies, antibody fragments, FAb fragments, humanized antibodies, single-chain antibodies, chimeric antibodies, oligonucleotides, polynucleotides, nucleic acids, aptamers and affibodies.

19. A method of identifying targets using reconfigurable matrix arrays comprising:

a) attaching small linker molecules to a matrix array surface;

b) allowing one or more targets to bind to one or more first and second probes in an aqueous solution;

c) allowing said first probes to bind to the small linker molecules; and

d) detecting the presence of tagged second probes attached to the matrix array surface.

20. The method of claim 19, wherein all the first and second probes are selected to bind to the same target in a single assay.

21. The method of claim 19, wherein the first and second probes are selected to bind to two or more different targets in a single assay.

22. The method of claim 21, wherein the second probes that bind to different targets are distinguishably labeled.

23. The method of claim 19, wherein the small linker molecules are aptamers or affibodies.

24. The method of claim 23, wherein the small linker molecules bind to IgG.

25. The method of claim 24, wherein the reconfigurable matrix array may be used to detect any target that binds to an IgG.

26. The method of claim 25, further comprising:

(i) using said array to detect a first target;

(ii) washing said array to remove said first target; and

(iii) using said array to detect a second target.

27. The method of claim 26, further comprising using said array to detect a multiplicity of targets.

28. The method of claim 27, wherein said targets are detected sequentially.