MULTICOMPONENT PROTEIN MICROARRAYS

• US 20090088329A1
• Filed: 06/12/2008
• Published: 04/02/2009
• Est. Priority Date: 11/01/2002
• Status: Abandoned Application

• Current Assignee: Mcmaster University
• Original Assignee: Mcmaster University
• Inventors: Rupcich, Nicholas, Brennan, John D.
• Application Number:
  US12/138,193
• Publication Number:
  US 20090088329A1
• Time in Patent Office: Days
• US Class Current: 506/7
• 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 2219/00387 Applications using probes
• CPC Class Codes: B01J 2219/00527 Sheets
• CPC Class Codes: B01J 2219/00596 Solid-phase processes
• 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 2219/00725 Peptides
• CPC Class Codes: C40B 40/10 Libraries containing peptides or polypeptides, or derivatives thereof
• CPC Class Codes: C40B 60/14 for creating libraries
• CPC Class Codes: G01N 33/5436 with ligand physically entrapped within the solid phase liposomes G01N33/5432; immunological test elements G01N33/54386

Abstract

The present invention involves a multicomponent protein microarray comprising two or more components of a protein-based system entrapped within spots of a biomolecule compatible matrix arranged on a surface. Also included are methods of using the microarray for multicomponent analysis along with kits and machinery comprising the microarray.

Claims

1. A microarray comprising one or more spots of a biomolecule-compatible matrix having two or more components of a protein-based system entrapped therein, wherein the one or more spots are adhered to a surface.

2. The microarray according to claim 1, wherein the biomolecule-compatible matrix is a sol-gel.

3. The microarray according to claim 2, wherein the sol-gel is prepared from one or more organic polyol silanes.

4. The microarray according to claim 3, wherein the organic polyol silane is derived from one or more of sugar alcohols, sugar acids, saccharides, oligosaccharides and polysaccharides.

5. The microarray according to claim 3, wherein the organic polyol silane is derived from one or more of allose, altrose, glucose, mannose, gulose, idose, galactose, talose, ribose, arabinose, xylose, lyxose, threose, erythrose, glyceraldehydes, sorbose, fructose, dextrose, levulose, sorbitol, sucrose, maltose, cellobiose, lactose, dextran, (500-50,000 MW), amylose, pectin, glycerol, sorbitol, and trehelose.

6. The microarray according to claim 5, wherein the organic polyol silane is derived from one or more of glycerol, sorbitol, maltose and dextran.

7. The microarray according to claim 3, wherein the organic polyol silane is selected from one or more of diglycerylsilane (DGS), monosorbitylsilane (MSS), monomaltosylsilane (MMS), dimaltosylsilane (DMS) and a dextran-based silane (DS).

8. The microarray according to claim 7, wherein the organic polyol silane is selected from one or more of DGS and MSS.

9. The microarray according to claim 2, wherein the sol-gel is prepared from one or more of functionalized or non-functionalized alkoxysilanes;

functionalized or non-functionalized bis-silanes of the structure (RO)₃Si—

R′

—

Si(OR)₃, where R may be ethoxy, methoxy or other alkoxy groups and R′

is a functional group containing at least one carbon;

functionalized or non-functionalized chlorosilanes;

silicates; and

sugar, polymer, polyol or amino acid substituted silicates.

10. The microarray according to claim 9, wherein the sol gel is prepared from sodium silicate.

11. The microarray according to claim 1, wherein the matrix further comprises an effective amount of one or more additives.

12. The microarray according to claim 11, wherein the one or more additives are selected from one or more of humectants and protein stabilizing agents.

13. The microarray according to claim 12, wherein the one or more additives are selected from one or more organic polyols, hydrophilic, hydrophobic, neutral or charged organic polymers, block or randon co-polymers, polyelectrolytes, sugars and amino acids.

14. The microarray according to claim 13, wherein the one or more additives are selected from one or more of glycerol, sorbitol, sarcosine and polyethylene glycol.

15. The microarray according to claim 14, where the additive is glycerol.

16. The microarray according to claim 1, wherein the surface is a solid support made of glass, plastic, polymers, metals, ceramics, alloys or composites.

17. The microarray according to claim 16, wherein the surface is a solid support made of glass.

18. The microarray according to claim 17, wherein the glass is cleaned to substantially remove any organic matter and adsorbed metal ions.

19. The microarray according to claim 17, wherein the glass is modified with aminopropyltrithoxysilane (APTES), glycidoxyaminopropyltrimethoxysilane (GPS) or another suitable coupling agent that promotes adhesion of the microspots to the planar surface.

20. The microarray according to claim 19, wherein the glass is modified with glycidoxyaminopropyltrimethoxysilane (GPS).

21. The microarray according to claim 1, wherein the spots are spatially defined.

27. A kit, biosensor, micromachined device or medical device comprising the microarray according to claim 1.

28. A kit comprising one or more microarrays according to claim 1 and optionally, one or more of:

(a) reagents for use with the one or more microarrays;

(b) signal detection array-processing instruments;

(c) databases; and

(d) analysis and database management software;

29. A method of conducting a target discovery business comprising:

(a) providing one or more assay systems for identifying test substances by their ability to effect one or more protein based systems, said assay systems using one or more microarrays according to any one of claims 1-21;

(b) (optionally) conducting therapeutic profiling of the test substances identified in step (a) for efficacy and toxicity in animals; and

(c) licensing, to a third party, the rights for further drug development and/or sales or test substances identified in step (a), or analogs thereof.

22. A method of preparing a microarray comprising:

(a) combining two or more components of a protein-based system with one or more biomolecule-compatible precursor solutions; and

(b) applying the combination of (a) to a surface in a microarray format.

23. The method according to claim 22, further comprising:

(c) allowing the combination of (a) to gel on the surface.

24. The method according to claim 23, wherein the two or more components of a protein-based system and one or more biomolecule-compatible precursor solutions are combined with an effective amount of one or more additives.

25. A method of performing multi-component assays comprising:

(a) obtaining a biomolecule compatible microarray comprising a matrix having two or more components of a protein-based system entrapped therein;

(b) exposing the biomolecule-compatible microarray to one or more test substances; and

(c) detecting a change in the protein-based system.

26. The method according to claim 25, further comprising comparing the change in the protein based system to a control, wherein a change in the protein based system upon exposure to a reagent of interest compared to the control is indicative of the effect of the test substance on the protein based system.