USES OF WATER-DISPERSIBLE SILICA NANOPARTICLES FOR ATTACHING BIOMOLECULES

• US 20100209946A1
• Filed: 04/17/2008
• Published: 08/19/2010
• Est. Priority Date: 04/19/2007
• Status: Abandoned Application

• Current Assignee: 3M Innovative Properties Company (3M Company)
• Original Assignee: 3M Innovative Properties Company (3M Company)
• Inventors: Legatt, Michelle L., Schultz, William J., Jing, Naiyong, Zhang, Yifan, Guo, Chunmei
• Application Number:
  US12/595,858
• Publication Number:
  US 20100209946A1
• Time in Patent Office: Days
• US Class Current: 435/7.330
• CPC Class Codes: B82Y 5/00 Nanobiotechnology or nanomedicine, e.g. protein engineering or drug delivery
• CPC Class Codes: G01N 33/54346 Nanoparticles
• CPC Class Codes: G01N 33/54353 with ligand attached to the carrier via a chemical coupling agent coatings G01N33/54393

Abstract

Uses of silica nanoparticles functionalized with water-dispersible groups, shielding groups, and biomolecule-binding groups.

Claims

1. A method of capturing a target analyte, the method comprising:

providing water-dispersible nanoparticles, each comprising a silica surface having functional groups attached to the surface through nonreversible covalent bonds, wherein the functional groups comprise;

biomolecule-binding groups for attaching a biomolecule;

water-dispersible groups in a sufficient amount to provide water dispersibility to the nanoparticles; and

shielding groups distinct from the water-dispersible groups, wherein the bound shielding groups do not include amide groups and/or urea groups;

contacting the water-dispersible nanoparticles with a biomolecule under conditions effective to covalently bond the biomolecule to one or more biomolecule-binding groups, wherein the biomolecule is a capture agent for a target analyte; and

contacting the water-dispersible nanoparticles having the biomolecule capture agent covalently bonded thereto with a sample suspected of containing a target analyte;

with the proviso that the biomolecule-binding groups do not include aliphatic amine and/or maleimide groups having less than 6 carbon atoms, which are capable of covalently bonding to a biomolecule when the water-dispersible and/or shielding groups include poly(alkylene oxide)-containing groups.

2. The method of claim 1 wherein the target analyte comprises a microbe.

3. The method of claim 2 wherein the microbe comprises a bacterium.

4. The method of claim 3 wherein the bacterium comprises Staphylococcus aureus.

5. The method of claim 1 wherein the shielding groups comprise poly(alkylene oxide)-containing groups, ethylene glycol ether-containing groups, poly(ethylene oxide) ether-containing groups, ethylene glycol lactate-containing groups, sugar-containing groups, polyol-containing groups, crown ether-containing groups, oligo glycidyl-containing groups, hydroxyl acrylamide-containing groups, organosulfonate-containing groups, organocarboxylate-containing groups, or combinations thereof.

6. The method of claim 5 wherein the shielding groups comprise poly(alkylene oxide)-containing groups.

7. The method of claim 6 wherein the poly(alkylene oxide)-containing shielding groups comprise poly(ethylene oxide)-containing groups.

8. The method of claim 1 wherein the water-dispersible groups comprise carboxylic acid groups, sulfonic acid groups, phosphonic acid groups, salts thereof, or combinations thereof.

9. The method of claim 1 wherein the biomolecule is bonded to one or more biomolecule-binding groups through nonreversible covalent bonding.

10. The method of claim 1 wherein the nanoparticles have a particle size of no greater than 200 nm.

11. The method of claim 1 wherein the biomolecule-binding groups comprise functional groups selected from amines, hydrazines, hydroxyl groups, sulfones, aldehydes, alcohols, oxyranes, halides, N-oxysuccinimides, acrylates, acrylamides, alpha,beta-ethylenically or acetylenically unsaturated groups with electron withdrawing groups, carboxylates, esters, anhydrides, carbonates, oxalates, aziridines, epoxy groups, N-substituted maleimides, azlatones, and combinations thereof.

12. The method of claim 11 wherein the biomolecule-binding groups comprise functional groups selected from vinyl sulfones, epoxy groups, acrylates, amines, or combinations thereof.

13. The method of claim 11 wherein the biomolecule-binding groups comprise alpha-beta ethylenically unsaturated groups and electron withdrawing groups.

14. The method of claim 13 wherein the electron withdrawing groups comprise carbonyls, ketones, esters, amides, —

SO₂—

, —

SO—

, —

CO—

CO—

, —

CO—

COOR, sulfonamides, halides, trifluoromethyl, sulfonamides, halides, maleimides, maleates, or combinations thereof.

15. The method of claim 14 wherein the biomolecule-binding groups are acrylates or alpha,beta-unsaturated ketones.

16. The method of claim 1 wherein the biomolecule-binding groups comprise a nontertiary aromatic amine group and/or an aromatic hydrazine group.

17. The method of claim 1 wherein the biomolecule is aldehyde functional and covalently bonds to the biomolecule-binding group to form —

Ar—

N═

C(H)-biomolecule, or —

Ar—

NH—

N═

C(H)-biomolecule wherein Ar is an aryl group.

18. The method of claim 1 wherein the biomolecule-binding groups having a biomolecule covalently bonded thereto comprise a biotin-containing group covalently bonded to the surface of the nanoparticle through amine-functionalized groups.

19. The method of claim 1 wherein the nanoparticles further comprise a reporter group attached to the silica surface.

20. The method of claim 19 wherein the reporter group comprises a fluorescent group.

21. The method of claim 1 wherein prior to contacting the water-dispersible nanoparticle with a biomolecule, the method comprises oxidizing the biomolecule to form an aldehyde-functional biomolecule.

22. The method of claim 21 wherein the biomolecule is an antibody.

23. The method of claim 1 wherein contacting the water-dispersible nanoparticle with a biomolecule comprises contacting the water-dispersible nanoparticles with a plurality of antibodies of different specificities.

24. A method of attaching a biomolecule to nanoparticles, the method comprising:

providing silica nanoparticles, each comprising a surface;

providing a water-dispersible compound comprising a water-dispersible group and a surface-bonding group;

providing a biomolecule-binding compound comprising a biomolecule-binding group and a surface-bonding group;

providing a shielding compound comprising a shielding group and a surface-bonding group, wherein the shielding compound is distinct from the water-dispersible compound;

covalently bonding a plurality of the biomolecule-binding groups, water-dispersible groups, and shielding groups to the surface of a plurality of the silica nanoparticles through nonreversible covalent bonds between the surface-bonding groups and the surface;

wherein the bound shielding groups do not include amide groups and/or urea groups; and

contacting the water-dispersible nanoparticles with a biomolecule under conditions effective to covalently bond the biomolecule to one or more biomolecule-binding groups;

with the proviso that the biomolecule-binding groups do not include aliphatic amine and/or maleimide groups having less than 6 carbon atoms, which are capable of covalently bonding to a biomolecule when the water-dispersible and/or shielding groups include poly(alkylene oxide)-containing groups.

25. The method of claim 24 wherein the biomolecule is bonded to one or more biomolecule-binding groups through nonreversible covalent bonding.

26. The method of claim 24 wherein the biomolecule is a capture agent for a target biological analyte.

27. The method of claim 26 wherein the biomolecule capture agent is an antibody.

28. The method of claim 24 wherein the biomolecule is a target biological analyte.

29. The method of claim 24 further comprising:

providing a reporter molecule comprising a reporter group and a surface-bonding group; and

covalently bonding a plurality of the reporter groups to the surface of a plurality of the silica nanoparticles through the surface-bonding groups.

30. The method of claim 29 wherein the reporter group comprises a fluorescent group.

31. The method of claim 30 wherein the shielding compound is covalently bonded to the surface of the solid support material prior to the reporter molecule being bonded thereto.

32. The method of claim 24 wherein the biomolecule-binding groups comprise functional groups selected from amines, hydrazines, hydroxyl groups, sulfones, aldehydes, alcohols, oxyranes, halides, N-oxysuccinimides, acrylates, acrylamides, alpha,beta-ethylenically or acetylenically unsaturated groups with electron withdrawing groups, carboxylates, esters, anhydrides, carbonates, oxalates, aziridines, epoxy groups, N-substituted maleimides, azlatones, and combinations thereof.

33. The method of claim 32 wherein the biomolecule-binding groups comprise functional groups selected from vinyl sulfones, epoxy groups, acrylates, amines, or combinations thereof.

34. The method of claim 32 wherein the biomolecule-binding groups comprise alpha-beta ethylenically unsaturated groups and electron withdrawing groups.

35. The method of claim 34 wherein the electron withdrawing groups comprise ketones, esters, amides, —

SO₂—

, —

SO—

, —

CO—

CO—

, —

CO—

COOR, sulfonamides, halides, trifluoromethyl, sulfonamides, halides, maleimides, maleates, or combinations thereof.

36. The method of claim 35 wherein the biomolecule-binding groups are acrylates or alpha,beta-unsaturated ketones.

37. The method of claim 32 wherein the biomolecule-binding groups comprise a nontertiary aromatic amine group and/or an aromatic hydrazine group.

38. The method of claim 37 wherein the biomolecule is aldehyde functional and covalently bonds to the biomolecule-binding group to form —

Ar—

N═

C(H)-biomolecule, or —

Ar—

NH—

N═

C(H)-biomolecule wherein Ar is an aryl group.