Inverse emulsion methods of making polymeric imprint beads

• US 20030165882A1
• Filed: 04/11/2002
• Published: 09/04/2003
• Est. Priority Date: 04/11/2001
• Status: Active Grant

• Current Assignee: Aspira Biosystems, Inc. (Nanomune Technologies, Inc.)
• Original Assignee: Aspira Biosystems, Inc. (Nanomune Technologies, Inc.)
• Inventors: Huang, Chin-Shiou, Lynch, Casey C., Strikovsky, Alexander
• Granted Patent: US 7,105,289 B2
• Time in Patent Office: Days
• US Class Current: 435/6
• CPC Class Codes: G01N 33/54313 the carrier being characterised by its particulate form
• CPC Class Codes: G01N 33/54393 Improving reaction conditions or stability, e.g. by coating or irradiation of surface, by reduction of non-specific binding, by promotion of specific binding
• CPC Class Codes: G01N 33/545 Synthetic resin

Abstract

The present invention provides imprint bead compositions useful for capturing, isolating, detecting, analyzing and/or quantifying molecules in a sample. The imprint bead compositions comprise a matrix material having imprint cavities of a template molecule or molecules imprinted thereon wherein a substantial number of the imprint cavities are located at or near the surface of the matrix material.

Claims

1. A method of making an imprint bead comprising the steps of:

(a) emulsifying a composition comprising a polar solvent, a non-polar solvent, an organic monomer which is polymerizable in aqueous solution and a conjugate molecule, said conjugate molecule comprising a template moiety and a tail moiety, to form an inverse emulsion, wherein the template moiety is capable of partitioning into the noncontinuous phase of the inverse emulsion and the tail moiety is capable of partitioning into the continuous phase of the inverse emulsion;

(b) polymerizing the organic monomer; and

(c) removing the conjugate molecule.

2. The method of claim 1 wherein the polar solvent is water.

3. The method of claim 1 wherein the composition further comprises a surfactant.

4. The method of claim 3 wherein the surfactant is a nonionic surfactant.

5. The method of claim 4 wherein the nonionic surfactant is selected from the group consisting of sorbitan oleates, block copolymers, glycerol esters, Arlacel 83, Span 80, Hypermer B239 and Hypermer B246SF.

6. The method of claim 1 wherein the composition further comprises a dispersant.

8. The method of claim 1 wherein the organic monomer is selected from the group consisting of styrene, methyl methacrylate, 2-hydroxyethyl methacrylate, 2-hydroxyethyl acrylate, methyl acrylate, acrylamide, vinyl ether, vinyl acetate, divinylbenzene, ethylene glycol dimethacrylate, ethylene glycol diacrylate, pentaerythritol dimethacrylate, pentaerythritol diacrylate, N,N′

-methylenebisacrylamide, N,N′

-ethylenebisacrylamide, N,N′

-(1,2-dihydroxyethylene)bis-acrylamide, trimethylolpropane trimethacrylate and vinyl cyclodextrin.

9. The method of claim 1 wherein the composition further comprises a cross-linking agent.

10. The method of claim 1 in which the template moiety and the tail moiety are attached to one another via a linker.

11. The method of claim 1 wherein the tail moiety comprises a lipid or fatty acid.

12. The method of claim 1, wherein the template moiety corresponds to a portion of a macromolecule of interest.

13. The method of claim 12, wherein the template moiety corresponds to a terminal portion of the macromolecule.

14. The method of claim 12, wherein the macromolecule is a polynucleotide and the template moiety is an oligonucleotide.

15. The method of claim 12, wherein the macromolecule is a glycoprotein or polysaccharide and the template moiety is an oligosaccharide.

16. The method of claim 12, wherein the macromolecule is a polypeptide and the template moiety is a peptide.

17. The method of claim 16, wherein the sequence of the peptide corresponds to a contiguous sequence of the polypeptide.

18. The method of claim 17, wherein the peptide is between 3 and 15 amino acids in length.

19. The method of claim 18, wherein the peptide is between 4 and 15 amino acids in length.

20. The method of claim 19, wherein the peptide is between 4 and 7 amino acids in length.

21. The method of claim 16, wherein the portion of the polypeptide comprises the C-terminus of the polypeptide.

7. The method of claim 7 wherein the dispersant is selected from the group consisting of KLE 373 and polyisobutylene.

22. An imprint bead comprising a plurality of imprint cavities that are oriented and localized at or near the surface of the imprint bead.

23. The imprint bead of claim 22, wherein the imprint bead is between 0.1 μ

m and 600 μ

m in diameter.

24. The imprint bead of claim 23, wherein the imprint bead is between 0.5 μ

m and 600 μ

m in diameter.

25. The imprint bead of claim 24, wherein the imprint bead is between 0.1 μ

m and 16 μ

m in diameter.

26. The imprint bead of claim 24, wherein the imprint bead is between 100 μ

m and 600 μ

m in diameter.

27. The imprint bead of claim 23, wherein the imprint bead is between 0.1 μ

m and 10 μ

m in diameter.

29. The collection of claim 27, wherein the average diameter is between 0.5 μ

m and 600 μ

m.

31. The collection of claim 27, wherein the average diameter is between 0.1 μ

m and 16 μ

m.

28. A collection of imprint beads having an average diameter of the bead between 0.1 μ

m and 600 μ

m, wherein each imprint bead has a plurality of imprint cavities that are oriented and localized at or near the surface of the imprint bead.

30. The collection of claim 28, wherein the average diameter is between 100 μ

m and 600 μ

m.

32. The collection of claim 30, wherein the average diameter is between 0.1 μ

m and 10 μ

m.

33. A collection of imprint beads made by the process comprising the steps of:

(a) emulsifying a composition comprising a polar solvent, a non-polar solvent, an organic monomer which is polymerizable in aqueous solution and a conjugate molecule, said conjugate molecule comprising a template moiety and a tail moiety, to form an inverse emulsion, wherein the template moiety is capable of partitioning into the noncontinuous phase of the inverse emulsion and the tail moiety is capable of partitioning into the continuous phase of the inverse emulsion;

(b) polymerizing the organic monomer; and

(c) removing the conjugate molecule.