Design of labels for detection with a surface-seletive nonlinear optical technique

• US 20040146460A1
• Filed: 03/17/2004
• Published: 07/29/2004
• Est. Priority Date: 07/17/2001
• Status: Abandoned Application

• Current Assignee: Biodesy, LLC
• Original Assignee: Biodesy, LLC
• Inventors: Salafsky, Joshua S
• Application Number:
  US10/467,098
• Publication Number:
  US 20040146460A1
• Time in Patent Office: Days
• US Class Current: 424/9.6
• CPC Class Codes: B82Y 15/00 Nanotechnology for interacting, sensing or actuating, e.g. quantum dots as markers in protein assays or molecular motors
• CPC Class Codes: B82Y 30/00 Nanotechnology for materials or surface science, e.g. nanocomposites
• CPC Class Codes: G01N 27/44726 using specific dyes, markers or binding molecules
• CPC Class Codes: G01N 33/58 involving labelled substances G01N33/53 takes precedence

Abstract

Second harmonic, sum and difference frequency generation can be used to detect a variety of processes, which are otherwise undetectable, using nonlinear-active labels. The labels must have as high a hyperpolarizability as possible. Several designs for large hyperpolarizability second-harmonic active labels are described herein. These labels can be attached to any target molecule or particle, resulting in more highly nonlinear-optically active targets.

Claims

1. Nonlinear-active labels, said labels capable of being attached to a target for the purpose of studying said target using a surface-selective nonlinear optical technique.

2. The label according to claim 1, wherein the label includes a moiety selected from the group consisting of:

Oxazole or oxadizole molecules

5-aryl-2-(4-pyridyl)oxazole

2-aryl-5-(4-pyridyl)oxazole

2-(4-pyridyl)cycloalkano[d]oxazoles

Merocyanines

Stilbenes

Indodicarbocyanines

Hemicyanines

Stilbazims

Azo dyes

Cyanines

Stryryl-based dyes

Methylene blue

Diaminobenzene compounds

Polyenes

Diazostilbenes

Tricyanovinyl aniline

Tricyanovinyl azo

Melamines

Phenothiazine-stilbazole

Polyimides

Sulphonyl-substituted azobenzenes

Indandione-1,3-pyidinium betaine

Fluoresceins

Benzooxazoles

Perylenes

Polymethacrylates

Oxonols

Thiophenes

Bithiophenes

3. The label of claim 1, which includes an oxazole moiety based on a 1,3-oxazole.

4. The label of claim 2, wherein the oxazole moiety is a quaternary salt.

5. The label of claim 2, wherein the oxazole moiety is 2-(4-N-methylpyridinium)-4,5-dihydronaphtho [2,1-d]-1,3-oxazole p-toluenesulfonate.

6. The label of claim 2, wherein the oxazole is 2-(4-N-methylpyridinium)-4,5-dihydro6-methoxynaphtho[2,1-d]-1,3-oxazole p-toluenesulfonate.

7. The label of claim 2 wherein the oxazole moiety is a 5(2)-Aryl-2(5)-(4-pyridyl)oxazole

8. The label of claim 2 wherein the oxazole moiety is a 2,5-Diaryl-1,3-oxazole

9. The label of claim 1 wherein the label is:

1-(3-(succinimidyloxycarbonyl) benzyl)-4-(5-(4-methoxyphenyl) oxazol-2-yl)pyridinium bromide

10. The label of claim of 1 wherein the label is:

1-(2,3-epoxypropyl)-4-(5-(4-methoxyphenyl)oxazol-2-yl) pyridinium trifluoromethanesulfonate (PyMPO epoxide).

11. The label of claim 1 wherein the nonlinear-active label includes a non-centrosymmetric metallic or semiconductor particle.

12. The label of claim 1, wherein the nonlinear-active label includes a centrosymmetric metallic or semiconductor particle with a size of greater than or equal to 10% of the wavelength of the fundamental light.

13. The label of claim 1 which comprises bacteriorhodoposin or the green fluorescent protein.

14. The label of claim 1 including a genetically engineered mutant of bacteriorhodopsin or the green fluorescent protein.

15. The label of claim 1 wherein a target object is first derivatized with a linker and then coupled to the label.

16. The label according to claims 2-14 wherein the nonlinear active moieties, particles or proteins are coupled to a linker molecule.

18. The label of claim 17 wherein the nonlinear-active components include one or more of the following:

a moiety from claims 2-10, a particle from claims 11 and 12 or a protein from claims 13 and 14.

19. The label of claim 18 wherein the components include a linker molecule for attachment to the solid surface.

22. The label of claim 18 wherein the components include a linker molecule for attachment to a target.

29. The label of claims 19-21 wherein the linker is longer than 8 carbon-atom lengths.

30. The claims of 11-12 wherein the particle comprises Au, Ag, Pt, CdS, CdSe, TiO₂, GaAs, InP, GaP.

31. The claims of 11-12 wherein the particles are also complexed to or attached to any of the moieties, particles or proteins in claims 2-14.

32. The label of claim 19 wherein the nonlinear-active particle is attached to the solid surface using functionalized alkylthiols.

40. The lable of claim 32 and 39, wherein the alkylthiols are functionalized with groups that are reactive toward amine, sulfhydryl, carboxylic, aldehyde, ketone, vicinal diol, glutamine, oligosaccharide, guadinium, NHS ester, methyl ester, hydroxyl, azido-methylcoumarin, Sulfo-NHS ester, maleimide, iodoacetyl, vinyl sulfone, —

CH bonds, carbodiimide-activated carboxyl, biotin, streptavidin, and phosphatidylcholine moieties.

44. The label of claim 42 wherein the chains are composed of any of the moieties, particles or proteins in claims 2-14.

50. A method for detecting a target at an interface of interest, said method comprising attaching a nonlinear active label according to any of claims 1-49 to said target and measuring said target at said interface using a surface-selective nonlinear optical technique.

51. The method of claim 50 wherein the interface is an air-water interface, a glass water interface, a solid-water interface or a solid-air interface, a vapor-liquid interface, a solid-solid-interface, or a liquid-liquid interface, cellular interface, membrane interface.

52. The method of claim 50 or 51 wherein the target is a protein, oligosaccharide, peptide, nucleic acid, liposome, small molecule, oligonucleotide, liposome, or biological cell, oligosaccharide, antibody, antigen, peptide, virus, receptor, drug, enzyme, ligand, carbohydrate.

53. The method of any of claims 50-52 wherein the nonlinear active technique is second harmonic generation, sum frequency generation or difference frequency generation.

54. The method of any of claims 50-53 wherein the targets are derivatized with biotin molecules and the nonlinear active labels are derivatized with streptavidin.

55. The labels according to any of claims 1-49 which are derivatized with biotin.

56. The method of any of claims 50-55 wherein said label is according to any of claims 19-22, 33 and 36 wherein the linkers include one or both of the following:

streptavidin, biotin.

57. The labels according to any of claims 5,16,19-22,29,36 and 54 wherein the linkers are functionalized with groups that are reactive toward amine, sulfhydryl, carboxylic, aldehyde, ketone, vicinal diol, glutamine, oligosaccharide, guanidium NHS ester, methyl ester, hydroxl, azido-methylcoumarin, Sulfo-NHS ester, maleimide, iodoacetyl, vinyl sulfone, —

CH bonds, carbodiimide-activated carboxyl, biotin, streptavidin, and phosphatidylcholine moieties.

58. The label according to any of claims of 15,16,19-22,29,36 and 54 wherein the linkers contain a functional group which is reactive towards a target, a second linker, a surface layer or a solid object.

60. The label according to any of claims 2-14 or 33 wherein the nonlinear active moiety, molecule or particle contains a functional group which is reactive towards a target, a linker, a surface layer or a solid object

62. The label according to claim 1, comprised of two or more distinct kinds of species comprising a single label, wherein the effect of the first kind of said species is to resonantly enhance the nonlinear activity of the second kind of said species.

63. The label according to claim 62, wherein the first kind of species are metallic or semiconductor particles and are used to create the resonance enhancement effect.

64. The label according to claim 62, wherein the metallic or semiconductor particles are centrosymmetric or non-centrosymmetric.

65. The label according to claim 62, wherein the second kind of species is any optically nonlinear active moiety, molecule or particle.

66. The label according to claim 62, wherein the two kinds of species are chemically bonded, attached, or linked to each other.

67. The label according to claim 66, wherein the average distance between the two kinds of species is of order angstroms or nanometers.

17. A non-linear active label capable of attachment to a target for the purpose of studying the target using a surface-selective nonlinear optical technique, wherein said label comprises a solid object to be used as a scaffold which provides a surface area onto which is attached at least one nonlinear-active component.

20. The label of claim 17 wherein a linker molecule is attached directly to the surface of the solid object, said linker allowing the coupling of the label to the target.

21. The label of claim 17 wherein a linker molecule is attached to the solid surface via a derivatized surface layer, said layer being attached to the surface, said linker allowing coupling of the layer to the target.

23. The label of claim 17 wherein the solid object is derivatized with a surface layer.

24. The claims of 17 and 18 wherein the nonlinear active component is attached to the surface layer.

25. The claims of 21 and 23 wherein the surface layer is a self-assembled monolayer

26. The label of claim 25 wherein the surface layer is in the chemical family of silane compounds.

27. The label of claim 17, wherein the solid object is a particle, cluster, colloidal particle, nanocrystal or nanoparticle of size scale ranging from nanometers to microns.

28. The label of claim 17, wherein the solid object is composed of a polymer, latex, polystyrene, silica, glass or silicon.

59. The label according to claim 17 wherein the solid object contains a surface functional group which is reactive towards a target, a linker, a surface layer or other solid objects.

61. The label according to claims 17, 27 or 28 wherein the solid object is spherical in shape, bead-like or quasi-spherical.

33. A non-linear label for use in detection of a target at an interface using a nonlinear optical technique, said label comprising a metallic or semiconductor particle.

34. The label of claim 33, wherein the particle is non-centrosymmetric.

35. The label of claim 33, wherein the particle is centrosymmetric and has a size of greater than or equal to 10% of the wavelength of the fundamental light used in said nonlinear optical technique.

36. The label of claim 33, wherein the particle is chemically derivatized with a linker molecule, said linker being reactive with a target object.

37. The label of claim 33, wherein the particle is chemisorbed or physisorbed to the target.

38. The label of claim 33, wherein the particle is composed of Au, Ag, Pt, CdS, CdSe, TiO₂, GaAs, InP, GaP.

39. The label of claim 33, wherein the metallic particle is derivatized with functionalized alkylthiols.

41. The label of claim 33, wherein the label consists of two or more particles in a composite, said particles being aggregated, adsorbed to each other, or attached to each other via covalent, noncovalent or electrostatic means, said composite being overall non-centrosymmetric.

42. A nonlinear active label for use in labeling a target and detecting said target using a surface-selective nonlinear optical technique, wherein said label comprises a polymeric molecule composed of two or more nonlinear-active moieties in which the moieties are coupled to each other in a linear head-to-tail fashion, and in which the orientation of each moiety in the chain is in the same direction.

43. The label of claim 42 wherein the chains are composed of nonlinear-active molecules.

45. A non-linear active label comprising a nonlinear-active protein.

46. The label of claim 45 wherein the target is a protein and the nonlinear-active label is a fusion protein.

47. The label of claim 45 wherein said target is a protein and the nonlinear-active label is fused to said target via co-expression of the genes encoding both proteins.

49. The label of claim 46 wherein the nonlinear-active protein is a genetically engineered mutant of either bacteriorhodopsin or the green fluorescent protein.

48. The label of 46 wherein the nonlinear-active protein is either bacteriorhodoposin or the green fluorescent protein.