Functionalized fluorescent nanocrystal compositions and methods for their preparation

• US 8,283,179 B2
• Filed: 03/13/2007
• Issued: 10/09/2012
• Est. Priority Date: 09/17/2001
• Status: Active Grant

• Current Assignee: Life Technologies Corporation (Thermo Fisher Scientific Incorporated)
• Original Assignee: Life Technologies Corporation (Thermo Fisher Scientific Incorporated)
• Inventors: Naasani, Imad
• Primary Examiner(s): Chevalier, Alicia
• Assistant Examiner(s): PATEL, RONAK C
• Application Number:
  US11/685,343
• Publication Number:
  US 20100291702A1
• Time in Patent Office: 2,037 Days
• Field of Search: 428402-407, 436/172
• US Class Current: 436/172
• 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: G01N 33/588 with semiconductor nanocrystal label, e.g. quantum dots
• CPC Class Codes: Y10T 428/265 1 mil or less
• CPC Class Codes: Y10T 428/2982 Particulate matter [e.g., sphere, flake, etc.]
• CPC Class Codes: Y10T 428/2989 Microcapsule with solid core [includes liposome]
• CPC Class Codes: Y10T 428/2991 Coated
• CPC Class Codes: Y10T 428/2998 including synthetic resin or polymer

Abstract

The present invention provides for functionalized fluorescent nanocrystal compositions and methods for making these compositions. The compositions are fluorescent nanocrystals coated with at least one material. The coating material has chemical compounds or ligands with functional groups or moieties with conjugated electrons and moieties for imparting solubility to coated fluorescent nanocrystals in aqueous solutions. The coating material provides for functionalized fluorescent nanocrystal compositions which are water soluble, chemically stable, and emit light with a high quantum yield and/or luminescence efficiency when excited with light. The coating material may also have chemical compounds or ligands with moieties for bonding to target molecules and cells as well as moieties for cross-linking the coating. In the presence of reagents suitable for reacting to form capping layers, the compounds in the coating may form a capping layer on the fluorescent nanocrystal with the coating compounds operably bonded to the capping layer.

Claims

1. A functionalized fluorescent nanocrystal comprising:

a fluorescent nanocrystal; and

a coating material, wherein the coating material comprises an imidazole-containing compound comprising two or more amino acids joined together.

2. The functionalized fluorescent nanocrystal of claim 1, wherein the fluorescent nanocrystal emits light with a quantum yield of greater than about 10%.

3. The functionalized fluorescent nanocrystal of claim 1, wherein the fluorescent nanocrystal emits light with a quantum yield of greater than about 50%.

4. The functionalized fluorescent nanocrystal of claim 1, wherein the fluorescent nanocrystal is soluble in a fluid with a surface energy of greater than 35 dynes/cm.

5. The functionalized fluorescent nanocrystal of claim 1, wherein the fluorescent nanocrystal is soluble in an aqueous solution.

6. The functionalized fluorescent nanocrystal of claim 1, wherein the coating material is bound to a molecular probe.

7. The functionalized fluorescent nanocrystal of claim 6, wherein the molecular probe is a polynucleotide of known sequence.

8. The functionalized fluorescent nanocrystal of claim 6, wherein the molecular probe is selected from the group consisting of avidin, ConA, lectin, monoclonal antibodies, peptides, aptamers, and nucleic acid molecules, and nucleobases, and IgG.

9. The functionalized fluorescent nanocrystal of claim 1, wherein the nanocrystal is a doped metal oxide nanocrystal.

10. The functionalized fluorescent nanocrystal of claim 9, wherein the doped metal oxide nanocrystal is magnetized when irradiated with light.

11. The functionalized fluorescent nanocrystal of claim 1, wherein the fluorescent nanocrystal has a characteristic spectral emission intensity which is enhanced by the coating material.

12. The functionalized fluorescent nanocrystal of claim 1, wherein the fluorescent nanocrystal comprises a core material and a capping layer.

13. The functionalized fluorescent nanocrystal of claim 12, wherein the coating material is operably bound to the capping layer.

14. The functionalized fluorescent nanocrystal of claim 1, wherein the coating material further comprises a phosphine cross-linking compound.

15. The functionalized fluorescent nanocrystal of claim 14, wherein the phosphine cross-linking compound is an alkyl phosphine-containing compound.

16. The functionalized fluorescent nanocrystal of claim 14, wherein the phosphine cross-linking compound is tris (hydroxy methyl) phosphine or beta -[tris(hydroxymethyl)phosphino] propionic acid.

17. The functionalized fluorescent nanocrystal of claim 1, wherein at least one of the two or more amino acids in the imidazole-containing compound are selected from histidine, anserine, baleine, 5-amino-4-imidazolecarboxamide, 2-mercaptoimidazole, boc-histidine hydrazide, histidinol, 1-methylhistidine, 3-methylhistidine, imidazolysine, imidazole-containing ornithine, imidazole-containing alanine, carcinine, and histamine.

18. The functionalized fluorescent nanocrystal of claim 1, wherein the imidazole-containing compound is selected from carnosine, homocarnosine, histidylphenylalanine, cyclo-histidylphenylalanine, and histidylleucine.

19. A method for detecting a target species, the method comprising:

exciting a complex formed by operably bonding a fluorescent nanocrystal to a target molecule to form a complex wherein the fluorescent nanocrystal includes a coating material comprising an imidazole-containing compound, wherein the compound comprises two or more amino acids joined together; and

detecting the light emitted from the complex.

20. The method of claim 19, wherein the detecting further comprises detecting the intensity of light emitted from the complex in the perpendicular and parallel directions.

21. A process for the preparation of a functionalized nanocrystal comprising contacting a fluorescent nanocrystal comprising a core material and a capping layer with an imidazole-containing compound, wherein the compound comprises two or more amino acids joined together, to form a coating material on the fluorescent nanocrystal, wherein the compound is operably bound to the capping layer.

22. The process of claim 21, further comprising contacting the fluorescent nanocrystal with a phosphine cross-linking compound.