Functionalization of and use of functionalized second harmonic generating nanoprobes

• US 9,221,919 B2
• Filed: 11/21/2012
• Issued: 12/29/2015
• Est. Priority Date: 11/21/2011
• Status: Active Grant

• Current Assignee: California Institute of Technology
• Original Assignee: California Institute of Technology
• Inventors: Pantazis, Periklis, Culic-Viskota, Jelena, Dempsey, William P., Fraser, Scott E.
• Primary Examiner(s): Dickinson, Paul
• Application Number:
  US13/683,975
• Publication Number:
  US 20130129628A1
• Time in Patent Office: 1,133 Days
• US Class Current: 1/1
• CPC Class Codes: A61K 49/0063 characterised by a special physical or galenical form, e.g. emulsions, microspheres
• CPC Class Codes: A61K 49/0093 Nanoparticle, nanocapsule, nanobubble, nanosphere, nanobead, i.e. having a size or diameter smaller than 1 micrometer, e.g. polymeric nanoparticle
• CPC Class Codes: C07F 7/10 containing nitrogen having a Si-N linkage
• CPC Class Codes: C07F 7/1804 Compounds having Si-O-C linkages Si-O-acyl linkages C07F7/1896
• CPC Class Codes: C07F 7/28 Titanium compounds
• CPC Class Codes: C07K 17/14 Peptides being immobilised on, or in, an inorganic carrier
• CPC Class Codes: C08G 65/33303 containing amino group
• CPC Class Codes: C08G 65/48 Polymers modified by chemical after-treatment
• CPC Class Codes: G01N 33/588 with semiconductor nanocrystal label, e.g. quantum dots
• CPC Class Codes: Y10S 977/773 Nanoparticle, i.e. structure having three dimensions of 100 nm or less
• CPC Class Codes: Y10S 977/902 Specified use of nanostructure

Abstract

Functionalized second harmonic nanoprobes for imaging samples and a method of using such probes to monitor the dynamics different processeses using a variety of imaging techniques are provided. The functionalized second harmonic generating (SHG) nanoprobes are comprised of various kinds of nanocrystalline materials that do not possess an inversion symmetry and therefore are capable of generating second harmonic signals that can then be detected by conventional two-photon microscopy, and are provided with functional surface modifications that allow for targeted imaging of a variety of biological and non-biological processes and structures such as cell signaling, neuroimaging, protein conformation probing, DNA conformation probing, gene transcription, virus infection and replication in cells, protein dynamics, tumor imaging and cancer therapy evaluation and diagnosis as well as quantification in optical imaging.

Claims

1. A functionalized probe nanostructure comprising:

a nanocrystalline body capable of exposing free surface hydroxyls defining an outer surface and being formed of a crystal material, the crystal material having an internal crystal structure having a unit cell having no inversion symmetry such that the crystal material generates a second harmonic signal independent of the overall shape of the nanocrystalline body when illuminated by an external excitation source,

a functionalization platform comprising a plurality of modifying molecules having amine terminal groups, the modifying molecules being linked to the outer surface of the nanocrystalline body through a plurality of hydroxyl molecules; and

a plurality of functionalization molecules bound to the amine terminal groups of the functionalization platform.

2. The probe nanostructure of claim 1, wherein the crystal material of the nanocrystalline body is selected from the group consisting of BaTiO₃, SiC, ZnO, LiNbO₃, KNbO₃, KTiOPO₄, Fe(IO₃)₃, N-(4-nitrophenyI)-(L)-prolinol, urea, 4-Nitroaniline, 2-Methyl-4-nitroaniline, 3-Methyl-4-methoxy-4′

-nitrostilbene), β

-BaB₂O₄, LiB₃O₅, KH₂PO₄, NH₄H₂PO₄, KD₂PO₄, CsLiB₆O₁₀, KTiOAsO₄, LiTaO₃, RbTiOAsO₄, BiB₃O₆, K₂Al₂B₂O₇, KBe₂BO₃F₂, BaAlBO₃F₂, La₂CaB₁₀O₁₉, GdCa₄O(BO₃)₃, YCa₄O(BO₃)₃, Li₂B₄O₇, LiRbB₄O₇, RbTiOPO₄, KB₅O₈.4H₂O, CsB₃O₅, C₄H₇D₁₂N₄PO₇, a-HIO₃, LiCOOH.H₂O, CsH₂AsO₄, CsD₂AsO₄, RbH₂PO₄, CsTiOAsO₄, Ba₂NaNb₅O₁₅, K₃Li₂Nb₅O₁₅, CO(NH₂)₂, and LiIO₃.

3. The probe nanostructure of claim 1, wherein the functionalization platform is formed from an amine functionalized reagent selected from the group consisting of N-aminoethyl-2,2,4-trimethyl-1-aza-2-silacyclopentane, 3-aminopropyliriethoxysilane, and 3-aminopropylirimethoxysilane, N-succinimidyl S-acetylthioacetate, isothiocyanates, acyl azides, N-hydroxysuccinimide esters, sulfonyl chloride derivatives, epoxides, fluorobenzene derivatives, and carbonate compounds.

5. The probe nanostructure of claim 1, wherein the functionalization molecule is selected from the group of a non-bio-reactive coating and a bio-reactivity enhancer.

6. The probe nanostructure of claim 5, wherein the bio-reactivity enhancer is a PEG analogue selected from the group consisting of biotin, bromo, amino, aminoethyl, carboxymethyl, succinimidosuccinate, tosylate, mesylate, aldehyde, actadecylamine, monopalmitate, and stearoyloxy derivatives of PEG.

7. The probe nanostructure of claim 1, wherein the functionalization molecule is a glycosylated antibody originating from a host.

8. The probe nanostructure of claim 1, wherein the functionalization molecule is configured to target or label a specific cell, biological molecule or molecule.

9. The probe nanostructure of claim 1, wherein the functionalized probe nanostructure is configured to be acceptable for use in conjunction with a technique selected from the group consisting of SHG imaging, direct nucleic acid sequencing in a Multi-SHG Detection Imaging modality, FRESH, optical monitoring of electric fields, imaging/detecting medical conditions or neoplasm, detection or tracking of therapeutic agents, and rapid detection systems.

10. The probe nanostructure of claim 1, wherein the functionalized probe nanostructure includes a plurality of different biologically relevant functionalization molecules.

11. The probe nanostructure of claim 1, wherein the functionalized probe nanostructure further includes at least one additional linkage between the functionalization platform and a biologically relevant functionalization molecule.

4. The probe nanostructure of dam 1, wherein the functionalization molecules are bound to the functionalization platform through a linkage selected from the group consisting of mono functional or multifunctional PEGylation, biocompatible polymers, click chemistry, and antibody targeting.