Synthesis of fluorescent noble metal nanoparticles

• US 9,395,371 B2
• Filed: 09/23/2011
• Issued: 07/19/2016
• Est. Priority Date: 09/24/2010
• Status: Active Grant

• Current Assignee: The Governing Council of the University of Toronto (University of Toronto)
• Original Assignee: The Governing Council of the University of Toronto (University of Toronto)
• Inventors: Chan, Warren, Perrault, Steven, Chou, Leo
• Primary Examiner(s): CROW, ROBERT THOMAS
• Application Number:
  US13/825,771
• Publication Number:
  US 20130183665A1
• Time in Patent Office: 1,761 Days
• Field of Search: None
• US Class Current: 1/1
• CPC Class Codes: A61K 33/242 Gold; Compounds thereof
• CPC Class Codes: B22F 1/054 Nanosized particles
• CPC Class Codes: B82Y 30/00 Nanotechnology for materials or surface science, e.g. nanocomposites
• CPC Class Codes: B82Y 5/00 Nanobiotechnology or nanomedicine, e.g. protein engineering or drug delivery
• CPC Class Codes: C22C 5/02 Alloys based on gold
• CPC Class Codes: G01N 33/533 with fluorescent label
• CPC Class Codes: G01N 33/587 Nanoparticles

Abstract

A process for the production of fluorescent nanoparticles selected from noble metal or silica nanoparticles. Noble metals or silica nanoparticles provide a platform to which fluorophores and other detection molecules can be added. The fluorescent nanoparticles can be used as regents, diagnostic assays and in the tracking or targeting of tumors.

Claims

1. A process for the production of noble metal or silica nanoparticles as probes capable to fluoresce to label cells and tissues, the method comprising:

(1) providing a platform of the nanoparticles;

(2) covering the surfaces of the nanoparticles to saturation with thiol-terminated polymers by one of the following methods;

(a) mixing the nanoparticles with methoxy-(polyethylene glycol)-thiol, the methoxy-(polyethylene glycol)-thiol having a MW of about 10,000 atomic mass units (amu), and biotin-(polyethylene glycol)-thiol, the biotin-(polyethylene glycol)-thiol having a MW of about 10,000 amu;

(b) mixing the nanoparticles with fluorescently-labeled methoxy-(polyethylene glycol)-thiol, the methoxy-(polyethylene glycol)-thiol having a MW of about 10,000 amu and biotin-(polyethylene glycol)-thiol, the biotin-(polyethylene glycol)-thiol having a MW of about 10,000 amu;

(c) coordinating thiol and biotin thiol to the surfaces of the nanoparticles by a non-covalent bond; and

(d) directly conjugating methoxy-thiol and biotin-thiol to the surfaces of the nanoparticles,

whereby the polymers bind to the surfaces of the nanoparticles as a brush layer via thiol particle coordination of the thiol ends so that the biotin or methoxy ends are free, thereby obtaining polymer-covered nanoparticles;

(3) homogeneously mixing the resulting polymer-covered nanoparticles with fluorophore labeled avidin or an avidin derivative thereof in proportions of 1 biotin molecule for every 10 to 1000 avidin molecules thereby obtaining fluorescent multi-coloured nanoparticle-avidin complexes and the fluorophore labeled avidin or a derivative thereof being spaced away from the surface of the noble metal or silica nanoparticles, thus reducing or removing the potential quenching of the fluorophore.

2. The process as claimed in claim 1 wherein the fluorophore is an organic fluorophore selected from the group comprising 2-[5-(1-{6-[(2,5-dioxopyrrolidin-1-yl)oxy]-6-oxohexyl}-3,3-dimethyl-5-sulfonato-1,3-dihydro-2H-indol-2-ylidene)penta-1,3-dien-1-yl]-1-ethyl-3,3-dimethyl-3H-indolium-5-sulfonate and fluorescein isothiocyanate and wherein the organic fluorophore is coated onto the surfaces of the polymer coatings of the nanoparticles via a polyethylene-thiol spacer.

3. The process of claim 1 wherein the distance between the fluorophore to the surface of the nanoparticle is from about 4 to about 15 nm.

4. The process as claimed in claim 1 wherein the nanoparticles are noble metal nanoparticles.

5. The process as claimed in claim 1 wherein the process further comprises adding biotinylated targeting molecules to the nanoparticle-avidin complexes, and wherein the targeting molecules are selected from antibodies, peptides and aptamers.

6. A process for the production of gold nanoparticles as probes capable to fluoresce to label cells and tissues, the method comprising:

(1) providing a platform of gold nanoparticles;

(2) covering the surfaces of the gold nanoparticles to saturation with thiol-terminated polymers by one of the following methods;

(a) mixing the gold nanoparticles with methoxy-(polyethylene glycol)-thiol and biotin-(polyethylene glycol)-thiol;

(b) coordinating thiol and biotin thiol to the surfaces of the gold nanoparticles by a non-covalent bond; and

(c) directly conjugating methoxy-thiol and biotin-thiol to the surfaces of the gold nanoparticles,

whereby the polymers bind to the surfaces of the gold nanoparticles as a brush layer via thiol particle coordination of the thiol ends so that the biotin or methoxy ends are free, thereby obtaining polymer-covered gold nanoparticles; and

(3) homogeneously mixing the resulting polymer-covered gold nanoparticles with organic fluorophore labeled avidin or a derivative thereof in proportions of 1 biotin molecule for every 40 to 1000 avidin molecules thereby obtaining fluorescent multi-coloured gold nanoparticle-avidin complexes, fluorophore labeled avidin or a derivative thereof being spaced away from the gold nanoparticle surface, thus reducing or removing the potential quenching of the organic fluorophore.