LABELLED SILICA-BASED NANOMATERIAL WITH ENHANCED PROPERTIES AND USES THEREOF

• US 20130064776A1
• Filed: 10/11/2010
• Published: 03/14/2013
• Est. Priority Date: 10/09/2009
• Status: Active Grant

• Current Assignee: Bio-Rad Laboratories Incorporated
• Original Assignee: Raindance Technologies Incorporated (Bio-Rad Laboratories Incorporated), Centre National De La Recherche Scientifique, Universite De Strasbourg (Financière Meeschaert SA)
• Inventors: Griffiths, Andrew David, El Harrak, Abdeslam, Goust, Victoire, Mangeat, Thomas
• Granted Patent: US 10,520,500 B2
• Time in Patent Office: Days
• US Class Current: 424/9.6
• CPC Class Codes: G01N 33/54346 Nanoparticles
• CPC Class Codes: G01N 33/552 Glass or silica
• CPC Class Codes: G01N 33/582 with fluorescent label
• CPC Class Codes: Y10T 428/2993 Silicic or refractory material containing [e.g., tungsten oxide, glass, cement, etc.]

Abstract

The present invention relates to labelled silica-based nanoparticles with enhanced properties, to process for preparing them and to uses thereof.

Claims

1-44. -44. (canceled)

45. A nanoparticle comprising a silica core, having a diameter of about 200 nanometers or less, and having a label bound to the silica.

46. The nanoparticle of claim 45, further comprising a shell surrounding the core.

47. The nanoparticle of claim 46, wherein the shell comprises silica.

48. The nanoparticle of claim 45, further comprising a second label bound to the core.

49. The nanoparticle of claim 46, wherein the shell comprises a second label.

50. The nanoparticle of claim 45, wherein the first label is a fluorescent label having a tunable fluorescence polarization.

51. The nanoparticle of claim 49, wherein the nanoparticle has a tunable fluorescence polarization.

52. The nanoparticle of claim 45, wherein the first label comprises an organic molecule, an organic dye, an inorganic dye, an inorganic molecule, a magnetic particle or a radioactive compound.

53. The nanoparticle of claim 45, wherein the first label is a fluorescent dye or phosphorescent dye.

54. The nanoparticle of claim 45, wherein the nanoparticle has a diameter of about 25 nanometers or less.

55. The nanoparticle of claim 54, wherein the nanoparticle has a diameter of between about 2 nanometers and about 15 nanometers.

56. The nanoparticle of claim 46, wherein the nanoparticle comprises an additional layer of functionalizing molecules or is grafted with functionalizing molecules.

57. The nanoparticle of claim 56, wherein the functionalizing molecules are selected from the group consisting of silanes, maleimides, thiols, amines functions, polymers, proteins, antibodies, antigens, sugars, PEGs, organic molecules, and enzymes.

58. A method for making a nanoparticle comprising a core and a first label, comprising the steps of:

(a) providing a first label bound to a first molecule comprising silane thereby forming a silane functionalized label;

(b) providing a first solution comprising free silicon-containing molecules;

(c) mixing the silane functionalized label and the first solution to form a first mixed solution;

(d) reducing the pH of the first mixed solution thereby allowing conditions for the formation of covalent bonds among the silicon-containing molecules to form silica within which the first label is covalently bound, thereby nucleating the core; and

(e) allowing sufficient time for the core to grow until stopped.

59. The method of claim 58, wherein step (d) includes adding an ion-exchange resin to the first mixed solution, thereby to reduce the pH.

60. The method of claim 58, wherein the silane comprises APTES, APTMS, MPTMS, or MPTES.

61. The method of claim 58, wherein the method further comprises:

(f) mixing the grown core with a second solution comprising free silicon-containing molecules to form a second mixed solution;

(g) reducing the pH of the second mixed solution thereby growing a shell; and

(h) allowing sufficient time for the shell to grow until stopped.

62. The method of claim 61, wherein the second solution further comprises a second label.

63. The method of claim 61, wherein steps (f) through (h) are repeated one or more times, and wherein the second solution comprises a label different than the first or preceding labels during each repeat.

64. The method of claim 61, further comprising the steps of:

(i) optionally adding a functionalizing agent surrounding the shell to form a functionalized shell; and

(j) tuning one or more properties of the nanoparticle by grafting one or more surface molecules to the shell or, if present to the functionalized shell.

65. The method of claim 64, wherein the functionalizing agent comprises silanes, maleimides, thiols, or amines.

66. The method of claim 64, wherein the surface molecules comprise a polymer, protein, antibody, antigen, sugar, PEG, organic molecule, or enzyme.

67. The method of claim 58, wherein growth of the core or the shell is stopped by depletion of a constituent, addition of a quenching reagent, changing temperature, or changing pH.

68. A method of detecting a biological entity, comprising the steps of:

(a) attaching a nanoparticle to the biological entity via surface functionalizing molecules, wherein the nanoparticle comprises a silica core, has a diameter of about 200 nanometers or less, and has a label bound to the silica; and

(b) detecting the label.