Fluorogenic semiconductor nanocrystals

Patent metadata

Field	Value
Publication number	US-10001474-B2
Application number	US-201214124354-A
Country	US
Kind code	B2
Filing date	Jun 7, 2012
Priority date	Jun 7, 2011
Publication date	Jun 19, 2018
Grant date	Jun 19, 2018

Abstract

Fluorogenic semiconductor nanocrystals and compositions thereof are provided herein, including kits, assay systems and methods for their preparation and use.

First claim

What is claimed is:

1. A method of activating a fluorogenic semiconductor nanocrystal, comprising:
a) providing a water-dispersible fluorogenic semiconductor nanocrystal in an aqueous medium, the aqueous medium comprising:
a fluorescent semiconductor nanocrystal, wherein the nanocrystal comprises a semiconductor core, and an external semiconductor shell layer disposed on the semiconductor core;
a hydrophilic layer on the surface of the external semiconductor shell layer that renders the nanocrystal water-dispersible; and
a quencher group associated with the external semiconductor shell layer, wherein the quencher group is a copper ion, wherein the fluorescent semiconductor nanocrystal in the absence of the quencher group has an initial fluorescence intensity (F 0 ), and the fluorogenic semiconductor nanocrystal has a quenched fluorescence intensity (F Q ), wherein F Q /F 0 is less than about 1.0; and
b) contacting the nanocrystal with an activator at a pH of greater than 7, wherein the activator comprises a cyanogenic enzyme substrate; and
c) contacting the cyanogenic enzyme substrate with a cyanogenic enzyme capable of cleaving the cyanogenic substrate to release a compound comprising a nitrile, an isonitrile or a cyanide group, wherein the fluorescence intensity of the fluorogenic semiconductor nanocrystal after contact with the activator increases, wherein the nanocrystal after contact with the activator has an activated fluorescence intensity (F A ), wherein F A /F Q is 10 or greater.

2. The method of claim 1 , wherein the cyanogenic enzyme substrate is selected from amygdalin, prunasin, dhurrin, vicianin, linamarin, lotaustralin, cyanohydrins, cyanohydrin esters, cyanohydrin amides, acetone cyanohydrins, mandelonitrile, and acyl cyanides.

3. The method of claim 1 , wherein the cyanogenic enzyme is a glucosidase.

4. The method of claim 1 , wherein the cyanogenic enzyme is selected from almond β-glucosidase, amygdalase, and linamarase.

5. The method of claim 1 , further comprising illuminating the fluorogenic semiconductor nanocrystal with light.

6. The method of claim 1 , wherein the semiconductor core comprises CdSe.

7. The method of claim 1 , wherein the external semiconductor shell layer comprises a semiconductor material selected from Group II-VI, Group II-II-VI, and Group II-VI-VI semiconductor materials.

8. The method of claim 1 , further comprising hydrolyzing the compound to release the isonitrile or cyanide group.

Assignees

• Welch Eric
• Bussian David
• Ignatius Michael
• Greenfield Lawrence
• Life Technologies Corp

Inventors

• Welch Eric
• Bussian David
• Ignatius Michael
• Greenfield Lawrence

Classifications

• G01N33/588 Primary

  with semiconductor nanocrystal label, e.g. quantum dots · CPC title

• C09K11/08

  containing inorganic luminescent materials · CPC title

• G01N33/581 Primary

  with enzyme label (including co-enzymes, co-factors, enzyme inhibitors or substrates) · CPC title

• B82Y15/00

  Nanotechnology for interacting, sensing or actuating, e.g. quantum dots as markers in protein assays or molecular motors · CPC title

• G01N33/525 Primary

  Multi-layer analytical elements · CPC title