SYSTEM AND METHOD FOR ITERATIVE DETECTION OF BIOLOGICAL MOLECULES

• US 20160054308A1
• Filed: 08/07/2015
• Published: 02/25/2016
• Est. Priority Date: 08/22/2014
• Status: Active Grant

• Current Assignee: Jia Guo
• Original Assignee: Jia Guo
• Inventors: Guo, Jia
• Granted Patent: US 9,933,431 B2
• Time in Patent Office: Days
• US Class Current: 1/1
• CPC Class Codes: C12Q 1/6823 Release of bound markers
• CPC Class Codes: C12Q 2527/125 Specific component of sample, medium or buffer
• CPC Class Codes: C12Q 2563/107 fluorescence
• CPC Class Codes: G01N 33/582 with fluorescent label

Abstract

A system for the iterative detection of biological molecules includes a probe, and a fluorophore tethered to the probe with an azide-based linker. The linker is configured to be cleaved in the presence of tris(2-carboxyethyl)phosphine (TCEP), and the on/off ratio between a signal measured before treatment with TCEP and a signal measured after treatment with TCEP is at least about 20:1.

Claims

1. A system for iterative detection of biological molecules, comprising:

a probe; and

a fluorophore tethered to the probe with an azide-based linker;

wherein the linker is configured to be cleaved in the presence of tris(2-carboxyethyl)phosphine (TCEP), and

wherein the on/off ratio between a signal measured before treatment with TCEP and a signal measured after treatment with TCEP is at least about 20;

1.

2. The system of claim 1 wherein the probe is selected from an oligonucleotide and an antibody.

3. The system of claim 1, wherein the linker is:

4. The system of claim 1, wherein the probe is selected for the detection of at least one of a DNA target, an RNA target, a protein target and a small molecule target.

5. The system of claim 1, wherein the linker includes a plurality of cleavage sites.

6. A method of detecting a biological molecule, comprising:

incubating a sample including a target molecule with a detection system, the detection system including a probe and a fluorophore tethered to the probe with an azide-based linker;

exciting the sample at a first wavelength;

detecting the emission of a second wavelength from the sample; and

treating the sample with tris(2-carboxyethyl)phosphine, thereby cleaving the fluorophore from the probe.

7. The method of claim 6, further including, repeating the steps of incubating, exciting and detecting.

8. The method of claim 6 wherein the probe is selected from an oligonucleotide and an antibody.

9. The method of claim 6, wherein the linker is:

10. The method of claim 6, wherein the probe is selected for the detection of at least one of a DNA target, an RNA target, a protein target and a small molecule target.

11. The method of claim 6, wherein the linker includes a plurality of cleavage sites.

12. The method of claim 6, further including binding the target molecule with the probe.

13. The method of claim 6, wherein the on/off ratio between a signal measured before treatment with TCEP and a signal measured after treatment with TCEP is at least about 20:

1.

14. The method of claim 6, wherein the on/off ratio between a signal measured before treatment with TCEP and a signal measured after treatment with TCEP is at least about 50:

1.