IP Library Granted Patent US 8,043,810
Granted Patent B2
US 8,043,810 · App. 11/775,740 · Granted Oct 25, 2011

Analyte detection using autocatalytic chain reactions

Assignee: Eagle Eye Research, Inc.
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Quick Facts
Patent No.
US 8,043,810
App. No.
11/775,740
Granted
Oct 25, 2011
Kind
B2
Abstract

Compositions and methods for detecting the presence of analytes employing autocatalytic chain reactions (ACR) having super linear kinetics for amplification of signal are disclosed.

Claims (25)

1. A method to determine the presence or absence of an analyte in a sample which method comprises

contacting said sample with

a nanometer scale switching device (NSD) comprised of a nucleic acid that is specific for, and activated by, the presence of the analyte such that the NSD will remain in its off state in the absence of the analyte and switch to its on state in the presence of the analyte,

components comprised of nucleic acids that are components of an autocatalytic chain reaction (ACR) process which process is initiated by activation of the NSD or directly by the analyte, and operates with superlinear kinetics to produce a product wherein each subsequent ACR reaction cycle produces more than one further product from each product previously formed with a rate of product growth increasing over time, and

an indicator (IDR) responsive to said product; and

detecting the presence or absence of any response of the IDR to said product as indicative of the presence or absence of the analyte,

wherein said ACR comprises an autocatalytic nucleic acid cleavage reaction,

wherein said autocatalytic nucleic acid reaction opens a covalently circularized ribozyme or deoxyribozyme achieving a linearized form.

2. The method of claim 1 , wherein the analyte effects an allosteric transformation in said NSD to release or expose a trigger that initiates the ACR.

3. The method of claim 2 , wherein the NSD is activated by a substance other than a nucleic acid or by a photon, an α particle or a β particle.

4. The method of claim 1 , wherein

said autocatalytic nucleic acid cleavage reaction opens a covalently circularized inactive form of a ribozyme or deoxyribozyme achieving a linearized active form of said ribozyme or deoxyribozyme, and

wherein the NSD comprises a second inactive form of said ribozyme or deoxyribozyme that is activated in the presence of analyte, and wherein the product is the linearized active ribozymes or deoxyribozymes, and

wherein all components are dissolved in solution phase and not attached to a solid support.

5. The method of claim 4 , wherein said analyte activates an intermediate that activates said second inactive form of said ribozyme or deoxyribozyme.

6. The method of claim 4 , wherein the ribozyme or deoxyribozyme is the 10-23 deoxyribozyme.

7. The method of claim 4 , wherein said IDR employs colorimetric detection wherein DNA labeled gold nanoparticles (DNA-AuNP) are added to the reaction after an appropriate time period such that the ACR proceeds sufficiently such that the DNA-AuNP do not halt the ACR prematurely.

8. A composition for performing the method of claim 1 , which composition comprises

covalently circularized inactive ribozymes or deoxyribozymes which are activated by linearization via backbone bond cleavage, wherein a linearized active form of said ribozymes or deoxyribozymes recognizes and opens said covalently circularized inactive ribozymes or deoxyribozymes in an ACR process that is characterized by superlinear kinetics, produces a product and is initiated by an activated NSD or by an analyte,

an IDR responsive to said product, and

optionally comprising an NSD specific for, and activated by, said analyte.

9. A kit for performance of the method of claim 1 containing, optionally in separate containers,

covalently circularized inactive ribozymes or deoxyribozymes which are activated by linearization via backbone bond cleavage, wherein a linearized active form of said ribozymes or deoxyribozymes recognizes and opens said covalently circularized inactive ribozymes or deoxyribozymes in an ACR process that is characterized by superlinear kinetics that produces a linearized product with a rate of product growth increasing over time and is initiated by an activated NSD or by an analyte,

an IDR responsive to said product, and

optionally an NSD specific for, and activated by, said analyte.

Assignments (7)
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Nov 12, 2008
From: LABEAN, THOMAS H.
To: EAGLE EYE RESEARCH, INC.
Reel/Frame 021822/0693 →
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Nov 12, 2008
From: YIN, PENG
To: EAGLE EYE RESEARCH, INC.
Reel/Frame 021822/0610 →
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Nov 12, 2008
From: SHETTY, GEETHA
To: EAGLE EYE RESEARCH, INC.
Reel/Frame 021822/0690 →
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Nov 12, 2008
From: REIF, JOHN H.
To: EAGLE EYE RESEARCH, INC.
Reel/Frame 021822/0713 →
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Nov 12, 2008
From: SCHULTES, ERIK A.
To: EAGLE EYE RESEARCH, INC.
Reel/Frame 021822/0724 →
CONFIRMATORY LICENSE Recorded Sep 4, 2008
From: EAGLE EYE, INC.
To: AFRL/RIJ
Reel/Frame 021477/0612 →
CONFIRMATORY LICENSE Recorded Aug 6, 2008
From: EAGLE EYE, INC.
To: AFRL/RIJ
Reel/Frame 021348/0216 →
Continuity (2)
Provisional Application 60915659 · May 2, 2007
Related Publication 20090087838A1 · Apr 2, 2009