US 6,750,016 B2 · Nanoparticles having oligonucleotides attached thereto and uses therefor

Abstract

The invention provides methods of detecting a nucleic acid. The methods comprise contacting the nucleic acid with one or more types of particles having oligonucleotides attached thereto. In one embodiment of the method, the oligonucleotides are attached to nanoparticles and have sequences complementary to portions of the sequence of the nucleic acid. A detectable change (preferably a color change) is brought about as a result of the hybridization of the oligonucleotides on the nanoparticles to the nucleic acid. The invention also provides compositions and kits comprising particles. The invention further provides methods of synthesizing unique nanoparticle-oligonucleotide conjugates, the conjugates produced by the methods, and methods of using the conjugates. In addition, the invention provides nanomaterials and nanostructures comprising nanoparticles and methods of nanofabrication utilizing nanoparticles. Finally, the invention provides a method of separating a selected nucleic acid from other nucleic acids.F

Claims41

Dependent claims

A method for detecting one or more target analytes in a sample comprising:

(a) providing at least one type of nanoparticle conjugates comprising
(i) nanoparticles having at least one type of first oligonucleotides bound thereto wherein the first oligonucleotides are attached to the nanoparticles in an aging process comprising contacting the oligonucleotides with the nanoparticles in aqueous solution for a period of time sufficient to allow some of the first oligonucleotides to bind to the nanoparticles;
and contacting the first oligonucleotides and nanoparticles in an aqueous salt solution for an additional period of time sufficient to enable additional first oligonucleotides to bind to the nanoparticles;
and
(ii) at least one type of second oligonucleotides, each type of second oligonucleotides having bound thereto specific binding pair complements of a specific target analyte, the second oligonucleotides having a sequence that is complementary to at least a portion of the first oligonucleotides bound to the nanoparticles and are bound to at least a portion of the first oligonucleotides bound to the nanoparticles by contacting the second oligonucleotides with the first oligonucleotides bound to the nanoparticles, said contacting occurring under conditions effective for the hybridization between the second oligonucleotides and the first oligonucleotides bound to the nanoparticles;
(b) contacting the sample with the nanoparticle conjugates under conditions effective to allow specific binding interactions between the target analyte and its specific binding complement bound to the nanoparticle conjugate;
and
(c) observing a detectable change brought about by the specific binding interaction of the target analyte and the specific binding complement of said analyte.

5 more elements

A method for detecting one or more target analytes in a sample comprising:

(a) providing one or more types of nanoparticle conjugates comprising:
(i) nanoparticles having at least one type of first oligonucleotides bound thereto, wherein the first oligonucleotides are attached to the nanoparticles in an aging process comprising contacting the first oligonucleotides with the nanoparticles in aqueous solution for a period of time sufficient to allow some of the first oligonucleotides to bind to the nanoparticles;
and contacting the first oligonucleotides and nanoparticles in an aqueous salt solution for an additional period of time sufficient to enable additional first oligonucleotides to bind to the nanoparticles;
(ii) at least one type of second oligonucleotides, each type of second oligonucleotides having bound thereto specific binding complements of a specific target analyte;
and
(iii) linker oligonucleotides having at least two portions, a first portion of the linker oligonucleonucleotide is bound to the first oligonucleotides bound to the nanoparticles and a second portion of the linker oligonucleotides is bound to the second oligonucleotides having bound thereto specific binding complements of a target analyte by contacting the linker oligonucleotides, the second oligonucleotides, and first oligonucleotides bound to the nanoparticle, the contacting occurring under conditions effective for hybridization of the linker oligonucleotides to the second oligonucleotides and hybridization of the linker olignonucleotides to the first oligonucleotides bound to the nanoparticles;
(b) contacting the sample with the nanoparticle conjugates under conditions effective to allow specific binding interaction between the target analytes and specific binding complements bound to the nanoparticle conjugates;
and
(c) observing a detectable change brought about by the specific binding interaction of the target analytes and the specific binding complements of said target analytes.

6 more elements

A method for detecting one or more target analytes comprising:

(a) providing (1) a type of nanoparticle conjugate comprising
(i) nanoparticles having at least one type of first oligonucleotides bound thereto wherein the first oligonucleotides are attached to the nanoparticles in an aging process comprising contacting the oligonucleotides with the nanoparticles in aqueous solution for a period of time sufficient to allow some of the first oligonucleotides to bind to the nanoparticles;
and contacting the first oligonucleotides and nanoparticles in an aqueous salt solution for an additional period of time sufficient to enable additional first oligonucleotides to bind to the nanoparticles;
and
(ii) at least one type of second oligonucleotides, each type of second oligonucleotides having bound thereto specific binding pair complements of a specific target analyte, the second oligonucleotides having a sequence that is complementary to at least a portion of the first oligonucleotides bound to the nanoparticles and are bound to at least a portion of the first oligonucleotides bound to the nanoparticles by contacting the second oligonucleotides with the first oligonucleotides bound to the nanoparticles, said contacting occurring under conditions effective for the hybridization between the second oligonucleotides and the first oligonucleotides bound to the nanoparticles;
(2) a support having at least one type of third oligonucleotides bound thereto, and
(b) modifying target analytes believed to be present in a sample by binding the target analytes to fourth oligonucleotides, the fourth oligonucleotide bound to the target analyte having a sequence that is complementary to at least a portion of the third oligonucleotides bound to the support;
(c) contacting the third oligonucleotides bound to the support with the fourth olignonucleotides bound to the target analytes under conditions effective to allow hybridization between the oligonucleotides bound to the support and the oligonucleotides bound to the target analytes;
(c) contacting the target analytes bound to the support with the nanoparticle conjugates under conditions effective to allow for specific binding interactions between the target analyte bound to the support and the specific binding complement bound to the nanoparticle conjugate;
and
(d) observing a detectable change dependent on the specific binding of the target analytes to the specific binding complement of the target analytes.

8 more elements

A method for detecting one or more target analytes in a sample comprising:

(a) providing (1) at least one type of nanoparticle conjugates comprising:
(i) nanoparticles having at least one type of first oligonucleotides bound thereto wherein the first oligonucleotides are attached to the nanoparticles in an aging process comprising contacting the oligonucleotides with the nanoparticles in aqueous solution for a period of time sufficient to allow some of the first oligonucleotides to bind to the nanoparticles;
and contacting the first oligonucleotides and nanoparticles in an aqueous salt solution for an additional period of time sufficient to enable additional first oligonucleotides to bind to the nanoparticles;
and
(ii) at least one type of second oligonucleotides, each type of second oligonucleotides having bound thereto specific binding pair complements of a specific target analyte, the second oligonucleotides having a sequence that is complementary to at least a portion of the first oligonucleotides bound to the nanoparticles and are bound to at least a portion of the first oligonucleotides bound to the nanoparticles by contacting the second oligonucleotides with the first oligonucleotides bound to the nanoparticles, said contacting occurring under conditions effective for the hybridization between the second oligonucleotides and the first oligonucleotides bound to the nanoparticles;
(2) a support having at least one type of third oligonucleotides bound thereto,
(3) linker oligonucleotides, wherein the linker oligonucleotides have a sequence with at least two portions, the third oligonucleotides bound to the support have a sequence that is complementary to the first portion of the linker oligonucleotide sequence, and
(b) modifying target analytes believed to be present in a sample by binding the target analytes to fourth oligonucleotides, the fourth oligonucleotide having a sequence that is complementary to the second portion of the linker oligonucleotides sequence;
(c) contacting the linker oligonucleotides with the third oligonucleotides bound to the support, said contacting occuring under conditions effective to allow hybridization between the oligonucleotides bound to the support with the first portion of the linker oligonucleotides;
(c) contacting the linker oligonucleotides with the fourth oligonucleotides bound to the target analytes under conditions effective to allow hybridization between the fourth oligonucleotides bound to the target analytes and the second portion of the linker oligonucleotides;
(d) contacting target analytes bound to the support with the nanoparticle conjugates under conditions effective to allow specific binding interaction between the target analytes bound to the support and the specific binding complements bound to the nanoparticle conjugates;
and
(e) observing a detectable change dependent on the specific binding of the target analytes to the specific binding complement of the target analytes.

10 more elements

A method for detecting one or more target analytes comprising:

(a) providing (1) a type of nanoparticle conjugates comprising
(i) nanoparticles having at least one type of first oligonucleotides bound thereto, wherein the first oligonucleotides are attached to the nanoparticles in an aging process comprising contacting the first oligonucleotides with the nanoparticles in aqueous solution for a period of time sufficient to allow some of the first oligonucleotides to bind to the nanoparticles;
and contacting the first oligonucleotides and nanoparticles in an aqueous salt solution for an additional period of time sufficient to enable additional first oligonucleotides to bind to the nanoparticles;
(ii) at least one type of second oligonucleotides, each type of second oligonucleotides having bound thereto a specific binding pair member complement of a specific target an analyte;
and
(iii) linker oligonucleotides having at least two portions, a first portion of the linker oligonucleonucleotides are bound to the first oligonucleotides bound to the nanoparticles and a second portion of the linker oligonucleotides are bound to the second oligonucleotides having bound thereto a specific binding complement of a target analyte by contacting the linker oligonucleotides, the second oligonucleotides, and first oligonucleotides bound to the nanoparticles, the contacting occurring under conditions effective for hybridization of the linker oligonucleotides to the second oligonucleotides and hybridization of the linker oligonucleotides to the first oligonucleotides bound to the nanoparticles;
(2) a support having third oligonucleotides bound thereto,
(b) modifying target analytes believed to be present in a sample by binding the target analytes to fourth oligonucleotides, the fourth oligonucleotide having a sequence that is complementary to at least a portion of the sequence of the third oligonucleotides bound the support;
(c) contacting the third oligonucleotides bound to the support with second oligonucleotides bound to a target analytes under conditions effective to allow hybridization between the third oligonucleotides bound to the support with the fourth oligonucleotides bound to the target analyte;
(d) contacting the target analytes bound to the support with the nanoparticle conjugates under conditions effective to allow specific binding interactions between the target analytes bound to the support and the specific binding complements bound to the nanoparticle conjugates;
and
(e) observing a detectable change dependent on the specific binding of the target analytes to the specific binding complements of the target analytes.

9 more elements

A method for detecting one or more target analytes comprising:

(a) providing (1) a support having one or more target analytes bound thereto, and (2) a nanoparticle conjugate comprising
(i) nanoparticles having at least one type of first oligonucleotides bound thereto wherein the first oligonucleotides are attached to the nanoparticles in an aging process comprising contacting the oligonucleotides with the nanoparticles in aqueous solution for a period of time sufficient to allow some of the first oligonucleotides to bind to the nanoparticles;
and contacting the first oligonucleotides and nanoparticles in an aqueous salt solution for an additional period of time sufficient to enable additional first oligonucleotides to bind to the nanoparticles;
and
(ii) at least one type of second oligonucleotides, each type of second oligonucleotides having bound thereto specific binding pair complements of a specific target analyte, the second oligonucleotides having a sequence that is complementary to at least a portion of the first oligonucleotides bound to the nanoparticles and are bound to at least a portion of the first oligonucleotides bound to the nanoparticles by contacting the second oligonucleotides with the first oligonucleotides bound to the nanoparticles, said contacting occurring under conditions effective for the hybridization between the second oligonucleotides and the first oligonucleotides bound to the nanoparticles;
(b) contacting a support having target analytes bound thereto with the nanoparticle conjugates under conditions effective to allow specific binding interactions between the target analytes bound to the support and specific binding complements bound to the nanoparticle conjugates;
(c) contacting the nanoparticle conjugates bound to the support with staining material to produce a detectable change;
and
(d) observing the detectable change.

6 more elements

Dependencies are inferred from the claim text.

Nanoparticles having oligonucleotides attached thereto and uses therefor

Life of this patent

Nanoparticles having oligonucleotides attached thereto and uses therefor

Life of this patent

Abstract

Claims41