Analyte detection utilizing polynucleotide sequences, composition, process and kit

• US 9,134,302 B2
• Filed: 06/07/1995
• Issued: 09/15/2015
• Est. Priority Date: 05/05/1983
• Status: Active Grant

• Current Assignee: Enzo Diagnostics, Inc.
• Original Assignee: Enzo Diagnostics, Inc.
• Inventors: Pergolizzi, Robert G., Stavrianopoulos, Jannis G., Rabbani, Elazar, Engelhardt, Dean L., Kline, Stan, Olsiewski, Paula J.
• Primary Examiner(s): Brusca, John S
• Application Number:
  US08/479,995
• Publication Number:
  US 20100273145A1
• Time in Patent Office: 7,405 Days
• Field of Search: 435/5, 435/6, 435/810, 436/501, 536/23.1, 536/22.5, 536/24.5, 536 243- 2433, 935/77, 935/78
• US Class Current: 1/1
• CPC Class Codes: C12Q 1/6813 Hybridisation assays
• CPC Class Codes: C12Q 1/6876 Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
• CPC Class Codes: G01N 33/53 Immunoassay; Biospecific binding assay; Materials therefor
• CPC Class Codes: Y10T 436/143333 Saccharide [e.g., DNA, etc.]

Abstract

A method of detecting in a sample an analyte (A) having a molecularly recognizable portion thereon, which comprises: providing (B) a molecular bridging entity having thereon: (i) a portion capable of recognizing the molecularly recognizable portion on the analyte; and (ii) a portion comprising a polynucleotide sequence; and (C) a signalling entity having thereon: (i) a polynucleotide portion capable of annealing to the polynucleotide portion of the bridging entity, thereby to form a stable polynucleotide hybrid, and (ii) a signal generating portion; forming a complex comprising: (1) the analyte (A) complexed through its molecularly recognizable portion to (2) the recognizing portion of the entity (B); the entity (B) being complexed through the polynucleotide portion thereon to (3) the polynucleotide portion of the signalling entity; and detecting a signal by means of the signal generating portion present in the complex.

Claims

1. A polynucleotide sequence covalently attached to a monoclonal antibody.

2. A polynucleotide sequence covalently attached to a lectin.

3. A polynucleotide sequence covalently attached to a hormone receptor.

4. A polynucleotide sequence covalently attached to a hormone.

5. A circular DNA molecule covalently attached to a non-radiolabeled signal generating moiety that comprises an enzyme.

6. A circular DNA molecule comprising a polynucleotide that encodes part or all of a gene, wherein the DNA molecule is covalently attached to a non-radiolabeled signal generating moiety that comprises biotin.

7. A circular DNA molecule comprising a polynucleotide that encodes part or all of a gene, wherein the DNA molecule is covalently attached to a non-radiolabeled signal generating moiety that comprises an antibody.

8. A circular DNA molecule comprising a polynucleotide that encodes part or all of a gene, wherein the DNA molecule is covalently attached to a non-radiolabeled signal generating moiety that comprises a fluorogenic compound.