Compositions for detection and analysis of polynucleotides using light harvesting multichromophores

• US 9,371,559 B2
• Filed: 01/23/2012
• Issued: 06/21/2016
• Est. Priority Date: 06/20/2002
• Status: Active Grant

• Current Assignee: Regents of the University of California (University of California)
• Original Assignee: Regents of the University of California (University of California)
• Inventors: Bazan, Guillermo C., Gaylord, Brent S.
• Primary Examiner(s): Gross, Christopher M
• Application Number:
  US13/356,500
• Publication Number:
  US 20130190193A1
• Time in Patent Office: 1,611 Days
• Field of Search: None
• US Class Current: 1/1
• CPC Class Codes: B82Y 15/00 Nanotechnology for interacting, sensing or actuating, e.g. quantum dots as markers in protein assays or molecular motors
• CPC Class Codes: C12Q 1/6818 involving interaction of two or more labels, e.g. resonant energy transfer
• CPC Class Codes: C12Q 2525/107 incorporating a peptide nucleic acid
• CPC Class Codes: C12Q 2527/125 Specific component of sample, medium or buffer
• CPC Class Codes: C12Q 2565/107 Alteration in the property of hybridised versus free label oligonucleotides

Abstract

Methods, compositions and articles of manufacture for assaying a sample for a target polynucleotide are provided. A sample suspected of containing the target polynucleotide is contacted with a polycationic multichromophore and a sensor PNA complementary to the target polynucleotide. The sensor PNA comprises a signaling chromophore to absorb energy from the excited multichromophore and emit light in the presence of the target polynucleotide. The methods can be used in multiplex form. Kits comprising reagents for performing such methods are also provided.

Claims

1. A composition comprising:

a water soluble polycationic multichromophore conjugated polymer, comprising a backbone of conjugated segments, and

a signaling chromophore,

wherein the composition is in an aqueous solution, and

wherein a greater amount of energy is emitted from the signaling chromophore upon excitation of the conjugated polymer with incident light than from direct excitation of the signaling chromophore with incident light.

2. The composition of claim 1, wherein the aqueous solution comprises a biological sample.

3. The composition of claim 1, wherein the conjugated polymer transfers energy to the signaling chromophore.

4. The composition of claim 3, wherein the conjugated polymer transfers energy to the signaling chromophore by a mechanism selected from the group consisting of resonant energy transfer, fluorescence resonance energy transfer (FRET), and quantum charge exchange.

5. The composition of claim 3, wherein the energy transfer mechanism is short range.

6. The composition of claim 1, wherein upon excitation of the conjugated polymer with incident light the signaling chromophore produces a light emission that is 8 times greater than a light emission produced upon direct excitation of the signaling chromophore with incident light.

7. The composition of claim 1, wherein upon excitation of the conjugated polymer with incident light the signaling chromophore produces a light emission that is 25 times greater than a light emission produced upon direct excitation of the signaling chromophore with incident light.

8. The composition of claim 1, wherein each repeat unit of the water soluble conjugated polymer comprises a chromophore.