Polymers for analyte detection

• US 8,617,819 B2
• Filed: 09/19/2005
• Issued: 12/31/2013
• Est. Priority Date: 09/17/2004
• Status: Active Grant

• Current Assignee: Massachusetts Institute of Technology
• Original Assignee: Massachusetts Institute of Technology
• Inventors: Swager, Timothy M., Seeberger, Peter H., Zheng, Juan, Disney, Matthew D.
• Primary Examiner(s): Haq, Shafiqul
• Application Number:
  US11/230,308
• Publication Number:
  US 20060127929A1
• Time in Patent Office: 3,025 Days
• Field of Search: 436/164, 436/171, 436/172, 422/82.05, 422/82.07, 422/82.08
• US Class Current: 435/7.1
• CPC Class Codes: C12Q 1/6818 involving interaction of two or more labels, e.g. resonant energy transfer
• CPC Class Codes: G01N 2400/10 Polysaccharides, i.e. having more than five saccharide radicals attached to each other by glycosidic linkages; Derivatives thereof, e.g. ethers, esters
• CPC Class Codes: G01N 33/542 with steric inhibition or signal modification, e.g. fluorescent quenching

Abstract

The present invention generally relates to organic polymers able to participate in an analyte-recognition process, where an analyte facilitates an energy transfer between an energy donor and an energy acceptor. Certain embodiments of the invention make use of fluorescent conjugated polymers, such as poly(phenylene ethynylene)s and other polymers comprising pi-conjugated backbones. For example, one aspect of the invention provides a fluorescent conjugated polymer and an indicator that can interact with each other in the presence of an analyte to produce an emissive signal. In some cases, the interaction may include energy exchange mechanisms, such as Dexter energy transfer or the strong coupling effect. The interaction of the conjugated polymer and the indicator, in some instances, may be facilitated through specific interactions, such as a protein/carbohydrate interaction, a ligand/receptor interaction, etc. Another aspect of the invention provides for the detection of biological entities, for example, pathogenic bacteria such as E. coli, or viruses such as influenza virus. In some cases, biological recognition elements may be used to determine the biological entity, for instance, carbohydrates that can be used to specifically interact with at least part of the biological entity, such as a protein in the cell membrane of a bacterium. Still other aspects of the invention involve articles, devices, and kits using any of the above-described systems.

Claims

1. A method of determining a biological entity, comprising:

exposing a sample suspected of containing a biological entity to at least two fluorescent pi-conjugated polymers, each polymer comprising a phenylene ethynylene backbone having at least two phenylene ethynylene groups and a plurality of biological recognition elements covalently attached to the pi-conjugated polymer via the phenyl groups of the phenylene ethynylene backbone, wherein the biological recognition elements are able to specifically interact with the biological entity;

allowing the biological entity, if present, to interact with the biological recognition elements of the at least two fluorescent pi-conjugated polymers in a manner bringing said at least two fluorescent pi-conjugated polymer into proximity with each other such that an emissive signal is produced at a threshold level;

and determining fluorescence of the sample.

2. The method of claim 1, wherein the at least two fluorescent pi-conjugated polymers have the same chemical structure.

3. The method of claim 1, wherein the biological recognition elements are carbohydrates, glycosaminoglycans, glycolipids, proteins, antibodies, glycoproteins, lectins, or nucleic acids.

4. The method of claim 3, wherein the carbohydrates are monosaccharides, oligosaccharides, or polysaccharides.

5. The method of claim 3, wherein the carbohydrates are mannose, fucose, sialic acid, heparin, or the Lewis group antigens.

6. The method of claim 1, wherein the biological entity is a bacterium, a virus, or a protein.

7. The method of claim 1, wherein the biological entity is a cell surface receptor or an enzyme.

8. The method of claim 1, wherein the biological entity is a Escherichia coli, Salmonella entrica, Psuedomonas aerginosa, influenza virus, herpes simplex virus, or Helicobacter pylori.

9. The method of claim 1, wherein the pi-conjugated polymer comprises the structure,

10. The method of claim 9, wherein:

A is a phenyl group;

C is a phenyl group, a triptycene group, or a pentiptycene group;

R is a carbohydrate; and

L is a moiety comprising —

CO—

NH—

, —

(CH₂)_p—

, wherein p is at least 1, —

NH—

CH₂—

CH₂—

, —

CH₂—

CH₂—

CO—

, —

CH₂—

CH₂—

CO—

NH—

CH₂—

CH₂—

, —

O—

CH₂—

CO—

NH—

, —

CH₂—

CH₂—

NH—

, or

11. The method of claim 1, wherein the pi-conjugated polymer comprises the structure,

12. The method as in claim 11, wherein the biological recognition element is mannose.

13. The method as in claim 11, wherein the biological recognition element is galactose.