Polymers for analyte detection

• US 20060127929A1
• Filed: 09/19/2005
• Published: 06/15/2006
• 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., Disney, Matthew D., Seeberger, Peter H., Zheng, Juan
• Granted Patent: US 8,617,819 B2
• Time in Patent Office: Days
• US Class Current: 435/6
• 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 an analyte, comprising:

exposing a sample suspected of containing an analyte to a polymer and an indicator,

wherein, if the sample contains the analyte, the analyte binds both the polymer and indicator in a manner bringing them into proximity with each other such that an emissive signal is produced at a threshold level,

the polymer and the indicator being arranged with respect to each other, prior to the exposing, such that, in the absence of the analyte, the signal at the threshold level does not occur,

the polymer being an energy donor and the indicator being an energy acceptor, each of the polymer and the indicator having a maximum emission wavelength,

wherein the maximum emission wavelength of the polymer and the maximum emission wavelength of the indicator are separated by at least about 100 nm.

2. The method of claim 1, wherein the polymer is a conjugated polymer.

3. The method of claim 1, wherein the polymer is fluorescent.

4. The method of claim 1, wherein the polymer comprises a phenylene ethynylene moiety.

5. The method of claim 1, wherein the polymer comprises a biological recognition element.

6. The method of claim 5, wherein the biological recognition element includes a carbohydrate.

7. The method of claim 5, wherein the biological recognition element includes a saccharide.

29. The method of claim 1, the polymer having a first emission spectrum and the indicator having a second emission spectrum, wherein the first emission spectrum and the second emission spectrum overlap by less than about 10%.

30. The method of claim 29, wherein the overlap is less than about 5%.

31. The method of claim 1, the polymer having a first emission lifetime, the indicator having a second emission lifetime, wherein the first emission lifetime is at least about 100 times greater than the second emission lifetime.

32. The method of claim 1, wherein the polymer has a maximum emission wavelength less than about 700 nanometers.

33. The method of claim 1, wherein the indicator has a maximum emission wavelength greater than about 700 nanometers.

34. The method of claim 1, wherein the polymer and indicator are brought into proximity with each other via a ligand/receptor interaction.

35. The method of claim 1, wherein the polymer is exposed to the indicator prior to or simultaneously with exposure of the polymer to the analyte.

36. The method of claim 1, wherein the analyte is a biological entity.

37. The method of claim 1, wherein the analyte is a biological entity capable of reproduction.

8-28. -28. (canceled)

38. A method of determining an analyte, comprising:

exposing a sample suspected of containing an analyte to a polymer and an indicator,

wherein, if the sample contains the analyte, the analyte binds both the polymer and indicator in a manner bringing them into proximity with each other such that an emissive signal is produced at a threshold level,

the polymer and indicator being arranged with respect to each other, prior to the exposing, such that, in the absence of the analyte, the signal at the threshold level does not occur,

wherein the polymer and indicator are brought into proximity with each other via a ligand/receptor interaction.

39-75. -75. (canceled)

76. A method of determining an analyte, comprising:

exposing a sample suspected of containing an analyte to a polymer and an indicator,

wherein, if the sample contains the analyte, the analyte binds both the polymer and indicator in a manner bringing them into proximity with each other such that an emissive signal is produced at a threshold level,

the polymer and indicator being arranged with respect to each other, prior to the exposing, such that, in the absence of the analyte, the signal at the threshold level does not occur,

wherein the polymer is exposed to the indicator prior to or simultaneously with exposure of the polymer to the analyte.

77-113. -113. (canceled)

114. A method of determining an analyte, comprising:

exposing a sample suspected of containing an analyte to a polymer and an indicator,

wherein, if the sample contains the analyte, the analyte binds both the polymer and indicator in a manner bringing them into proximity with each other such that an emissive signal is produced at a threshold level,

the polymer and indicator being arranged with respect to each other, prior to the exposing, such that, in the absence of the analyte, the signal at the threshold level does not occur,

wherein the analyte is a biological entity capable of reproduction.

115-150. -150. (canceled)

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

exposing a sample suspected of containing a biological entity to a fluorescent conjugated polymer comprising a plurality of biological recognition elements, at least some of which are able to specifically interact with the biological entity; and

determining fluorescence of the sample.

152-185. -185. (canceled)