General signaling protocol for chemical receptors in immobilized matrices

• US 6,589,779 B1
• Filed: 07/14/2000
• Issued: 07/08/2003
• Est. Priority Date: 07/16/1999
• Status: Expired due to Term

• Current Assignee: University of Texas System
• Original Assignee: Board of Regents of the University of Texas System (University of Texas System)
• Inventors: Neikirk, Dean P., Anslyn, Eric V., McDevitt, John T., Shear, Jason B.
• Primary Examiner(s): Redding, David A.
• Application Number:
  US09/616,482
• Time in Patent Office: 1,089 Days
• Field of Search: 435/287.1, 435/287.2, 435/288.7
• US Class Current: 435/288.7
• CPC Class Codes: B01L 2200/0668 Trapping microscopic beads
• CPC Class Codes: B01L 2300/0636 Integrated biosensor, microarrays
• CPC Class Codes: B01L 2300/0819 Microarrays; Biochips
• CPC Class Codes: B01L 2300/0877 Flow chambers
• CPC Class Codes: B01L 2400/049 vacuum
• CPC Class Codes: B01L 3/502761 specially adapted for handling suspended solids or molecules independently from the bulk fluid flow, e.g. for trapping or sorting beads, for physically stretching molecules investigating characteristics of particles G01N15/00
• CPC Class Codes: G01N 2021/7786 Fluorescence
• CPC Class Codes: G01N 21/253 for batch operation, i.e. multisample apparatus analytical automats G01N35/00
• CPC Class Codes: G01N 21/6452 Individual samples arranged in a regular 2D-array, e.g. multiwell plates
• CPC Class Codes: G01N 21/77 by observing the effect on a chemical indicator
• CPC Class Codes: G01N 21/78 producing a change of colour

Abstract

A system for the rapid characterization of multi-analyte fluids, in one embodiment, includes a light source, a sensor array, and a detector. The sensor array is formed from a supporting member into which a plurality of cavities may be formed. A series of chemically sensitive particles are, in one embodiment positioned within the cavities. The particles may be configured to produce a signal when a receptor coupled to the particle interacts with the analyte. Using pattern recognition techniques, the analytes within a multi-analyte fluid may be characterized.

Claims

1. A system for detecting an analyte in a fluid comprising:

a light source;

a sensor array, the sensor array comprising;

a supporting member comprising at least one cavity formed within the supporting member; and

a top cover layer, wherein the top cover layer is coupled to a top surface of the supporting member such that a particle is contained within the cavity by the top cover layer;

the particle positioned within the cavity, wherein the particle comprises a receptor coupled to a polymeric resin, and an indicator coupled to the polymeric resin, and wherein the indicator is configured to produce a signal when the receptor interacts with the analyte during use;

a detector, the detector being configured to detect the signal produced by the interaction of the analyte with the particle during use;

wherein the light source and detector are positioned such that light passes from the light source, to the particle, and onto the detector during use.

2. The system of claim 1, wherein the system further comprises a plurality of particles positioned within a plurality of cavities, and wherein the system is configured to substantially simultaneously detect a plurality of analytes in the fluid.

3. The system of claim 1, wherein the system further comprises a plurality of particles positioned within the cavity.

4. The system of claim 1, wherein the light source comprises a light emitting diode.

5. The system of claim 1, wherein the light source comprises a white light source.

6. The system of claim 1, wherein the sensor array further comprises a bottom layer, wherein the bottom layer is positioned below a bottom surface of the supporting member, and wherein the bottom layer is positioned such that the particle is substantially contained within the cavity by the bottom layer and the top cover layer.

7. The system of claim 6, wherein the bottom layer and the top cover layer are substantially transparent to light produced by the light source.

8. The system of claim 1, wherein the sensor array further comprises a bottom layer, wherein the bottom layer is coupled to a bottom surface of the supporting member, and wherein both the bottom layer is coupled to the supporting member such that the particle is substantially contained within the cavity by bottom layer and the top cover layer.

9. The system of claim 8, wherein the bottom layer and the top cover layer are substantially transparent to light produced by the light source.

10. The system of claim 1, wherein the sensor array further comprises a bottom layer coupled to the supporting member, and wherein the supporting member comprises silicon, and wherein the bottom layer comprises silicon nitride.

11. The system of claim 1, wherein the sensor array further comprises a sensing cavity formed on a bottom surface of the sensor array.

12. The system of claim 1, wherein the supporting member is formed from a plastic material, and wherein the top cover layer is configured to allow the fluid to pass through the top cover layer to the particle, and wherein both the supporting member and the top cover layer are substantially transparent to light produced by the light source.

13. The system of claim 1, further comprising a fluid delivery system coupled to the supporting member.

14. The system of claim 1, wherein the detector comprises a charge-coupled device.

15. The system of claim 1, wherein the detector comprises an ultraviolet detector.

16. The system of claim 1, wherein the detector comprises a fluorescence detector.

17. The system of claim 1, wherein the detector comprises a semiconductor based photodetector, and wherein the detector is coupled to the sensor array.

18. The system of claim 1, wherein the particle ranges from about 0.05 micron to about 500 microns.

19. The system of claim 1, wherein a volume of the particle changes when contacted with the fluid.

20. The system of claim 1, wherein the polymeric resin comprises polystyrene-polyethylene glycol-divinyl benzene.

21. The system of claim 1, wherein the receptor comprises a polynucleotide.

22. The system of claim 1, wherein the receptor comprises a peptide.

23. The system of claim 1, wherein the receptor comprises a compound of the general formula:

24. The system of claim 1, wherein the receptor comprises a compound of the general formula:

 

(R¹)_n—

X—

(R²)_m

wherein X comprises carbocyclic systems or C₁-C₁₀ alkanes, n is an integer of at least 1, m is an integer of at least 1; and

wherein each of R¹ independently represents —

(CH₂)_y—

NH—

Y;

where y is an integer of at least 1;

where R³, R⁴, and R⁵ independently represent hydrogen, alkyl, aryl, alkyl carbonyl of 1 to 10 carbon atoms, or alkoxy carbonyl of 1 to 10 carbon atoms, or R⁴ and R⁵ together represent a cycloalkyl group;

where R⁶ represents hydrogen, alkyl, aryl, alkyl carbonyl of 1 to 10 carbon atoms, or alkoxy carbonyl of 1 to 10 carbon atoms;

where R⁷ represents alkyd, aryl, alkyl carbonyl of 1 to 10 carbon atoms, or alkoxy carbonyl of 1 to 10 carbon atoms;

where R⁶ and R⁷ together represent a cycloalkyl group;

where Y is a peptide or hydrogen; and

wherein each of R² independently represents hydrogen, alkyl, alkenyl, alkynyl, phenyl, phenylalkyl, arylalkyl, aryl, or together with another R² group represent a carbocyclic ring.

25. The system of claim 1, wherein the receptor comprises a compound of the general formula:

26. The system of claim 1, wherein the receptor comprises a compound of the general formula:

27. The system of claim 1, wherein the receptor comprises an enzyme.

28. The system of claim 1, wherein the receptor comprises a peptide mimetic.

29. The system of claim 1, wherein the receptor is coupled to the polymeric resin by a linker.

30. The system of claim 1, wherein the receptor is coupled to the polymeric resin by a first linker and wherein the indicator is coupled to the polymeric resin by a second linker.

31. The system of claim 1, wherein the receptor is coupled to the polymeric resin by a first linker and wherein the indicator is coupled to the polymeric resin by a second linker, and wherein the particle further comprises an additional indicator coupled to the polymeric resin by a third linker, wherein the interaction of the receptor with the analyte causes the indicator and the additional indicator to interact such that the signal is produced.

32. The system of claim 1, wherein the receptor is coupled to the polymeric resin by a first linker and wherein the indicator is coupled to the receptor.

33. The system of claim 1, wherein the receptor is coupled to the polymeric resin by a first linker and wherein the indicator is coupled to the receptor, and wherein the particle further comprises an additional indicator coupled to the receptor, wherein the interaction of the receptor with the analyte causes the indicator and the additional indicator to interact such that the signal is produced.

34. The system of claim 1, wherein the receptor is coupled to the polymeric resin by a first, linker and wherein the indicatoy is coupled to the receptor by a second linker.

35. The system of claim 1, wherein the receptor is coupled to the polymeric resin by a first linker and wherein the indicator is coupled to the receptor by a second linker, and wherein the particle further comprises an additional indicator coupled to the receptor, wherein the interaction of the receptor with the analyte causes the indicator and the additional indicator to interact such that the signal is produced.

36. The system of claim 1, wherein the receptor is coupled to the polymeric resin by a first linker and wherein the indicator is, coupled to the first linker.

37. The system of claim 1, wherein the receptor is coupled to the polymeric resin by a first linker, and wherein the indicator is coupled to the first linker by a second linker.

38. The system of claim 1, wherein the receptor is coupled to the polymeric resin by a first linker, and wherein the indicator is coupled to the first linker by a second linker, and wherein the particle further comprises an additional indicator coupled to the receptor, wherein the interaction of the receptor with the analyte causes the indicator and the additional indicator to interact such that the signal is produced.

39. The system of claim 1, wherein the receptor is coupled to the polymeric resin by a first linker, and wherein the indicator is coupled to the first linker by a second linker, and wherein the particle further comprises an additional indicator coupled to the first linker by a third linker, wherein the interaction of the receptor with the analyte causes the indicator and the additional indicator to interact such that the signal is produced.

40. The system of claim 1, wherein the indicator interacts with the receptor in the absence of an analyte.

41. The system of claim 1, wherein the particle further comprises an additional indicator coupled to the polymeric resin, and wherein the indicator is a first fluorescent dye and wherein the additional indicator is a second fluorescent dye, and wherein the indicator and the additional indicator produce a fluorescence resonance energy transfer signal, and wherein the interaction of the analyte with the receptor causes the distance between the indicator and the additional indicator to become altered such that the fluorescence resonance energy transfer signal is altered.

42. The system of claim 1, wherein the particle further comprises an additional indicator coupled to the polymeric resin, wherein the indicator is a fluorescent dye and wherein the additional indicator is a fluorescence quencher, and wherein the indicator and the additional indicator are positioned such that the fluorescence of the indicator is at least partially quenched by the additional indicator, and wherein the interaction of the analyte with the receptor causes the distance between the indicator and the additional indicator to become altered such that the absorbance of the fluorescence of the indicator by the additional indicator is altered.

43. The system of claim 1, wherein the particle ranges from about 0.05 micron to about 500 microns.

44. The system of claim 1, wherein the polymeric resin comprises polystyrene-polyethylene glycol-divinyl benzene.

45. The system of claim 1, wherein the system further comprises a plurality of particles positioned within a plurality of cavities, and wherein the plurality of particles produce a detectable pattern in the presence of the analyte.