Conjugated polymer tag complexes

• US 8,048,659 B1
• Filed: 10/07/2000
• Issued: 11/01/2011
• Est. Priority Date: 10/08/1999
• Status: Expired due to Fees

• Current Assignee: Lidia Vallarino, Robert C. Leif
• Original Assignee: Lidia Vallarino, Robert C. Leif
• Inventors: Vallarino, Lidia, Leif, Robert C.
• Primary Examiner(s): COOK, LISA V
• Application Number:
  US10/110,114
• Time in Patent Office: 4,042 Days
• Field of Search: 435/174, 436/518, 422/55, 424/9.322, 424/78.08
• US Class Current: 435/174
• CPC Class Codes: C08F 289/00 Macromolecular compounds obtained by polymerising monomers on to macromolecular compounds not provided for in groups C08F251/00 - C08F287/00
• CPC Class Codes: C08F 290/14 Polymers provided for in subclass C08G
• CPC Class Codes: C08F 291/00 Macromolecular compounds obtained by polymerising monomers on to macromolecular compounds according to more than one of the groups C08F251/00 - C08F289/00
• CPC Class Codes: C08L 51/08 grafted on to macromolecular compounds obtained otherwise than by reactions only involving unsaturated carbon-to-carbon bonds
• CPC Class Codes: C12Q 1/37 involving peptidase or proteinase
• CPC Class Codes: G01N 33/582 with fluorescent label

Abstract

A tagged water-soluble polymer linked to a solid support is provided and includes a solid support optionally including a spacer sequence including one or more spacer monomer units attached to the solid support; and a water-soluble polymer covalently linked to the solid support or to the spacer sequence, the water-soluble polymer including a first cleavage segment including at least one monomer unit including a selectively cleavable link to the solid support or to the spacer sequence, where cleavage of the selectively cleavable link separates the water-soluble polymer from the solid support, a second segment including at least one monomer unit selected from a monomer unit linked to a reactive functionality able to be covalently coupled to a tag or linked to a tag, a third segment including one or more monomer units linked to a reactive functionality that can form a covalent bond with an analyte-binding species or an analyte, and optionally one or more spacer monomer units provided adjacent to any segment and/or between any two segments and/or within any one or more of the second segment and the third segment.

Claims

1. A water-soluble polymer linked to a solid support, comprising:

a solid support optionally comprising a spacer sequence comprising one or more spacer monomer units attached to the solid support; and

a water-soluble polymer covalently linked to the solid support or to the spacer sequence, the water-soluble polymer comprising

a first cleavage segment of known composition and/or sequence comprising at least one monomer unit comprising a selectively cleavable link to the solid support or to the spacer sequence, wherein cleavage of the selectively cleavable link separates the water-soluble polymer from the solid support,

a second segment of known composition and/or sequence comprising at least one monomer unit selected from the group consisting of a monomer unit linked to a reactive functionality able to be covalently coupled to a tag and a monomer unit linked to a tag,

a third segment of known composition and/or sequence comprising one or more monomer units linked to a reactive functionality that can form a covalent bond with a member of a specific combining pair, and

optionally one or more spacer monomer units provided adjacent to any segment and/or between any two segments and/or within any one or more of the second segment and the third segment.

2. The water-soluble polymer linked to a solid support of claim 1, wherein the second segment of known composition and/or sequence comprises at least one monomer unit selected from the group consisting of a monomer unit linked to a reactive functionality and a monomer unit linked to a tag.

3. The water-soluble polymer linked to a solid support of claim 2, wherein the cleavage of the selectively cleavable link does not decompose the tag.

4. The water-soluble polymer linked to a solid support of claim 1, wherein the third segment of known composition and/or sequence comprises one or more monomer units linked to a reactive functionality covalently bound to a member of a specific combining pair.

5. The water-soluble polymer linked to a solid support of claim 4, wherein the cleavage of the selectively cleavable link does not severe the linkage of the covalently bound member of a specific combining pair from the polymer or interfere with the formation of the combining pair.

6. The water-soluble polymer linked to a solid support of claim 1, wherein the solid support is a water-insoluble, swellable functionalized bead having a dry particle size in the range from 10 to 500 microns and at least one of the monomers of the polymer includes a functionalized hydrophobic side chain.

7. The water-soluble polymer linked to a solid support of claim 6, wherein the solid support comprises a spacer sequence comprising one or more spacer monomer units attached to the solid support.

8. The water-soluble polymer linked to a solid support of claim 2, wherein the reactive functionalities of the water-soluble polymer are independently selected from the group consisting of amino, azide, alcoholic hydroxyl, phenolic hydroxyl, aldehyde, carboxylic acid, carboxamide, halogen, isocyanate, isothiocyanate, mercapto, activated disulfide, nitrile, functionalized alkyl, functionalized aryl, and functionalized alkyl-substituted aryl substituents.

9. The water-soluble polymer linked to a solid support of claim 1, wherein the third segment comprises at least one monomer unit having a reactive functionality linked to an analyte-binding species or an analyte.

10. The water-soluble polymer linked to a solid support of claim 4, wherein the member of a specific combining pair able to form a covalent bond with the reactive functionality of the third segment is selected from the group consisting of an analyte-binding species or an analyte.

11. The water-soluble polymer linked to a solid support of claim 1, wherein the water-soluble polymer consists essentially of monomer units able to undergo synthesis on a solid support, the monomer units being selected from the group consisting of amino acids, peptide nucleic acids, nucleotide monophosphate monomer units, and sugar monomer units.

12. The water-soluble polymer linked to a solid support of claim 1, wherein the first cleavage segment comprises a sequence of at least two linked monomer units, each monomer unit selected from the group consisting of an amino acid monomer, a sugar monomer and a nucleotide, and wherein the water-soluble polymer is selectively cleavable from said support in the presence of an enzyme.

13. The water-soluble polymer linked to a solid support of claim 12, wherein the first cleavage segment comprises at least one disulfide linked monomer units that is selectively cleavable from said support in the presence of a reducing agent.

14. The water-soluble polymer linked to a solid support of claim 12, wherein the cleavage segment is a substrate for the enzyme and the enzyme is selected from the group consisting of a proteinase, an endonuclease and a glycosidase.

15. The water-soluble polymer linked to a solid support of claim 2, wherein at least one tag is selected from the group consisting of an optical-label, an other-label, or a separation-tag.

16. The water-soluble polymer linked to a solid support of claim 15, wherein at least one tag is an optical-label.

17. The water-soluble polymer linked to a solid support of claim 16, wherein the second segment comprises at least a first monomer unit having a first optical-label and a second monomer unit having a second optical-label, wherein the first optical-label is different from the second optical-label.

18. The water-soluble polymer linked to a solid support of claim 17, wherein the first optical-label is a luminescence enhancer and the second optical-label is a lanthanide complex.

19. The water-soluble polymer linked to a solid support of claim 17, wherein the first optical-label is capable of transferring energy to the second optical-label.

20. The water-soluble polymer linked to a solid support of claim 19, wherein the second segment further comprises at least a third monomer having a third optical-label different from the first and the second optical-labels, the optical-labels being ordered into pairs with the emission of a first optical-label overlapping the excitation of the second optical-label, the emission of the second optical-label overlapping the excitation of the third optical-label;

and wherein the ordering of monomer units produces a relative orientation and spacing of the optical-labels to maximize energy transfer between the optical-label with the shortest excitation wavelength and the optical-label with the longest emission wavelength.

21. The water-soluble polymer linked to a solid support of claim 20, wherein each of the first, second, and third optical-labels is an independent species that emits light in a range of from 300 to 1,400 nanometers after receiving energy in a range of from 200 to 1,000 nanometers.

22. The water-soluble polymer linked to a solid support of claim 17, wherein the first optical-label and second optical-label are sequentially ordered to control their interactions.

23. The water-soluble polymer linked to a solid support of claim 11, wherein the monomer units comprise amino acid monomer units linked by amide linkages.

24. The water-soluble polymer linked to a solid support of claim 23, wherein each reactive functionality is independently selected from the group consisting of amino, azide, alcoholic hydroxyl, phenolic hydroxyl, aldehyde, carboxylic acid, carboxamide, halogen, isocyanate, isothiocyanate, mercapto, activated disulfide, nitrile, functionalized alkyl, functionalized aryl, and functionalized alkyl-substituted aryl substituents.

25. The water-soluble polymer linked to a solid support of claim 23, wherein at least one tag is an optical-label.

26. The water-soluble polymer linked to a solid support of claim 25, wherein at least one tag is an optical-label that is capable of absorbing light in a certain wavelength range and emitting light in a higher wavelength range.

27. The water-soluble polymer linked to a solid support of claim 25, wherein at least one amino acid monomer unit absorbs light in a range different from that of the optical-label.

28. The water-soluble polymer linked to a solid support of claim 25, wherein the optical-label is a lanthanide compound.

29. The water-soluble polymer linked to a solid support of claim 17, further comprising at least one spacer monomer unit disposed such that the first optical-label has an orientation and spacing relative to the second optical-label to maximize energy transfer between the first and second optical-labels.

30. The water-soluble polymer linked to a solid support of claim 25, wherein the optical-label is capable of absorbing and/or emitting light in a wavelength range of from 200 to 1,400 nanometer.

31. The water-soluble polymer linked to a solid support of claim 30, wherein the optical-label is capable of emitting light in a range of from 300 to 1,400 nanometers.

32. The water-soluble polymer linked to a solid support of claim 30, wherein the optical-label is capable of absorbing light in a range of from 200 to 1,000 nanometers.

33. The water-soluble polymer linked to a solid support of claim 17, wherein the first optical-label is capable of transferring energy to the second optical-label.

34. The water-soluble polymer linked to a solid support of claim 28, wherein the lanthanide compound is a lanthanide complex.

35. The water-soluble polymer linked to a solid support of claim 34, wherein the lanthanide complex is a lanthanide macrocycle.

36. The water-soluble polymer linked to a solid support of claim 34, wherein the lanthanide complex includes at least one enhancer.

37. The water-soluble polymer linked to a solid support of claim 28, wherein the lanthanide compound is selected from the group consisting of a compound of europium, a compound of samarium, a compound of terbium, and a compound of dysprosium.

38. The water-soluble polymer linked to a solid support of claim 28, wherein the lanthanide compound is an energy transfer acceptor lanthanide macrocycle compound having an excitation spectrum maximum in a range of from 200 to 700 nanometers.

39. The water-soluble polymer linked to a solid support of claim 36, wherein the enhancer of an energy transfer acceptor lanthanide macrocycle complex having an excitation spectrum maximum in a range of from 200 to 700 nanometers.

40. The water-soluble polymer linked to a solid support of claim 28, wherein the lanthanide compound is an energy transfer acceptor lanthanide macrocycle complex having an emission spectrum maximum in a range of from 500 to 1400 nanometers.

41. The water-soluble polymer linked to a solid support of claim 36, wherein the lanthanide compound accepts energy from a luminescence enhancer.

42. The water-soluble polymer linked to a solid support of claim 36, wherein the lanthanide complex accepts energy from a luminescence enhancer.

43. The water-soluble polymer linked to a solid support of claim 41, wherein a second lanthanide ion is involved with the transfer of energy to the lanthanide complex.

44. The water-soluble polymer linked to a solid support of claim 18, wherein the lanthanide complex has an orientation and spacing relative to the luminescence enhancer so as to maximize the emission of light.

45. The water-soluble polymer linked to a solid support of claim 18, wherein the luminescence enhancer is selected from the group consisting of a beta-diketone, a beta-diketonate, and a mixture thereof.

46. The water-soluble polymer linked to a solid support of claim 33, wherein the first optical-label and the second optical-label are an organic optical-label pair.

47. The water-soluble polymer linked to a solid support of claim 46, wherein the second optical-label after receiving energy from the first optical-label, emits light in a range of from 300 to 1,400 nanometers.

48. The water-soluble polymer linked to a solid support of claim 15, wherein the at least one tag is an other-label.

49. The water-soluble polymer linked to a solid support of claim 48, wherein the other-label is radioactive.

50. The water-soluble polymer linked to a solid support of claim 48, wherein the other-label is paramagnetic.

51. The water-soluble polymer linked to a solid support of claim 15, wherein the at least one tag is a separation-tag.

52. The water-soluble polymer linked to a solid support of claim 51, wherein the separation-tag is a moiety that increases magnetic susceptibility, ionic charge, mass, or density.

53. The water-soluble polymer linked to a solid support of claim 15, wherein the second segment comprises at least one tag that is an optical-label and one or more tags independently selected from the group consisting of an other-label and a separation-tag.

54. The water-soluble polymer linked to a solid support of claim 10, wherein the third segment further comprises an analyte-binding species covalently bound to the reactive functionality, and wherein the analyte-binding species is selected from the group consisting of

an antibody,

a protein,

a nucleic acid,

a peptide nucleic acid,

a polysaccharide, and

a haptene having a molecular weight in the range of from 125 to 2,000 daltons.

55. The water-soluble polymer linked to a solid support of claim 54, wherein the antibody is a polyclonal antibody or a monoclonal antibody.

56. The water-soluble polymer linked to a solid support of claim 54, wherein the nucleic acid is DNA or RNA.

57. The water-soluble polymer linked to a solid support of claim 11, wherein the polymer is a dendrimer.

58. The water-soluble polymer linked to a solid support of claim 6, wherein the polymer is a dendrimer.