Functionalized chromophoric polymer dots and bioconjugates thereof
The present invention provides, among other aspects, functionalized chromophoric polymer dots comprising a hydrophobic core and a hydrophilic cap, and bioconjugates thereof. Also provided are improved methods for preparing functionalized chromophoric polymer dots. Methods for in vivo imaging and molecular labeling are also disclosed.
1. A functionalized chromophoric polymer dot (Pdot) comprising:
a core and a cap, wherein said core comprises a chromophoric polymer, and said cap comprises a functionalization agent bearing one or more reactive functional groups, with the proviso that less than 100% of the cap is an organo silicate and the proviso that said cap does not encapsulate said core.
2. The functionalized chromophoric polymer dot (Pdot) of claim 1 , having a hydrophobic core and a hydrophilic cap, the Pdot comprising:
(a) the chromophoric polymer; and
(b) an amphiphilic molecule, having a hydrophobic moiety and a hydrophilic moiety, the hydrophilic moiety attached to the one or more reactive functional groups,
wherein:
the chromophoric polymer is embedded within the hydrophobic core of the Pdot;
a hydrophobic portion of the amphiphilic molecule is embedded within the hydrophobic core of the Pdot;
the hydrophilic cap comprises a hydrophilic portion of the amphiphilic molecule; and
the reactive functional group is located in the hydrophilic cap.
3. The functionalized chromophoric polymer dot of claim 1 , wherein the polymer dot comprises a blend of chromophoric polymers.
4. The functionalized chromophoric polymer dot of claim 1 , wherein the chromophoric polymer is a semiconducting polymer.
5. The functionalized chromophoric polymer dot of claim 4 , wherein the polymer dot comprises a blend of semiconducting polymers.
6. The functionalized chromophoric polymer dot of claim 5 , wherein the blend of semiconducting polymers includes a fluorene-benzothiadiazole-thiophen semiconducting polymer.
7. The functionalized chromophoric polymer dot of claim 2 , wherein the weight ratio of the amphiphilic molecule to chromophoric polymer is from about 1% to about 50%.
8. The functionalized chromophoric polymer dot of claim 7 , wherein the weight ratio of the amphiphilic molecule to chromophoric polymer is from about 5% to about 25%.
9. The functionalized chromophoric polymer dot of claim 2 , wherein the hydrophilic moiety of the amphiphilic molecule comprises a polyalkylene glycol.
10. The functionalized chromophoric polymer dot of claim 9 , wherein the polyalkylene glycol is a polyethylene glycol.
11. The functionalized chromophoric polymer dot of claim 2 , wherein the amphiphilic molecule is an amphiphilic polymer.
12. The functionalized chromophoric polymer dot of claim 11 , wherein the amphiphilic polymer is an amphiphilic, comb-like polymer.
13. The functionalized chromophoric polymer dot of claim 11 , wherein the amphiphilic polymer is a polyethylene glycol-grafted polystyrene.
14. The functionalized chromophoric polymer dot of claim 13 , wherein the polyethylene glycol moiety of the amphiphilic polymer is attached to one or more carboxyl groups.
15. The functionalized chromophoric polymer dot of claim 11 , wherein the amphiphilic polymer is a poly(styrene-co-maleic anhydride).
16. The functionalized chromophoric polymer dot of claim 11 , wherein the amphiphilic polymer is a semiconducting polymer.
17. The functionalized chromophoric polymer dot of claim 11 , wherein the amphiphilic polymer is a non-semiconducting polymer.
18. The functionalized chromophoric polymer dot of claim 2 , wherein the hydrophilic moiety of the amphiphilic polymer is attached to more than one reactive functional group.
19. The functionalized chromophoric polymer dot of claim 1 , wherein the one or more reactive functional group is selected from the group consisting of a carboxyl, amino, mercapto, azido, alkyne, aldehyde, hydroxyl, carbonyl, sulfate, sulfonate, phosphate, cyanate, succinimidyl ester, strained alkyne, azide, diene, alkene, tetrazine, strained alkene, cyclooctyne, phosphine groups, and any derivative thereof.
20. The functionalized chromophoric polymer dot of claim 19 , wherein the one or more reactive functional group is an alkyne, azido, strained alkyne, diene, alkene, tetrazine, strained alkene, cyclooctyne, phosphine or other moiety capable of being conjugated to a molecule via a bioorthogonal reaction.
21. The functionalized chromophoric polymer dot of claim 20 , wherein the bioorthogonal reaction is a click chemistry reaction.
22. The functionalized chromophoric polymer dot of claim 1 , wherein a biological molecule is conjugated to a reactive functional group on the surface of the polymer dot.
23. The functionalized chromophoric polymer dot of claim 22 , wherein the biological molecule comprises a molecule selected from the group consisting of a synthetic or naturally occurring protein, a glycoprotein, a polypeptide, an amino acid, a nucleic acid, a carbohydrate, a lipid, a fatty acid, and any combination thereof.
24. The functionalized chromophoric polymer dot of claim 23 , wherein the biological molecule is a polypeptide, an antibody, an aptamer, or a nucleic acid.
25. The functionalized chromophoric polymer dot of claim 1 , wherein the polymer dot is further conjugated to one or more non-reactive chemical groups.
26. The functionalized chromophoric polymer dot of claim 25 , wherein the non-reactive chemical group is a water soluble polymer.
27. The functionalized chromophoric polymer dot of claim 26 , wherein the water soluble polymer is polyethylene glycol.
28. The functionalized chromophoric polymer dot of claim 1 , further comprising an optically inactive polymer chemically cross-linked to the core.
29. The functionalized chromophoric polymer dot of claim 1 , further comprising a second chromophoric polymer chemically cross-linked to the core.
30. The functionalized chromophoric polymer dot of claim 28 , comprising an optically inactive polymer chemically cross-linked to the chromophoric polymer, wherein the chromophoric polymer is a semiconducting polymer.
31. The functionalized chromophoric polymer dot of claim 28 , wherein the optically inactive polymer is an amphiphilic polymer.
32. The functionalized chromophoric polymer dot of claim 28 , wherein the optically inactive polymer comprises a functional group suitable for conjugation to a biological molecule.
33. The functionalized chromophoric polymer dot of claim 32 , further comprising a biological molecule conjugated to the chromophoric polymer dot, wherein:
the biological molecule comprises an amine group;
the functional group is a carboxyl group; and
the amine group is cross-linked to the carboxyl group.
34. The functionalized chromophoric polymer dot of claim 31 , wherein the amphiphilic polymer is a semiconducting polymer.
35. The functionalized chromophoric polymer dot of claim 31 , wherein the amphiphilic polymer comprises a hydrophilic moiety, and wherein the hydrophilic moiety of the amphiphilic polymer is attached to more than one reactive functional group.
36. A functionalized chromophoric polymer dot (Pdot) comprising:
a hydrophobic core comprising a chromophoric polymer; and
a hydrophilic cap, comprising a functionalization agent bearing one or more reactive functional groups,
wherein at least a portion of an exterior surface of the functionalized chromophoric polymer dot is a surface of said hydrophobic core, with the proviso that less than 100% of the cap is an organo silicate.
37. The functionalized chromophoric polymer dot (Pdot) of claim 36 , the Pdot comprising an amphiphilic molecule having a hydrophobic moiety and a hydrophilic moiety, the hydrophilic moiety attached to the one or more reactive functional groups, wherein:
a hydrophobic portion of the amphiphilic molecule is embedded within the hydrophobic core of the Pdot;
the hydrophilic cap comprises a hydrophilic portion of the amphiphilic molecule; and
the one or more reactive functional groups are located in the hydrophilic cap.
38. The functionalized chromophoric polymer dot of claim 36 , wherein the polymer dot comprises a blend of chromophoric polymers.
39. The functionalized chromophoric polymer dot of claim 36 , wherein the chromophoric polymer is a semiconducting polymer.
40. The functionalized chromophoric polymer dot of claim 39 , wherein the polymer dot comprises a blend of semiconducting polymers.
41. The functionalized chromophoric polymer dot of claim 40 , wherein the blend of semiconducting polymers includes a fluorene-benzothiadiazole-thiophen semiconducting polymer.
42. The functionalized chromophoric polymer dot of claim 37 , wherein the weight ratio of the amphiphilic molecule to chromophoric polymer is from about 1% to about 50%.
43. The functionalized chromophoric polymer dot of claim 42 , wherein the weight ratio of the amphiphilic molecule to chromophoric polymer is from about 5% to about 25%.
44. The functionalized chromophoric polymer dot of claim 37 , wherein the hydrophilic moiety of the amphiphilic molecule comprises a polyalkylene glycol.
45. The functionalized chromophoric polymer dot of claim 44 , wherein the polyalkylene glycol is a polyethylene glycol.
46. The functionalized chromophoric polymer dot of claim 37 , wherein the amphiphilic molecule is an amphiphilic polymer.
47. The functionalized chromophoric polymer dot of claim 46 , wherein the amphiphilic polymer is an amphiphilic, comb-like polymer.
48. The functionalized chromophoric polymer dot of claim 46 , wherein the amphiphilic polymer is a polyethylene glycol-grafted polystyrene.
49. The functionalized chromophoric polymer dot of claim 48 , wherein the polyethylene glycol moiety of the amphiphilic polymer is attached to one or more carboxyl groups.
50. The functionalized chromophoric polymer dot of claim 46 , wherein the amphiphilic polymer is a poly(styrene-co-maleic anhydride).
51. The functionalized chromophoric polymer dot of claim 46 , wherein the amphiphilic polymer is a semiconducting polymer.
52. The functionalized chromophoric polymer dot of claim 46 , wherein the amphiphilic polymer is a non-semiconducting polymer.
53. The functionalized chromophoric polymer dot of claim 37 , wherein the hydrophilic moiety of the amphiphilic polymer is attached to more than one reactive functional group.
54. The functionalized chromophoric polymer dot of claim 36 , wherein the one or more reactive functional group is selected from the group consisting of a carboxyl, amino, mercapto, azido, alkyne, aldehyde, hydroxyl, carbonyl, sulfate, sulfonate, phosphate, cyanate, succinimidyl ester, strained alkyne, azide, diene, alkene, tetrazine, strained alkene, cyclooctyne, phosphine groups, and any derivative thereof.
55. The functionalized chromophoric polymer dot of claim 54 , wherein the one or more reactive functional group is an alkyne, azido, strained alkyne, diene, alkene, tetrazine, strained alkene, cyclooctyne, phosphine or other moiety capable of being conjugated to a molecule via a bioorthogonal reaction.
56. The functionalized chromophoric polymer dot of claim 55 , wherein the bioorthogonal reaction is a click chemistry reaction.
57. The functionalized chromophoric polymer dot of claim 36 , wherein a biological molecule is conjugated to a reactive functional group on the surface of the polymer dot.
58. The functionalized chromophoric polymer dot of claim 57 , wherein the biological molecule comprises a molecule selected from the group consisting of a synthetic or naturally occurring protein, a glycoprotein, a polypeptide, an amino acid, a nucleic acid, a carbohydrate, a lipid, a fatty acid, and any combination thereof.
59. The functionalized chromophoric polymer dot of claim 58 , wherein the biological molecule is a polypeptide, an antibody, an aptamer, or a nucleic acid.
60. The functionalized chromophoric polymer dot of claim 36 , wherein the polymer dot is further conjugated to one or more non-reactive chemical groups.
61. The functionalized chromophoric polymer dot of claim 60 , wherein the non-reactive chemical group is a water soluble polymer.
62. The functionalized chromophoric polymer dot of claim 61 , wherein the water soluble polymer is polyethylene glycol.
63. The functionalized chromophoric polymer dot of claim 36 , further comprising an optically inactive polymer chemically cross-linked to the core.
64. The functionalized chromophoric polymer dot of claim 36 , further comprising a second chromophoric polymer chemically cross-linked to the core.
65. The functionalized chromophoric polymer dot of claim 63 , comprising an optically inactive polymer chemically cross-linked to the chromophoric polymer, wherein the chromophoric polymer is a semiconducting polymer.
66. The functionalized chromophoric polymer dot of claim 63 , wherein the optically inactive polymer is an amphiphilic polymer.
67. The functionalized chromophoric polymer dot of claim 63 , wherein the optically inactive polymer comprises a functional group suitable for conjugation to a biological molecule.
68. The functionalized chromophoric polymer dot of claim 67 , further comprising a biological molecule conjugated to the chromophoric polymer dot, wherein:
the biological molecule comprises an amine group;
the functional group is a carboxyl group; and
the amine group is cross-linked to the carboxyl group.
69. The functionalized chromophoric polymer dot of claim 66 , wherein the amphiphilic polymer is a semiconducting polymer.
70. The functionalized chromophoric polymer dot of claim 66 , wherein the amphiphilic polymer comprises a hydrophilic moiety, and wherein the hydrophilic moiety of the amphiphilic polymer is attached to more than one reactive functional group.