Multiple-property composite beads and preparation and use thereof

• US 20030178309A1
• Filed: 03/21/2002
• Published: 09/25/2003
• Est. Priority Date: 03/21/2002
• Status: Active Grant

• Current Assignee: AVIVA Biosciences Corporation
• Original Assignee: AVIVA Biosciences Corporation
• Inventors: Wang, Xiaobo, Wu, Lei, Cheng, Jing, Xu, Junquan, Huang, Mingxian, Tao, Guo Liang
• Granted Patent: US 7,214,427 B2
• Time in Patent Office: Days
• US Class Current: 204/547
• CPC Class Codes: B01J 19/0046 Sequential or parallel reactions, e.g. for the synthesis of polypeptides or polynucleotides; Apparatus and devices for combinatorial chemistry or for making molecular arrays synthesis methods per se C40B50/00
• CPC Class Codes: B01J 2219/00459 Beads
• CPC Class Codes: B01J 2219/005 Beads
• CPC Class Codes: B01J 2219/00545 Colours
• CPC Class Codes: B01J 2219/00551 3-dimensional
• CPC Class Codes: B01J 2219/00585 Parallel processes
• CPC Class Codes: B01J 2219/00596 Solid-phase processes
• CPC Class Codes: B01J 2219/00655 the compounds being bound to magnets embedded in or on the solid supports
• CPC Class Codes: B01J 2219/0072 Organic compounds
• CPC Class Codes: B01J 2219/0074 Biological products
• CPC Class Codes: B03C 1/02 acting directly on the substance being separated
• CPC Class Codes: B03C 5/026 using open-gradient differential dielectric separation, i.e. using electrodes of special shapes for non-uniform field creation, e.g. Fluid Integrated Circuit [FIC]
• CPC Class Codes: B03C 5/028 using travelling electric fields, i.e. travelling wave dielectrophoresis [TWD]
• CPC Class Codes: B82Y 15/00 Nanotechnology for interacting, sensing or actuating, e.g. quantum dots as markers in protein assays or molecular motors
• CPC Class Codes: C40B 40/06 Libraries containing nucleotides or polynucleotides, or derivatives thereof
• CPC Class Codes: C40B 50/16 involving encoding steps
• CPC Class Codes: C40B 70/00 Tags or labels specially adapted for combinatorial chemistry or libraries, e.g. fluorescent tags or bar codes
• CPC Class Codes: G01N 33/54326 Magnetic particles
• CPC Class Codes: G01N 33/588 with semiconductor nanocrystal label, e.g. quantum dots
• CPC Class Codes: Y10S 428/90 Magnetic feature
• CPC Class Codes: Y10T 428/2982 : Particulate matter [e.g., sphere, flake, etc.]
• CPC Class Codes: Y10T 428/2991 : Coated
• CPC Class Codes: Y10T 428/32 : Composite [nonstructural laminate] of inorganic material having metal-compound-containing layer and having defined magnetic layer

Abstract

This invention relates generally to the field of moiety or molecule isolation, detection and manipulation and library synthesis. In particular, the invention provides a bead, which bead comprises: a) a magnetizable substance; and b) an electrically conductive substance or an optical labeling substance. Methods and kits for isolating, detecting and manipulating moieties and synthesizing libraries using the beads are also provided.

Claims

1. A bead, which bead comprises:

a) a magnetizable substance; and

b) an electrically conductive substance or an optical labeling substance.

2. The bead of claim 1, wherein the magnetizable substance is selected from the group consisting of a paramagnetic substance, a ferromagnetic substance and a ferrimagentic substance.

3. The bead of claim 1, wherein the magnetizable substance comprises a metal composition.

4. The bead of claim 3, wherein the metal composition is a transition metal composition or an alloy thereof.

5. The bead of claim 4, wherein the transition metal is selected from the group consisting of iron, nickel, copper, cobalt, manganese, tantalum, zirconium and cobalt-tantalum-zirconium (CoTaZr) alloy.

6. The bead of claim 3, wherein the metal composition is Fe₃O₄.

7. The bead of claim 1, wherein the electrically conductive substance is selected from the group consisting of gold, silver, a carbon particle and a conductive polymer.

8. The bead of claim 7, wherein the conductive polymer is a polyaniline, a polypyrrole, or a polythiophene.

9. The bead of claim 1, wherein the electrically conductive substance allows the bead to exhibit positive dielectrophoresis force.

10. The bead of claim 1, wherein the optical labeling substance is selected from the group consisting of a fluorescent substance, a scattered-light detectable particle and a quantum dot.

11. The bead of claim 10, wherein the quantum dot comprises a Cd-X core, X being Se, S or Te.

12. The bead of claim 11, wherein the quantum dot is passivated with an inorganic coating shell.

13. The bead of claim 12, wherein the coating shell comprises Y-Z, Y being Cd or Zn, and Z being S or Se.

14. The bead of claim 10, wherein the quantum dot comprises a Cd-X core, X being Se, S or Te, a Y-Z shell, Y being Cd or Zn, and Z being S or Se, and the bead is further overcoated with a trialkylphosphine oxide.

15. The bead of claim 1, which comprises a plurality of optical labeling substances.

16. The bead of claim 15, which comprises at least two different types of optical labeling substances.

17. The bead of claim 10, which comprises a plurality of quantum dots.

18. The bead of claim 17, wherein at least two quantum dots have different sizes.

19. The bead of claim 10, wherein the quantum dot has a size ranging from about 1 nm to about 100 nm.

20. The bead of claim 1, which comprises a magnetizable substance and an electrically conductive substance.

21. The bead of claim 1, which comprises a magnetizable substance and an optical labeling substance.

22. The bead of claim 1, which further comprises a binding partner that is capable of binding to a moiety.

23. The bead of claim 22, wherein the binding partner specifically binds to the moiety.

24. The bead of claim 22, wherein the binding partner is selected from the group consisting of a cell, a cellular organelle, a virus, a molecule and an aggregate or complex thereof.

25. The bead of claim 1, which comprises a magnetizable substance, an electrically conductive substance and an optical labeling substance.

26. The bead of claim 1, which comprises a magnetizable substance, an electrically conductive substance and a quantum dot.

27. The bead of claim 25, which further comprises a binding partner that is capable of binding to a moiety.

28. The bead of claim 27, wherein the binding partner specifically binds to the moiety.

29. The bead of claim 27, wherein the binding partner is selected from the group consisting of a cell, a cellular organelle, a virus, a molecule and an aggregate or complex thereof.

30. The bead of claim 1, which has a size ranging from about 0.05 μ

m to about 10 μ

m.

31. The bead of claim 1, which has a spherical, a cubical or other regular or irregular shape.

32. The bead of claim 1, which has a magnetizable substance core coated with an electrically conductive substance, a quantum dot and a binding partner that is capable of binding to a moiety.

33. A method for isolating a moiety, which method comprises:

a) providing a bead of claim 1 comprising an optical labeling substance and a binding partner that is capable of binding to a moiety to be isolated;

b) contacting a sample containing or suspected of containing of said moiety with said bead provided in step a) under conditions allowing binding between said moiety and said binding partner; and

c) recovering said bead from said sample,

whereby the identity of said isolated moiety is assessed by analyzing said optical labeling substance comprised in said bead.

34. The method of claim 33, wherein the optical labeling substance is a quantum dot.

35. The method of claim 33, wherein the moiety is a cell, a cellular organelle, a virus, a molecule and an aggregate or complex thereof.

36. The method of claim 33, wherein the bead is recovered from the sample by a magnetic field, centrifugation or filtration.

37. The method of claim 33, wherein a plurality types of moieties are isolated by using a plurality types of beads, each type of the beads contains a binding partner that is capable of binding to a member of the plurality types of the moieties.

38. The method of claim 33, wherein the sample is a fluid sample.

39. The method of claim 33, wherein the isolation is conducted in a liquid container selected from the group consisting of a beaker, a flask, a cylinder, a test tube, an enpindorf tube, a centrifugation tube, a culture dish, a multiwell plate and a filter membrane.

40. The method of claim 33, further comprising a step of recovering said isolated moiety from said bead.

41. The method of claim 33, wherein the binding partner specifically binds to the moiety.

42. A method for manipulating a moiety, which method comprises:

a) providing a bead of claim 1 comprising a binding partner that is capable of binding to a moiety to be manipulated;

b) coupling said moiety to said bead provided in step a) via binding between said moiety and said binding partner to form a moiety-bead complex; and

c) manipulating said moiety-bead complex with a dielectrophoresis, a traveling-wave dielectrophoresis and/or a magnetic force, thereby said moiety is manipulated.

43. The method of claim 42, wherein the moiety-bead complex is manipulated in a chip format and the manipulation is effected through a combination of a structure that is external to the chip and a structure that is built-in in the chip.

44. The method of claim 42, wherein the moiety to be manipulated is selected from the group consisting of a cell, a cellular organelle, a virus, a molecule and an aggregate or complex thereof.

45. The method of claim 42, wherein the manipulation is selected from the group consisting of transportation, focusing, enrichment, concentration, aggregation, trapping, repulsion, levitation, separation, fractionation, isolation and linear or other directed motion of the moiety.

46. The method of claim 42, wherein the moiety is not directly manipulatable by a dielectrophoresis, a traveling-wave dielectrophoresis and/or a magnetic force.

47. The method of claim 42, wherein neither the moiety nor the binding partner is directly manipulatable by a dielectrophoresis, a traveling-wave dielectrophoresis and/or a magnetic force.

48. The method of claim 42, wherein at least two types of moieties are manipulated.

49. The method of claim 48, wherein the moieties are manipulated via a plurality of types of corresponding beads.

50. The method of claim 48, wherein the moieties are manipulated sequentially or simultaneously.

51. The method of claim 42, further comprising a step of recovering said isolated moiety from said bead.

52. The method of claim 42, wherein the bead comprises an optical labeling substance and the method further comprises a step of assessing the identity of the manipulated moiety by analyzing the optical labeling substance comprised in the bead.

53. The method of claim 52, wherein the optical labeling substance is a quantum dot.

54. The method of claim 51, wherein the bead comprises an optical labeling substance and the method further comprises a step of assessing the identity of the isolated moiety by analyzing the optical labeling substance comprised in the bead.

55. The method of claim 54, wherein the optical labeling substance is a quantum dot.

56. The method of claim 42, wherein the binding partner specifically binds to the moiety.

57. A kit for manipulating a moiety, which kit comprises:

a) a bead of claim 1 comprising a binding partner that is capable of binding to a moiety to be manipulated; and

b) a chip on which a moiety-bead complex can be manipulated.

58. The kit of claim 57, further comprising instruction(s) for coupling the moiety to the bead and/or for manipulating the moiety-bead complex on the chip.

59. The kit of claim 57, further comprising a chamber comprising the chip.

60. A method for detecting a moiety, which method comprises:

a) providing a bead of claim 1 comprising an optical labeling substance and a binding partner that is capable of binding to a moiety to be detected;

b) contacting a sample containing or suspected of containing of said moiety with said bead provided in step a) under conditions allowing binding between said moiety and said binding partner; and

c) detecting binding between said moiety and said binding partner,

whereby the presence or amount of said moiety is assessed by analysis of binding between said moiety and said binding partner and the identity of said moiety is assessed by analyzing the optical labeling substance comprised in the bead.

61. The method of claim 60, wherein the optical labeling substance is a quantum dot.

62. The method of claim 60, wherein the moiety is a cell, a cellular organelle, a virus, a molecule and an aggregate or complex thereof.

63. The method of claim 60, wherein a plurality of moieties is detected by using a plurality of beads, each of the beads contains a binding partner that is capable of binding to a member of the plurality of the moieties.

64. The method of claim 63, wherein the presence, amount or identity of said moieties are detected simultaneously.

65. The method of claim 60, wherein the sample is a fluid sample.

66. The method of claim 60, wherein the sample is contacted with the bead in a liquid container selected from the group consisting of a beaker, a flask, a cylinder, a test tube, an enpindorf tube, a centrifugation tube, a culture dish, a multiwell plate and a filter membrane.

67. The method of claim 60, wherein the binding partner specifically binds to the moiety.

68. A method for synthesizing a library, which method comprises:

a) providing a plurality of beads of claim 1, each of said beads comprises an optical labeling substance that corresponds to an entity to be synthesized on said bead; and

b) synthesizing said entities on said beads, wherein said beads are sorted after each synthesis cycle according to said optical labeling substances,

whereby a library is synthesized, wherein each of said beads contains an entity that corresponds to an optical labeling substance on said bead and the sum of said beads collectively contains a plurality of entities that is predetermined before the library synthesis.

69. The method of claim 68, wherein the optical labeling substance is a quantum dot.

70. The method of claim 68, wherein the beads further comprises an element that facilitates and/or enables manipulation of the beads and/or the bead/synthesized entity complexes.

71. The method of claim 70, wherein the element is selected from the group consisting of a cell, a cellular organelle, a virus, a microparticle, an aggregate or complex of molecules and an aggregate or complex thereof.

72. The method of claim 68, wherein the beads further comprise a molecular tag.

73. The method of claim 72, wherein the molecular tag is a DNA sequence or an antibody.

74. The method of claim 68, wherein each of the beads contains a single synthesized entity.

75. The method of claim 68, wherein the synthesized entities are selected from group consisting of peptides, proteins, oligonucleotides, nucleic acids, vitamins, oligosaccharides, carbohydrates, lipids, small molecules, or a complex or combination thereof.

76. The method of claim 68, wherein the synthesized library comprises a defined set of entities that are involved in a biological pathway, belongs to a group of entities with identical or similar biological function, expressed in a stage of cell cycle, expressed in a cell type, expressed in a tissue type, expressed in an organ type, expressed in a developmental stage, entities whose expression and/or activity are altered in a disease or disorder type or stage, or entities whose expression and/or activity are altered by drug or other treatments.

77. The method of claim 68, wherein the synthesized library comprises a defined set of DNA fragments.

78. The method of claim 77, wherein each of the DNA fragments in the synthesized library comprises at least 10, 15, 20, 25, 50, 75, 100, 200 or 500 nucleotides.

79. The method of claim 68, wherein the synthesized library comprises a defined set of protein or peptide fragments.

80. The method of claim 79, wherein each of the protein or peptide fragments in the synthesized library comprises at least 10, 15, 20, 25, 50, 75, 100, 200 or 500 amino acids.

81. A library that is synthesized according to the method of claim 68.

82. A method for generating an antibody library, which method comprises:

a) contacting the library of claim 81 with a plurality of antibodies;

b) selecting and/or recovering the antibodies that specifically bind to the entities of the library of claim 81.

83. The method of claim 82, wherein the plurality of antibodies is derived from a phage display library.

86. A kit for manipulating a moiety, which kit comprises:

a) a bead of claim 1 comprising a binding partner that is capable of binding to a moiety to be manipulated; and

b) a means for generating a physical force for manipulating said moiety-bead complex.

87. The kit of claim 86, wherein the bead comprises an optical labeling substance.

88. The kit of claim 87, wherein the optical labeling substance is a quantum dot.

84. A method for synthesizing a library, which method comprises:

a) providing a plurality of beads, each of said beads comprising a magnetizable substance, an electrically conductive substance and a unique optical labeling substance; and

b) synthesizing an entity on said beads, wherein said beads are identified after each synthesis cycle according to said unique optical labeling substances,

whereby a library is synthesized, wherein each of said beads contains an entity that can be identified according to said unique optical labeling substance on each of said beads.

85. A library that is synthesized according to the method of claim 84.