High throughput screening of crystallization of materials

• US 20050178317A1
• Filed: 04/12/2005
• Published: 08/18/2005
• Est. Priority Date: 04/05/2001
• Status: Active Grant

• Current Assignee: California Institute of Technology
• Original Assignee: California Institute of Technology
• Inventors: Hansen, Carl I., Quake, Stephen R., Berger, James M.
• Granted Patent: US 7,670,429 B2
• Time in Patent Office: Days
• US Class Current: 117/68
• CPC Class Codes: B01J 2219/00355 peristaltic
• CPC Class Codes: B01J 2219/00378 Piezoelectric or ink jet dispensers
• CPC Class Codes: B01J 2219/00396 Membrane valves
• CPC Class Codes: B01J 2219/00398 in multiple arrangements
• CPC Class Codes: B01J 2219/00439 using micromirror arrays
• CPC Class Codes: B01J 2219/005 Beads
• CPC Class Codes: B01J 2219/00527 Sheets
• CPC Class Codes: B01J 2219/00605 the compounds being directly bound or immobilised to solid supports
• CPC Class Codes: B01J 2219/0061 The surface being organic
• CPC Class Codes: B01J 2219/00621 by physical means, e.g. trenches, raised areas
• CPC Class Codes: B01J 2219/00637 by coating it with another layer
• CPC Class Codes: B01J 2219/00659 Two-dimensional arrays
• CPC Class Codes: B01J 2219/00707 separated from the reactor apparatus
• CPC Class Codes: B01J 2219/00722 Nucleotides
• CPC Class Codes: B01J 2219/00725 Peptides
• CPC Class Codes: B01J 2219/00756 Compositions, e.g. coatings, crystals, formulations
• CPC Class Codes: B01L 2200/025 Align devices or objects to ensure defined positions relative to each other
• CPC Class Codes: B01L 2200/027 for microfluidic devices
• CPC Class Codes: B01L 2200/0605 Metering of fluids
• CPC Class Codes: B01L 2200/10 Integrating sample preparation and analysis in single entity, e.g. lab-on-a-chip concept
• CPC Class Codes: B01L 2300/0681 : Filter
• CPC Class Codes: B01L 2300/0861 : Configuration of multiple channels and/or chambers in a single devices
• CPC Class Codes: B01L 2300/123 : Flexible; Elastomeric
• CPC Class Codes: B01L 2300/14 : Means for pressure control
• CPC Class Codes: B01L 2300/18 : Means for temperature control
• CPC Class Codes: B01L 2400/0481 : squeezing of channels or chambers
• CPC Class Codes: B01L 2400/0655 : pinch valves
• CPC Class Codes: B01L 2400/0688 : surface tension valves, capillary stop, capillary break
• CPC Class Codes: B01L 3/06 : Crystallising dishes
• CPC Class Codes: B01L 3/50273 : characterised by the means or forces applied to move the fluids micropumps F04B19/006, of the membrane type F04B43/043
• CPC Class Codes: B01L 3/502738 : characterised by integrated valves microvalves F16K99/0001
• CPC Class Codes: B01L 7/54 : using spatial temperature gradients
• CPC Class Codes: B01L 9/527 : for microfluidic devices, e.g. used for lab-on-a-chip
• CPC Class Codes: C30B 29/54 : Organic compounds

Abstract

High throughput screening of crystallization of a target material is accomplished by simultaneously introducing a solution of the target material into a plurality of chambers of a microfabricated fluidic device. The microfabricated fluidic device is then manipulated to vary the solution condition in the chambers, thereby simultaneously providing a large number of crystallization environments. Control over changed solution conditions may result from a variety of techniques, including but not limited to metering volumes of crystallizing agent into the chamber by volume exclusion, by entrapment of volumes of crystallizing agent determined by the dimensions of the microfabricated structure, or by cross-channel injection of sample and crystallizing agent into an array of junctions defined by intersecting orthogonal flow channels.

Claims

1-32. -32. (canceled)

33. A method for determining crystallization conditions for a protein, the method comprising:

within a microfluidic device, delivering material to an enclosed microvolume via one or more lumens that each have a cross sectional diameter of less than 500 microns to form a plurality of different crystallization samples within the enclosed microvolume, the plurality of different crystallization samples comprising a protein to be crystallized and crystallization conditions which vary among the plurality of different crystallization samples;

allowing crystals of the protein to form in the plurality of crystallization samples within the microfluidic device; and

identifying which of the plurality of crystallization samples within the microfluidic device comprise a precipitate or a crystal of the protein.

34. A method according to claim 33 wherein the enclosed microvolume is a lumen.

35. A method according to claim 33 wherein the enclosed microvolume is a lumen with a cross sectional diameter of less than 2.5 mm.

36. A method according to claim 33 wherein the enclosed microvolume is a lumen with a cross sectional diameter of less than 1 mm.

37. A method according to claim 33 wherein the enclosed microvolume is a lumen with a cross sectional diameter of less than 500 microns.

38. A method according to claim 33 wherein the enclosed microvolume is a microchamber.

39. A method according to claim 33 wherein the enclosed microvolume is at least partially enclosed within a substrate which comprises other enclosed microvolumes which also comprise crystallization samples.

40. A method according to claim 33 wherein the enclosed microvolume is at least partially enclosed within a card shaped substrate.

41. A method according to claim 33, the method further comprising performing a spectroscopic analysis on a precipitate or crystal formed within the microvolume.

42. A method according to claim 41, wherein the spectroscopic analysis is selected from the group consisting of Raman, UV/VIS, IR, and x-ray spectroscopy.

43. A method according to claim 41, wherein the spectroscopic analysis is x-ray spectroscopy.

44. A method according to claim 43, wherein x-ray spectroscopy is performed such that a portion of the microvolume that the x-ray beam traverses contains at least as many electrons as is contained in a material defining the portion of the microvolume that the x-ray beam traverses.

45. A method according to claim 43, wherein x-ray spectroscopy is performed such that a portion of the microvolume that the x-ray beam traverses contains at least three times as many electrons as is contained in a material defining the portion of the microvolume that the x-ray beam traverses.

46. A method according to claim 43, wherein x-ray spectroscopy is performed such that a portion of the microvolume that the x-ray beam traverses contains at least five times as many electrons as is contained in a material defining the portion of the microvolume that the x-ray beam traverses.

47. A method according to claim 43, wherein x-ray spectroscopy is performed such that a portion of the microvolume that the x-ray beam traverses contains at least ten times as many electrons as is contained in a material defining the portion of the microvolume that the x-ray beam traverses.

48. A method according to claim 33, wherein material defining the microvolume defines a groove that reduces a number of electrons that an x-ray beam used to perform x-ray spectroscopy of a crystal within the microvolume traverses in the process of performing x-ray spectroscopy on the sample within the microvolume.

49. A method according to claim 33, wherein the method further comprises forming the plurality of different crystallization samples within the enclosed microvolume.

50. A method according to claim 33, wherein one or more dividers are positioned within the enclosed microvolume to separate adjacent crystallization samples within the enclosed microvolume.

51. A method according to claim 50, wherein the one or more dividers are formed of an impermeable material.

52. A method according to claim 50, wherein the impermeable material is an impermeable liquid.

53. A method according to claim 50, wherein the impermeable material is an impermeable solid.

54. A method according to claim 50, wherein the one or more dividers are formed of a permeable material.

55. A method according to claim 50, wherein the one or more dividers are formed of a semipermeable material.

59. A method according to claim 50, wherein at least one of the one or more dividers form an interface selected from the group consisting of liquid/liquid, liquid/gas interface, liquid/solid and liquid/sol-gel interface.

60. A method according to claim 50, wherein the one or more dividers are selected from the group consisting of a membrane, gel, frit, and matrix.

61. A method according to claim 50, wherein the one or more dividers function to modulate diffusion characteristics between adjacent crystallization samples.

62. A method according to claim 50, wherein at least one of the one or more dividers is formed of a semipermeable material which allows diffusion between adjacent crystallization samples.

64. A method according to claim 43, wherein the x-ray spectroscopy is x-ray diffraction.

65. A method according to claim 43, wherein x-ray spectroscopy is performed such that a portion of the crystal or precipitate that the x-ray beam traverses contains at least as many electrons as is otherwise traversed by the x-ray beam when traversing a device comprising the microvolume.

66. A method according to claim 43, wherein x-ray spectroscopy is performed such that a portion of the crystal or precipitate that the x-ray beam traverses contains at least three times as many electrons as is otherwise traversed by the x-ray beam when traversing a device comprising the microvolume.

67. A method according to claim 43, wherein x-ray spectroscopy is performed such that a portion of the crystal or precipitate that the x-ray beam traverses contains at least five times as many electrons as is otherwise traversed by the x-ray beam when traversing a device comprising the microvolume.

68. A method according to claim 43, wherein x-ray spectroscopy is performed such that a portion of the crystal or precipitate that the x-ray beam traverses contains at least ten times as many electrons as is otherwise traversed by the x-ray beam when traversing a device comprising the microvolume.

70. A method according to claim 43, wherein x-ray spectroscopy is performed such that a portion of the microvolume that the x-ray beam traverses contains at least half as many electrons as is contained in a material defining the portion of the microvolume that the x-ray beam traverses.

56. A method according to claim 74, wherein the semipermeable material is a gas.

57. A method according to claim 74, wherein the semipermeable material is a liquid.

58. A method according to claim 74, wherein the semipermeable material is a gel.

63. A method for determining crystallization conditions for a protein, the method comprising:

within a microfluidic device, delivering material to a plurality of enclosed microvolumes via one or more lumens that each have a cross sectional diameter of less than 500 microns to form a plurality of different crystallization samples within the plurality of enclosed microvolumes, each microvolume comprising two or more crystallization samples, the different crystallization samples comprising a protein to be crystallized and crystallization conditions which vary among the plurality of different crystallization samples;

allowing crystals of the protein to form in the plurality of crystallization samples; and

identifying which of the plurality of crystallization samples comprise a precipitate or a crystal of the protein.

69. A method according to claim 63, wherein each microvolume comprising a plurality of crystallization samples.

80. A method for determining crystallization conditions for a protein, the method comprising:

delivering material within a microfluidic device to an enclosed volume via one or more flow channels that each have a width and depth of less than 500 microns to form a plurality of different crystallization samples within the enclosed volume,

the plurality of different crystallization samples comprising a protein to be crystallized and crystallization conditions which vary among the plurality of different crystallization samples;

allowing crystals of the protein to form in the plurality of crystallization samples within the microfluidic device; and

identifying which of the plurality of crystallization samples within the microfluidic device comprise a precipitate or a crystal of the protein.

81. A method for determining crystallization conditions for a protein, the method comprising:

within a microfluidic device, delivering material to a plurality of enclosed volumes via one or more flow channels that each have a cross sectional diameter of less than 500 microns to form a plurality of different crystallization samples within the plurality of enclosed volumes, each enclosed volume comprising two or more crystallization samples, the different crystallization samples comprising a protein to be crystallized and crystallization conditions which vary among the plurality of different crystallization samples;

allowing crystals of the protein to form in the plurality of crystallization samples; and

identifying which of the plurality of crystallization samples comprise a precipitate or a crystal of the protein.