초록

The invention provides an electrophoresis cassette, methods for making the electrophoresis cassette, and method of fractionating analytes from a sample based upon electrophoretic mobility in a single application of the sample to an electrophoretic system.

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1. An electrophoresis cassette comprising: a plate including at least one macrofluidic separation channel, the channel having a first physically and electrically isolated portion and a second physically and electrically isolated portion;an elution chamber positioned on one or another of the physical

1. An electrophoresis cassette comprising: a plate including at least one macrofluidic separation channel, the channel having a first physically and electrically isolated portion and a second physically and electrically isolated portion;an elution chamber positioned on one or another of the physically and electrically isolated portions, the chamber comprising at least one an elution cavity and an analyte-impermeable barrier. 2. The cassette of claim 1, wherein the elution chamber further comprises at least one of an analyte-permeable barrier, a sample collection chamber including a sample removal port, and an analyte-impermeable barrier. 3. The cassette of claim 1, further comprising a buffer reservoir for each of the macrofluidic separation channels. 4. The cassette of claim 1, further comprising a cover for the plate. 5. The cassette of claim 3, further comprising at least one of a sample well cavity, an elution reservoir, and a waste reservoir. 6. The cassette of claim 5, further comprising a division point provided between the macrofluidic channel and the elution reservoir and the waste reservoir. 7. The cassette of claim 5, wherein the elution reservoir and the waste reservoir are provided at an end of the first physically and electrically isolated portions and the second physically and electrically isolated portions, respectively. 8. The cassette of claim 1, further comprising a cover for the plate, wherein the cover includes a configuration that corresponds to the configuration of the top of the plate. 9. The cassette of claim 8, wherein the cover includes at least one of an opening, a protrusion and a recess that align with at least one of the macrofluidic channel, the sample well cavity, and the sample removal port. 10. The cassette of claim 8, wherein the cover includes at least one of an electrode port, a vent, and an injection port. 11. The cassette of claim 1, wherein the separation channel is tapered from one end to the constriction point. 12. The cassette of claim 6, wherein the constriction point is the division point. 13. The cassette of claim 1, wherein the cassette is disposable. 14. The cassette of claim 5, wherein the cassette further comprises: a negative electrode positioned between the buffer reservoir and a corresponding end of the separation channel;a positive electrode positioned between an end of the first physically and electrically isolated portion and the elution reservoir; anda positive electrode positioned between an end of the second physically and electrically isolated portion and the waste reservoir. 15. The cassette of claim 5, wherein the cavity for the first dam is positioned between the buffer reservoir and the sample well cavity. 16. The cassette of claim 6, wherein the cavity for the second dam is positioned between the division point and the waste reservoir. 17. The cassette of claim 16, wherein the at least one of a first dam or a second dam is inserted into either the cavity for the first dam or the cavity for the second dam, respectively. 18. The cassette of claim 17, wherein the at least one of the first dam or the second dam comprises a frame attached to an ion-permeable barrier. 19. The cassette of claim 18, wherein the ion-permeable barrier comprises a least one pore having a diameter range of between 0.001 micron to 0.1 micron. 20. The cassette of claim 1, wherein the microfluidic channel includes at least one of a gel matrix composition, a liquid buffer composition, and a solid buffer composition. 21. The cassette of claim 1, wherein at least one of a gel matrix composition, a liquid buffer composition, and a solid buffer composition comprises at least one of a fluorophore or a chromophore. 22. The cassette of claim 6, wherein at least one macrofluidic separation channel further comprises a gel matrix composition. 23. The cassette of claim 22, wherein the gel matrix composition fills a volume of the macrofluidic separation channel, including at least one of the first and second physically and electrically isolated portions. 24. The cassette of claim 22, wherein the gel matrix composition defines at least one sample well within at least one sample well cavity. 25. The cassette of claim 24, wherein the sample well forms a gel chimney. 26. The cassette of claim 22, wherein at least one macrofluidic channel further comprises a buffer composition. 27. The cassette of claim 26, wherein the buffer composition fills a volume of at least one buffer reservoir, at least one sample well, at least one elution reservoir, and at least one waste reservoir. 28. The cassette of claim 26, wherein at least one elution chamber further comprises an elution buffer composition. 29. The cassette of claim 28, wherein the elution buffer composition fills a volume of at least one elution chamber. 30. The cassette of claim 28, wherein the cassette further comprises a removable seal. 31. The cassette of claim 30, wherein the seal encloses at least one of an opening, a protrusion and a recess of a cover. 32. The cassette of claim 1, wherein at least one macrofluidic separation channel is optically-transparent. 33. The cassette of claim 32, wherein the separation channel is optically-transparent on at least one side. 34. The cassette of claim 32, wherein the separation channel is optically-transparent on only one side. 35. The cassette of claim 1, wherein the cassette comprises between 1 and 5 macrofluidic separation channels. 36. The cassette of claim 1, wherein the cassette comprises between 1 and 9 macrofluidic separation channels. 37. The cassette of claim 1, wherein the cassette comprises between 1 and 13 macrofluidic separation channels. 38. The cassette of claim 6, wherein the macrofluidic channel further comprises a lens positioned between the sample well cavity and the division point. 39. The cassette of claim 38, wherein said the lens comprises a gel matrix composition. 40. The cassette of claim 1, wherein the analyte-permeable barrier of the elution chamber comprises a hydrophilic membrane or filter. 41. The cassette of claim 40, wherein the analyte-permeable barrier comprises a least one pore having a diameter range of between 0.4 micron to 50 microns. 42. The cassette of claim 40, wherein the analyte-permeable barrier comprises a least one pore having a diameter range of between 0.4 micron to 1 micron. 43. The cassette of claim 1, wherein the analyte-impermeable barrier of the elution chamber is a membrane, filter, film, or any combination thereof. 44. The cassette of claim 43, wherein the analyte-impermeable barrier is an ultrafiltration membrane or a conductive film. 45. The cassette of claim 44, wherein the ultrafiltration membrane comprises a least one pore having a diameter range of between 0.001 micron to 0.1 micron. 46. The cassette of claim 44, wherein the ultrafiltration membrane has a molecular weight cutoff of between 1,000 to 30,000 daltons. 47. The cassette of claim 44, wherein the ultrafiltration membrane has a molecular weight cutoff of between 3,000 to 10,000 daltons. 48. The cassette of claim 44, wherein the analyte-impermeable barrier comprises a conductive film having the same charge as the analyte. 49. The cassette of claim 44, wherein the analyte-impermeable barrier comprises a conductive film contacted with negatively-charged sulfate groups. 50. The cassette of claim 44, wherein the analyte-impermeable barrier is Nafion. 51. A method of making an electrophoresis cassette, comprising: providing an electrophoresis cassette of claim 1, wherein the cassette further comprises a sample well insert and a cover, wherein the sample well insert traverses an opening in the cover plate aligned with the sample well cavity;inserting a gel matrix composition through an injection port;solidifying the gel matrix composition, wherein the gel matrix composition transforms from a liquid to a solid;removing the sample well insert, wherein a sample well is generated;filling the buffer reservoir, the elution reservoir, and the waste reservoir with a buffer composition;filling the elution chamber with an elution buffer composition; andsealing the electrophoresis cassette. 52. The method of claim 51, wherein the electrophoresis cassette is horizontal during the inserting and solidifying steps. 53. The method of claim 51, wherein the electrophoresis cassette comprises between 1 and 5 macrofluidic separation channels. 54. The method of claim 51, wherein the electrophoresis cassette comprises between 1 and 9 macrofluidic separation channels. 55. The method of claim 51, wherein the electrophoresis cassette comprises between 1 and 13 macrofluidic separation channels. 56. A detection system for detecting a property of an analyte within a sample comprising: a cassette according to claim 1; an electrode array comprising at least one of a negative electrode and a positive electrode, wherein the negative electrode aligns with a position on the cassette between the buffer reservoir and a corresponding end of the separation channel, and wherein the positive electrode aligns with a physically and electrically isolated portion of the separation channel;a detector positioned near the separation channel of the electrophoresis cassette, wherein the detector detects a property of an analyte;a processor configured to activate or deactivate power to at least one positive electrode based upon a signal received from the detector; anda power module comprising at least one of a power supply and a relay to provide power to at least one of the processor, the negative electrode and at least one positive electrode. 57. The system of claim 56, wherein the property is an optical property of an analyte. 58. The system of claim 56, wherein the sample comprises a fluorescent compound and the analyte forms a complex with the fluorescent compound. 59. The system of claim 56, wherein the fluorescent compound is a fluorophore. 60. The system of claim 56, wherein the sample comprises a light-absorbing compound and the analyte forms a complex with the light-absorbing compound. 61. The system of claim 60, wherein the light-absorbing compound is a chromophore. 62. The system of claim 57, wherein the optical property is the emission of light. 63. The system of claim 57, wherein the optical property is the absorption of light. 64. The system of claim 56, wherein the analyte is a sample or a molecular weight marker. 65. The system of claim 64, wherein the detector detects a property of the molecular weight marker within a first microfluidic channel and send a signal to the processor. 66. The system of claim 65, wherein the processor receives the signal from the detector and applies an algorithm to determine the molecular weight of at least one of an analyte at the division point of a second macrofluidic channel. 67. A method of fractionating analytes within a sample, comprising: providing an electrophoresis cassette of claim 1, wherein the cassette further comprises a sample well insert and a cover, wherein the sample well insert traverses an opening in the cover plate aligned with the sample well cavity;inserting a gel matrix composition through an injection port;solidifying the gel matrix composition, wherein the gel matrix composition transforms from a liquid to a solid;removing the sample well insert, wherein a sample well is generated;filling the buffer reservoir, the elution reservoir, and the waste reservoir with a buffer composition;filling the elution chamber with an elution buffer composition; and inserting the electrophoresis cassette into a detection system;programming the processor of the detection system to selectively activate the positive electrode of the electrode array aligned with the physically and electrically isolated portion of the separation channel comprising the elution chamber when the processor determines that at least one of an analyte of the desired molecular weight is traversing the division point of the separation channel;applying the sample to the sample well;applying a voltage across the electrophoresis cassette;collecting analytes of the sample having a desired electrophoretic mobility in the elution chamber,thereby fractionating analytes within a sample. 68. The method of claim 67, wherein the sample comprises a detectable compound. 69. The method of claim 67, wherein the sample comprises at least one of a complex of an analyte and a fluorescent compound. 70. The method of claim 69, wherein the fluorescent compound is a fluorophore. 71. The method of claim 67, wherein the sample comprises at least one of a complex of an analyte and a light-absorbing compound. 72. The method of claim 71, wherein the light-absorbing compound is a chromophore. 73. The method of claim 67, wherein at least one of the gel matrix composition, the buffer composition, or the elution buffer composition comprises at least one of a fluorophore that complexes to at least one of an analyte. 74. The method of claim 67, wherein at least one of the gel matrix composition, the buffer composition, or the elution buffer composition comprises at least one of a chromophore that complexes to at least one of an analyte. 75. The method of claim 67, wherein the sample comprises a molecular weight marker. 76. The method of claim 67, wherein the processor of the detection system selectively activates the positive electrode of the electrode array aligned with the physically and electrically isolated portion of the separation channel comprising the elution chamber when an analyte having a specified electrophoretic mobility is detected and wherein the specified electrophoretic mobility is distinct for each macrofluidic channel. 77. The method of claim 67, wherein the analyte is a polynucleic acid or a polypeptide. 78. The method of claim 77, wherein the polynucleic acid comprises deoxyribonucleic acid (DNA) or ribonucleic acid (RNA). 79. The method of claim 77, wherein the polynucleic acid is double or single stranded. 80. The method of claim 77, wherein the polypeptide is native or denatured. 81. A device comprising an electrophoresis cassette, the electrophoresis cassette including: (a) a channel plate comprising a macrofluidic channel, wherein the macrofluidic channel comprises, from proximal to distal, a buffer reservoir, a first end of a separation channel, a cavity for a first dam, a sample well cavity, a constriction point, a division point, an elution chamber cavity, a second physically and electrically isolated end of the separation channel, a third physically and electrically isolated end of the separation channel, an elution reservoir, a cavity for a second dam, and a waste reservoir;(b) an elution chamber comprising, from proximal to distal, an analyte-permeable barrier, a sample collection chamber having a sample removal port, and an analyte-impermeable barrier, wherein the elution module is attached to the elution chamber cavity;(c) a cover plate that contacts the top of the channel plate, wherein the cover plate comprises at least one of an opening, a protrusion, and a recess that align, from proximal to distal, with the sample well cavity and the elution chamber;(d) a gel matrix composition that fills the macrofluidic separation channel and defines a sample well within the sample well cavity;(e) a liquid buffer composition that fills the buffer reservoir, the sample well, the elution reservoir, and the waste reservoir;(f) an elution buffer composition that fills the elution chamber; and(g) a seal that encloses the electrophoresis cassette. 82. The device of claim 81, wherein the cover comprises at least one of an electrode port, a vent, and an injection port. 83. The device of claim 81, wherein at least one of the cavity for a first dam or the cavity for a second dam comprises a first dam or a second dam, respectively. 84. The device of claim 81, wherein the sample well forms a gel chimney.