Electrophoresis apparatus having staggered passage configuration

• US 7,329,388 B2
• Filed: 12/05/2003
• Issued: 02/12/2008
• Est. Priority Date: 11/08/1999
• Status: Expired due to Term

• Current Assignee: Princeton Biochemicals, Inc.
• Original Assignee: Princeton Biochemicals, Inc.
• Inventors: Guzman, Norberto A.
• Primary Examiner(s): Le; Long V.
• Assistant Examiner(s): Jung; Unsu
• Application Number:
  US10/728,499
• Publication Number:
  US 20050155861A1
• Time in Patent Office: 1,530 Days
• Field of Search: 435/4, 435/6, 435/7.2, 435/7.21, 435/287.1, 435/287.2, 435/287.9, 4352884-2885, 435/288.7, 435/286.5, 435/306.1, 435/372, 436/164, 436/514, 436/518, 436/524, 436/527, 436/531, 436/533, 436/534, 422 55- 58, 204/180.1, 204450-455, 204600-605
• US Class Current: 422/82.01
• CPC Class Codes: G01N 27/44726 using specific dyes, markers or binding molecules
• CPC Class Codes: G01N 27/44743 Introducing samples
• CPC Class Codes: G01N 27/44773 Multi-stage electrophoresis, e.g. two-dimensional electrophoresis
• CPC Class Codes: Y10T 436/25375 Liberation or purification of sample or separation of material from a sample [e.g., filtering, centrifuging, etc.]
• CPC Class Codes: Y10T 436/255 including use of a solid sorbent, semipermeable membrane, or liquid extraction

Abstract

An electrophoresis apparatus is generally disclosed for sequentially analyzing a single sample or multiple samples having one or more analytes in high or low concentrations. The apparatus comprises a relatively large-bore transport capillary which intersects with a plurality of small-bore separation capillaries and includes a valve system. Analyte concentrators, having antibody-specific (or related affinity) chemistries, are stationed at the respective intersections of the transport capillary and separation capillaries to bind one or more analytes of interest. The apparatus allows the performance of two or more dimensions for the optimal separation of analytes. The apparatus may also include a plurality of valves surrounding each of the analyte concentrators to localize each of the concentrators to improve the binding of one or more analytes of interest.

Claims

1. An electrophoresis apparatus, comprising:

a transport capillary or channel;

a first separation capillary or channel overlapping and intersecting the transport capillary or channel at a first intersection;

the first intersection having a first staggered configuration which includes a first elongated concentration area;

a first analyte concentrator containing at least one first immobilized affinity ligand in the first elongated concentration area to concentrate a first analyte of interest from a sample introduced into the transport capillary or channel;

a second separation capillary or channel overlapping and intersecting the transport capillary or channel at a second intersection spaced downstream from the first intersection;

the second intersection having a second staggered configuration which includes a second elongated concentration area;

a second analyte concentrator containing at least one second immobilized affinity ligand in the second elongated concentration area to concentrate a second analyte of interest from the sample introduced into the transport capillary or channel;

analyte detector means for identifying and characterizing the first and second analytes of interest conveyed thereto from the first and second analyte concentrators, respectively; and

controlling means for controlling flow of the sample in the transport capillary or channel and past the first and second intersections and for controlling flow of buffer fluid through the first and second separation capillaries or channels and conveyed by electrophoresis migration, pressure or a combination of electrophoresis migration and pressure to the analyte detector means.

2. The electrophoresis apparatus of claim 1 wherein the first analyte concentrator includes a matrix assembly having a surface to which the first immobilized affinity ligand is bound.

3. The electrophoresis apparatus of claim 2 wherein the matrix assembly includes a plurality of microstructures.

4. The electrophoresis apparatus of claim 2 wherein the matrix assembly is free-floating, and the first analyte concentrator retains the free-floating matrix assembly by pressure-resistant porous end walls or frits disposed in the transport capillary or channel and the first separation capillary or channel.

5. The electrophoresis apparatus of claim 2 wherein the matrix assembly includes a fixed architecture defined by beaded microstructures interconnected to each other and to an inner wall of the first elongated concentration area.

6. The electrophoresis apparatus of claim 2 wherein the matrix assembly includes a fixed architecture fabricated from polymeric microstructures interconnected to each other and to the first elongated concentration area.

7. The electrophoresis apparatus of claim 1 further comprising an auxiliary capillary through which a cleaning solution and a separation buffer can be introduced into the first separation capillary or channel downstream of the first analyte concentrator.

8. The electrophoresis apparatus of claim 7 wherein the auxiliary capillary is an electrolyte-provider and cleaning solution-provider capillary.

9. The electrophoresis apparatus of claim 1 further comprising an auxiliary capillary coupled to the second separation capillary or channel downstream of the second analyte concentrator to provide a buffer fluid, including a cleaning solution and a separation buffer, to the second separation capillary or channel away from the second analyte concentrator.

10. The electrophoresis apparatus of claim 9 wherein the controlling means controls flow in the auxiliary capillary.

11. The electrophoresis apparatus of claim 1 further comprising an auxiliary analyte concentrator on the first separation capillary or channel and downstream of the first analyte concentrator, the auxiliary analyte concentrator having at least one affinity ligand capable of retaining chromophores to bind the first analyte of interest released from the first analyte concentrator to improve the sensitivity and selectivity of the first analyte of interest.

12. The electrophoresis apparatus of claim 1 wherein the first separation capillary or channel is filled with an electrically conductive fluid.

13. The electrophoresis apparatus of claim 1 wherein the first separation capillary or channel is filled with a gel matrix and an electrically conductive fluid.

14. The electrophoresis apparatus of claim 1 wherein the sample has a plurality of proteins with different isoelectric point levels, which are further separated through the separation capillaries or channels by at least one mode of capillary electrophoresis after being subjected to isoelectric focusing in the transport capillary or channel.

15. The electrophoresis apparatus of claim 1 wherein at least one of the first and/or second immobilized affinity ligands is capable of performing at least one chemical or biochemical reaction.

16. The electrophoresis apparatus of claim 15 wherein the at least one chemical or biochemical reaction includes peptide synthesis, nucleic acid synthesis, or an enzymatic reaction.

17. The electrophoresis apparatus of claim 1 wherein at least one of the analyte concentrators has an encapsulated cellular or subcellular structure.

18. The electrophoresis apparatus of claim 17 wherein the encapsulated cellular or subcellular structure is adapted for drug metabolism studies and/or for metabolic pathway studies.

19. The electrophoresis apparatus of claim 1 wherein at least one of the analyte concentrators has an acoustic micromixing system.

20. The electrophoresis apparatus of claim 1 wherein at least one of the analyte concentrators has a microwave pulse system.

21. The electrophoresis apparatus of claim 1 wherein the first immobilized affinity ligand is covalently bound to a matrix assembly of the first analyte concentrator.

22. The electrophoresis apparatus of claim 1 further comprising separation buffer fluid means for providing a separation buffer fluid to the first separation capillary or channel and downstream of the first analyte concentrator and to the second separation capillary or channel and downstream of the second analyte concentrator.

23. The electrophoresis apparatus of claim 22 wherein the controlling means controls the operation of the separation buffer fluid means.

24. The electrophoresis apparatus of claim 22 wherein the separation buffer fluid contains sodium tetraborate.

25. The electrophoresis apparatus of claim 22 wherein the separation buffer fluid includes an organic solvent, or a mixture of organic solvents and additives.

26. The electrophoresis apparatus of claim 1 wherein the controlling means controls the flow of cleaning buffers and separation buffers in the separation capillaries or channels.

27. The electrophoresis apparatus of claim 1 further comprising a first electrolyte-provider capillary or channel in fluid communication with the first separation capillary or channel downstream of the first analyte concentrator and a second electrolyte-provider capillary or channel in fluid communication with the second separation capillary or channel downstream of the second analyte concentrator.

28. The electrophoresis apparatus of claim 1 wherein the analyte detector means is an ultraviolet detector system.

29. The electrophoresis apparatus of claim 1 wherein the analyte detector means is a fluorescence or laser-induced fluorescence detector system.

30. The electrophoresis apparatus of claim 1 wherein the analyte detector means is a conductivity, electrochemical, radioactive, mass spectrometer, circular diochroism or nuclear magnetic resonance detector system.

31. The electrophoresis apparatus of claim 1 wherein the analyte detector means includes a combination of several detectors used simultaneously.

32. The electrophoresis apparatus of claim 1 wherein the first and second separation capillaries or channels merge into a single exit output capillary or channel.

33. The electrophoresis apparatus of claim 32 wherein the controlling means controls, at the merging of the first and second separation capillaries or channels, sequential fluid flow from the first and second separation capillaries or channels to the exit output capillary or channel.

34. The electrophoresis apparatus of claim 1 further comprising an exit outlet capillary or channel into which the first and second separation capillaries or channels flow and at a detection zone of the analyte detector means.

35. The electrophoresis apparatus of claim 34 wherein the controlling means controls the sequential fluid flow of the first and second separation capillaries or channels to the detection zone.

36. The electrophoresis apparatus of claim 34 further comprising a grounding electrode at an outlet end of the exit outlet capillary or channel.

37. The electrophoresis apparatus of claim 36 wherein the exit outlet capillary or channel flows into a container in which the grounding electrode is positioned.

38. The electrophoresis apparatus of claim 1 wherein the first and second separation capillaries or channels have separate output capillaries or channels, and the analyte detector means includes a first analyte detector for the first separation capillary or channel and a separate second analyte detector for the second separation capillary or channel.

39. The electrophoresis apparatus of claim 1 wherein the first and second separation capillaries or channels have separate output capillaries or channels and the analyte detector means includes a detector which is movable between the first and second separation capillaries or channels.

40. The electrophoresis apparatus of claim 1 further comprising capillary or channel bulging members which retain the at least one first immobilized affinity ligand to a support matrix in the first analyte concentrator.

41. The electrophoresis apparatus of claim 1 wherein the inner diameter of the transport capillary or channel is larger than the inner diameter of the first separation capillary or channel and than the inner diameter of the second separation capillary or channel.

42. The electrophoresis apparatus of claim 1 wherein the transport capillary or channel and the first and second separation capillaries or channels are all capillaries.

43. The electrophoresis apparatus of claim 1 wherein the transport capillary or channel and the first and second separation capillaries or channels are all channels.

44. The electrophoresis apparatus of claim 1 wherein the electrophoresis apparatus is a capillary electrophoresis apparatus.

45. The electrophoresis apparatus of claim 1 wherein the electrophoresis apparatus is a microchip electrophoresis apparatus.

46. The electrophoresis apparatus of claim 1 wherein the first immobilized affinity ligands are covalently linked to an inner wall of the first staggered configuration.

47. The electrophoresis apparatus of claim 1 wherein the controlling means includes transport capillary or channel valves and separation capillary or channel valves, and wherein the transport capillary or channel valves are adapted to be opened and the first separation capillary or channel valves are adapted to be closed to allow fluid to pass through the first analyte concentrator towards an outlet end of the transport capillary or channel.

48. The electrophoresis apparatus of claim 47 wherein the fluid is the sample, at least one chromophoric substance, and at least one cleaning buffer.

49. The electrophoresis apparatus of claim 1 wherein the controlling means includes transport capillary or channel valves and first separation capillary or channel valves, and wherein the transport capillary or channel valves are adapted to be closed and the first separation capillary or channel valves are adapted to be opened to allow a separation buffer solution to pass through the first analyte concentrator and in the first separation capillary or channel to the analyte detector means.

50. The electrophoresis apparatus of claim 1 wherein the positive electrode of the electrophoresis apparatus is generally at an inlet end of the separation capillaries or channels and the negative electrode is downstream of the analyte concentrators.

51. The electrophoresis apparatus of claim 1 wherein an inlet end of the first separation capillary or channel is alternatively in fluid communication with a cleaning solution supply, or an eluting buffer supply or a separation buffer supply.

52. The electrophoresis apparatus of claim 1 wherein the capillaries or channels are fused-silica or plastic tubes or glass or plastic channels.

53. The electrophoresis apparatus of claim 1 further comprising a buffer supply for the first separation capillary or channel and which includes a separation buffer and an eluting buffer to release the bound first analyte of interest from the at least one first immobilized affinity ligand.

54. The electrophoresis apparatus of claim 1 wherein the controlling means include valves on the transport capillary or channel on opposite sides of the first analyte concentrator and valves on the first separation capillary or channel on opposite sides of the first analyte concentrator.

55. The electrophoresis apparatus of claim 1 wherein the first separation capillary or channel is positioned and capable of separating therein the first analyte of interest retained by the first immobilized affinity ligand after the first analyte is released from the first immobilized affinity ligand and of separating the released first analyte by at least one mode of capillary electrophoresis.

56. The electrophoresis apparatus of claim 1 wherein the first immobilized affinity ligand is adapted to bind to a corresponding affinity target for concentration.

57. The electrophoresis apparatus of claim 1 wherein the first immobilized affinity ligand is adapted to bind to a corresponding affinity target for a chemical or biochemical microreaction.

58. The electrophoresis apparatus of claim 1 further comprising an analyte concentrator containing at least one immobilized specific or non-specific affinity ligand in the transport capillary or channel and upstream of the first intersection and a valve operatively between the analyte concentrator and the first intersection.

59. The electrophoresis apparatus of claim 1 wherein the analyte detector means includes one or more first analyte detectors for the first separation capillary or channel and one or more second analyte detectors for the second separation capillary or channel.

60. The electrophoresis apparatus of claim 1 wherein at least one of the separation capillaries or channels transports analytes tagged with a chromophoric agent to the analyte detector means.

61. The electrophoresis apparatus of claim 1 wherein the first elongated concentration area is aligned with the first separation capillary or channel and is substantially perpendicular to the transport capillary or channel.

62. The electrophoresis apparatus of claim 1 wherein the first elongated concentration area is aligned with the transport capillary or channel and is substantially perpendicular to the first separation capillary or channel.

176. The electrophoresis apparatus of claim 1 further comprising a buffer supply positioned upstream of the first intersection and an analyte concentrator containing highly-specific affinity ligands and positioned upstream of the first intersection.

179. The electrophoresis apparatus of claim 1 wherein one electrode of the electrophoresis apparatus is generally at an inlet end of the separation capillaries or channels and an opposite electrode is downstream of the analyte concentrators.

63. An electrophoresis apparatus, comprising:

a transport capillary or channel;

a first analyte concentrator including one or more first immobilized affinity ligands which are attracted to a first analyte of interest;

a first separation capillary or channel to convey by electrophoresis migration and/or pressure the first analyte of interest from a sample transported in the transport capillary or channel and concentrated by the first analyte concentrator at a first location of the transport capillary or channel to a detector system which identifies and characterizes the first analyte of interest;

the transport capillary or channel and the first separation capillary or channel defining a first staggered configuration at the first location and having a first elongated section in which the first analyte concentrator is positioned;

the first separation capillary or channel being communicable upstream of the first staggered configuration with a buffer supply;

a second analyte concentrator including one or more second immobilized affinity ligands which are attracted to a second analyte of interest;

a second separation capillary or channel to convey by electrophoresis migration and/or pressure the second analyte of interest from the sample transported in the transport capillary or channel and concentrated by the second analyte concentrator at a second location of the transport capillary or channel to the detector system which also identifies and characterizes the second analyte of interest;

the transport capillary or channel and the second separation capillary or channel defining a second staggered configuration at the second location and having a second elongated section in which the second analyte concentrator is positioned;

the second separation capillary or channel being communicable upstream of the second staggered configuration with a buffer supply; and

a valve system to control fluid flow in the transport capillary or channel and the separation capillaries or channels.

64. The electrophoresis apparatus of claim 63 wherein the sample is transported by at least one of electrophoretic migration, pressure and vacuum in the transport capillary or channel.

65. The electrophoresis apparatus of claim 63 wherein the first analyte concentrator includes immobilized affinity ligands which are attached covalently to the inner wall of the first elongated section for attracting an affinity target.

66. The electrophoresis apparatus of claim 63 wherein the first analyte concentrator includes immobilized affinity ligands which are attached covalently to polymeric materials or beads located within the first elongated section for attracting an affinity target.

67. The electrophoresis apparatus of claim 63 wherein the first immobilized affinity ligands bind a complementary affinity target.

68. The electrophoresis apparatus of claim 63 wherein the first analyte concentrator includes a matrix assembly having a surface to which the first immobilized affinity ligands are bound.

69. The electrophoresis apparatus of claim 68 wherein the matrix assembly includes a plurality of microstructures.

70. The electrophoresis apparatus of claim 68 wherein the matrix assembly is free-floating and the first analyte concentrator retains the free-floating matrix assembly by pressure-resistant porous end walls disposed in the transport capillary or channel and the first separation capillary or channel.

71. The electrophoresis apparatus of claim 68 wherein the matrix assembly includes a fixed architecture defined by beaded microstructures interconnected to each other and to an inner wall of the first elongated section.

72. The electrophoresis apparatus of claim 68 wherein the matrix assembly includes a fixed architecture fabricated from polymeric microstructures interconnected to each other and to the first elongated section.

73. The electrophoresis apparatus of claim 63 further comprising an auxiliary capillary through which a cleaning buffer and a separation buffer can be introduced into the first separation capillary or channel downstream of the first analyte concentrator.

74. The electrophoresis apparatus of claim 73 wherein the auxiliary capillary is an electrolyte-provider and cleaning solution-provider capillary.

75. The electrophoresis apparatus of claim 63 further comprising an auxiliary capillary coupled to the second separation capillary or channel downstream of the second analyte capillary concentrator to provide a fluid to the second separation capillary or channel away from the second analyte concentrator.

76. The electrophoresis apparatus of claim 75 wherein flow in the auxiliary capillary is controlled by the valve system.

77. The electrophoresis apparatus of claim 63 further comprising an auxiliary analyte concentrator on the first separation capillary or channel and downstream of the first analyte concentrator, the auxiliary analyte concentrator having affinity ligands capable of retaining chromophores to bind the first analyte of interest released from the first analyte concentrator to improve the sensitivity and selectivity of the first analyte of interest.

78. The electrophoresis apparatus of claim 63 wherein the first separation capillary or channel is filled with an electrically conductive fluid.

79. The electrophoresis apparatus of claim 63 wherein the first separation capillary or channel is filled with a gel matrix and an electrically conductive fluid.

80. The electrophoresis apparatus of claim 63 wherein the sample has a plurality of proteins with different isoelectric point levels, which are further separated through the separation capillary or channel by at least one mode of capillary electrophoresis after being subjected to isoelectric focusing in the transport capillary or channel.

81. The electrophoresis apparatus of claim 63 wherein the first and/or second immobilized affinity ligands are capable of performing at least one chemical or biochemical reaction.

82. The electrophoresis apparatus of claim 80 wherein the reaction includes peptide synthesis, nucleic acid synthesis or an enzymatic reaction.

83. The electrophoresis apparatus of claim 63 wherein at least one of the analyte concentrators has an encapsulated cellular or subcellular structure adapted for drug metabolism studies.

84. The electrophoresis apparatus of claim 63 wherein at least one of the analyte concentrators has an acoustic micromixing system.

85. The electrophoresis apparatus of claim 63 wherein at least one of the analyte concentrators has a microwave pulse system.

86. The electrophoresis apparatus of claim 63 wherein the first immobilized affinity ligands are covalently bound to a matrix assembly of the first analyte concentrator.

87. The electrophoresis apparatus of claim 63 further comprising separation buffer fluid means for providing a separation buffer fluid to the first separation capillary or channel and downstream of the first analyte concentrator and to the second separation capillary or channel and downstream of the second analyte concentrator.

88. The electrophoresis apparatus of claim 87 wherein the valve system controls the operation of the separation buffer fluid means.

89. The electrophoresis apparatus of claim 87 wherein the separation buffer fluid includes an organic solvent, or a mixture of organic solvents and additives.

90. The electrophoresis apparatus of claim 87 wherein the separation buffer fluid contains sodium tetraborate.

91. The electrophoresis apparatus of claim 63 further comprising a first electrolyte-provider capillary or channel in fluid communication with the first separation capillary or channel and downstream of the first analyte concentrator and a second electrolyte-provider capillary or channel in fluid communication with the second separation capillary or channel and downstream of the second analyte concentrator.

92. The electrophoresis apparatus of claim 63 wherein the detector system is an ultraviolet detector system.

93. The electrophoresis apparatus of claim 63 wherein the detector system is a fluorescence or laser-induced fluorescence detector system.

94. The electrophoresis apparatus of claim 63 wherein the detector system is a conductivity, electrochemical, radioactive, mass spectrometer, circular diochroism or nuclear magnetic resonance detector system or a combination of several detectors used simultaneously.

95. The electrophoresis apparatus of claim 63 wherein the first and second separation capillaries or channels merge into a single exit outlet capillary or channel.

96. The electrophoresis apparatus of claim 95 wherein the valve system controls, at the merging of the first and second separation capillaries or channels, sequential fluid flow from the first and second separation capillaries or channels to the exit outlet capillary or channel.

97. The electrophoresis apparatus of claim 95 further comprising a grounding electrode at an outlet end of the exit outlet capillary or channel.

98. The electrophoresis apparatus of claim 97 wherein the exit outlet capillary or channel flows into a container in which the grounding electrode is positioned.

99. The electrophoresis apparatus of claim 63 further comprising an exit outlet capillary or channel into which the first and second separation capillaries or channels flow and at a detection zone of the detector system.

100. The electrophoresis apparatus of claim 99 wherein the valve system controls sequential fluid flow of the first and second separation capillaries or channels to the exit output capillary or channel and the detection zone.

101. The electrophoresis apparatus of claim 63 further comprising capillary or channel bulging members which retain the first immobilized affinity ligands in the first analyte concentrator.

102. The electrophoresis apparatus of claim 63 wherein the inner diameter of the transport capillary or channel is larger than the inner diameter of the first separation capillary or channel and than the inner diameter of the second separation capillary or channel.

103. The electrophoresis apparatus of claim 63 wherein the transport capillary or channel and the first and second separation capillaries or channels are all capillaries.

104. The electrophoresis apparatus of claim 63 wherein the transport capillary or channel and the first and second separation capillaries or channels are all channels.

105. The electrophoresis apparatus of claim 63 wherein the electrophoresis apparatus is a capillary electrophoresis apparatus.

106. The electrophoresis apparatus of claim 63 wherein the electrophoresis apparatus is a microchip electrophoresis apparatus.

107. The electrophoresis apparatus of claim 63 wherein the first immobilized affinity ligands are covalently linked to an inner wall of the first elongated section.

108. The electrophoresis apparatus of claim 63 wherein the valve system includes transport capillary or channel valves and separation capillary or channel valves, and wherein the transport capillary or channel valves are adapted to be opened and the first separation capillary or channel valves are adapted to be closed to allow fluid to pass through the first analyte concentrator towards an outlet end of the transport capillary or channel.

109. The electrophoresis apparatus of claim 108 wherein the fluid is the sample, at least one chromophoric substance, and at least one cleaning buffer.

110. The electrophoresis apparatus of claim 63 wherein the valve system includes transport capillary or channel valves and first separation capillary or channel valves, and wherein the transport capillary or channel valves are adapted to be closed and the first separation capillary or channel valves are adapted to be opened to allow a separation buffer solution to pass through the first analyte concentrator and in the first separation capillary or channel to the detector system.

111. The electrophoresis apparatus of claim 63 wherein the positive electrode of the electrophoresis apparatus is at the buffer supply and the negative electrode is downstream of the analyte concentrators.

112. The electrophoresis apparatus of claim 63 wherein an inlet end of the separation capillary or channel is alternatively in fluid communication with a cleaning solution supply or an eluting buffer supply or a separation buffer supply.

113. The electrophoresis apparatus of claim 63 wherein the capillaries or channels are fused-silica or plastic tubes or channels.

114. The electrophoresis apparatus of claim 63 wherein the buffer supply of the first separation capillary or channel includes a separation buffer and an eluting buffer to release the bound first analyte of interest from the first immobilized affinity ligands.

115. The electrophoresis apparatus of claim 63 wherein the valve system include valves on the transport capillary or channel on opposite sides of the first analyte concentrator and valves on the separation capillary or channel on opposite sides of the first analyte concentrator.

116. The electrophoresis apparatus of claim 63 wherein the first separation capillary or channel is positioned and capable of separating therein the first analyte of interest retained by the first immobilized affinity ligands after the first analyte is released from the first immobilized affinity ligands and of separating the released first analyte by at least one mode of capillary electrophoresis.

117. The electrophoresis apparatus of claim 63 further comprising an analyte concentrator containing at least one specific or non-specific analyte positioned upstream of the first intersection.

118. The electrophoresis apparatus of claim 63 wherein the at least one first immobilized affinity ligands is oriented to facilitate binding between the first immobilized affinity ligands and an affinity target.

119. The electrophoresis apparatus of claim 63 wherein the first immobilized affinity ligands are oriented in a direction to increase surface area to increase capacity to capture an affinity target.

177. The electrophoresis apparatus of claim 63 further comprising a buffer supply positioned upstream of the first intersection and an analyte concentrator containing highly-specific affinity ligands and positioned upstream of the first intersection.

180. The electrophoresis apparatus of claim 63 wherein one electrode of the electrophoresis apparatus is generally at an inlet end of the separation capillaries or channels and an opposite electrode is downstream of the analyte concentrators.

120. An electrophoresis apparatus, comprising:

a transport capillary or channel;

a first analyte concentrator which is a first analyte concentrator-microreactor adapted to concentrate a first analyte of interest;

a first separation capillary or channel to convey by electrophoresis migration and/or pressure the first analyte of interest from a sample transported in the transport capillary or channel and concentrated by one or more first immobilized affinity ligands in the first analyte concentrator-microreactor at a first location of the transport capillary or channel to a detector system which identifies and characterizes the first analyte of interest;

the first separation capillary or channel being positionable in fluid communication at an inlet end thereof with a buffer supply;

the transport capillary or channel and the first separation capillary or channel defining a first staggered configuration at the first location and having a first elongated portion in which the first analyte concentrator-microreactor is positioned;

a second analyte concentrator which is a second analyte concentrator-microreactor adapted to concentrate a second analyte of interest;

a second separation capillary or channel to convey by electrophoresis migration and/or pressure the second analyte of interest from the sample and concentrated by one or more second immobilized affinity ligands in the second analyte concentrator-microreactor at a second location of the transport capillary or channel to the detector system which also identifies and characterizes the second analyte of interest;

the second separation capillary or channel being positionable in fluid communication at an inlet end thereof with a buffer supply;

the transport capillary or channel and the second separation capillary or channel defining a second staggered configuration at the second location and having a second elongated portion in which the second analyte concentrator-microreactor is positioned; and

a valve system to control fluid flow in the capillaries or channels, the valve system including valves on the first separation capillary or channel and the transport capillary or channel and operatively around the first staggered configuration and valves on the second separation capillary or channel and the transport capillary or channel and operatively around the second staggered configuration.

121. The electrophoresis apparatus of claim 120 wherein the sample is transported by electrophoretic migration, pressure and/or vacuum into the transport capillary or channel.

122. The electrophoresis apparatus of claim 120 wherein the first analyte concentrator-microreactor comprises a site for chemical synthesis.

123. The electrophoresis apparatus of claim 122 wherein the chemical synthesis is peptide synthesis or nucleic acid synthesis.

124. The electrophoresis apparatus of claim 120 wherein the first concentrator-microreactor includes a matrix assembly having a surface to which the first immobilized affinity ligands are bound.

125. The electrophoresis apparatus of claim 124 wherein the matrix assembly includes a plurality of microstructures.

126. The electrophoresis apparatus of claim 124 wherein the matrix assembly is free-floating and the first concentrator-microreactor retains the free-floating matrix assembly by pressure-resistant porous end walls or frits disposed in the transport capillary or channel and the first separation capillary or channel.

127. The electrophoresis apparatus of claim 124 wherein the matrix assembly includes a fixed architecture defined by beaded microstructures interconnected to each other and to an inner wall of the first separation capillary or channel.

128. The electrophoresis apparatus of claim 124 wherein the matrix assembly includes a fixed architecture fabricated from polymeric microstructures interconnected to each other and to an inner wall of the first elongated portion.

129. The electrophoresis apparatus of claim 120 further comprising an auxiliary capillary through which a cleaning buffer and a separation buffer can be introduced into the first separation capillary or channel and downstream of the first analyte concentrator-microreactor.

130. The electrophoresis apparatus of claim 129 wherein the auxiliary capillary is an electrolyte-provider and a cleaning solution-provider capillary.

131. The electrophoresis apparatus of claim 120 further comprising an auxiliary capillary coupled to the first separation capillary or channel downstream of the first analyte concentrator-microreactor to provide a fluid to the first separation capillary or channel away from the first analyte concentrator-micro reactor.

132. The electrophoresis apparatus of claim 131 wherein the auxiliary capillary is controlled by the valve system.

133. The electrophoresis apparatus of claim 120 further comprising an auxiliary analyte concentrator on the first separation capillary or channel and downstream of the first analyte concentrator-microreactor, the auxiliary analyte concentrator having affinity ligands capable of retaining chromophores to bind the first analyte of interest released from the first analyte concentrator-microreactor to improve the sensitivity and selectivity of the first analyte of interest.

134. The electrophoresis apparatus of claim 120 wherein the first separation capillary or channel is filled with an electrically conductive fluid.

135. The electrophoresis apparatus of claim 120 wherein the first separation capillary or channel is filled with a gel matrix and an electrically conductive fluid.

136. The electrophoresis apparatus of claim 120 wherein the sample has a plurality of proteins with different isoelectric point levels, which are further separated through the separation capillaries or channels by at least one mode of capillary electrophoresis after being subjected to isoelectric focusing in the transport capillary or channel.

137. The electrophoresis apparatus of claim 120 wherein the first and/or second immobilized affinity ligands are capable of performing at least one chemical or biochemical reaction.

138. The electrophoresis apparatus of claim 137 wherein the reaction is peptide synthesis, nucleic acid synthesis, or an enzymatic reaction.

139. The electrophoresis apparatus of claim 120 wherein the first and/or second analyte concentrator-microreactors have an encapsulated cellular or subcellular structure.

140. The electrophoresis apparatus of claim 139 wherein the encapsulated cellular or subcellular structure is adapted for drug metabolism studies and/or for metabolic pathway studies.

141. The electrophoresis apparatus of claim 120 wherein the first and/or second analyte concentrator-microreactors have an acoustic micromixing system.

142. The electrophoresis apparatus of claim 120 wherein the first and/or second analyte concentrator-microreactors have a microwave pulse system.

143. The electrophoresis apparatus of claim 120 wherein first immobilized affinity ligands are covalently bound to a matrix assembly of the first concentrator-microreactor.

144. The electrophoresis apparatus of claim 120 further comprising separation buffer fluid means for providing a separation buffer fluid to the first separation capillary or channel and downstream of the first analyte concentrator-microreactor and to the second separation capillary or channel and downstream of the second analyte concentrator-microreactor.

145. The electrophoresis apparatus of claim 144 wherein the valve system controls the operation of the separation buffer fluid means.

146. The electrophoresis apparatus of claim 144 wherein the separation buffer fluid includes an organic solvent, or a mixture of organic solvents or additives.

147. The electrophoresis apparatus of claim 144 wherein the separation buffer fluid contains sodium tetraborate.

148. The electrophoresis apparatus of claim 120 further comprising a first electrolyte-provider capillary or channel in fluid communication with the first separation capillary or channel and downstream of the first analyte concentrator-microreactor and a second electrolyte-provider capillary or channel in fluid communication with the second separation capillary or channel and downstream of the second analyte concentrator-microreactor.

149. The electrophoresis apparatus of claim 120 wherein the detector system is an ultraviolet detector system.

150. The electrophoresis apparatus of claim 120 wherein the detector system is a fluorescence or laser-induced fluorescence detector system.

151. The electrophoresis apparatus of claim 120 wherein the detector system is a conductivity, electrochemical, radioactive, mass spectrometer,

circular diochroism or nuclear magnetic resonance detector system.

152. The electrophoresis apparatus of claim 120 wherein the first and second separation capillaries or channels merge into a single exit output capillary or channel.

153. The electrophoresis apparatus of claim 152 wherein the valve system controls, at the merging of the first and second separation capillaries or channels, sequential fluid flow from the first and second separation capillaries or channels to the exit output capillary or channel.

154. The electrophoresis apparatus of claim 120 further comprising an exit outlet capillary or channel into which the first and second separation capillaries or channels flow and at a detection zone of the detector system.

155. The electrophoresis apparatus of claim 154 wherein the valve system controls sequential fluid flow of the first and second separation capillaries or channels to the detection zone.

156. The electrophoresis apparatus of claim 154 further comprising a grounding electrode at an outlet end of the exit outlet capillary or channel.

157. The electrophoresis apparatus of claim 156 wherein the exit outlet capillary or channel flows into a container in which the grounding electrode is positioned.

158. The electrophoresis apparatus of claim 120 further comprising capillary or channel bulging members which retain the first immobilized affinity ligands in the first analyte concentrator-microreactor.

159. The electrophoresis apparatus of claim 120 wherein the inner diameter of the transport capillary or channel is larger than the inner diameter of the first separation capillary or channel and than the inner diameter of the second separation capillary or channel.

160. The electrophoresis apparatus of claim 120 wherein the transport capillary or channel and the first and second separation capillaries or channels are all capillaries.

161. The electrophoresis apparatus of claim 120 wherein the transport capillary or channel and the first and second separation capillaries or channels are all channels.

162. The electrophoresis apparatus of claim 120 wherein the electrophoresis apparatus is a capillary electrophoresis apparatus.

163. The electrophoresis apparatus of claim 120 wherein the electrophoresis apparatus is a microchip electrophoresis apparatus.

164. The electrophoresis apparatus of claim 120 wherein first immobilized affinity ligands of the first analyte concentrator-microreactor are covalently linked to an inner wall of the first elongated portion.

165. The electrophoresis apparatus of claim 120 wherein the valve system includes transport capillary or channel valves and separation capillary or channel valves, and wherein the transport capillary or channel valves are adapted to be opened and the first separation capillary or channel valves are adapted to be closed to allow fluid to pass through the first concentrator-microreactor towards an outlet end of the transport capillary or channel.

166. The electrophoresis apparatus of claim 165 wherein the fluid is the sample, at least one chromophoric substance, and at least one cleaning buffer.

167. The electrophoresis apparatus of claim 120 wherein the valve system includes transport capillary or channel valves and first separation capillary or channel valves, and wherein the transport capillary or channel valves are adapted to be closed and the first separation capillary or channel valves are adapted to be opened to allow a separation buffer solution to pass through the first analyte concentrator-microreactor and in the first separation capillary or channel to the detector system.

168. The electrophoresis apparatus of claim 120 wherein the positive electrode of the electrophoresis apparatus is generally at the buffer supply and the negative electrode is downstream of the analyte concentrator-microreactors.

169. The electrophoresis apparatus of claim 120 wherein the first separation capillary or channel is a first separation capillary and an inlet end of the first separation capillary is alternatively in fluid communication with a cleaning solution supply or an eluting buffer supply or a separation buffer supply.

170. The electrophoresis apparatus of claim 120 wherein the capillaries or channels are fused-silica or plastic tubes or channels.

171. The electrophoresis apparatus of claim 120 wherein the buffer supply of the first separation capillary or channel includes a separation buffer and an eluting buffer to release the bound first analyte of interest from first immobilized affinity ligands of the first analyte concentrator-microreactor.

172. The electrophoresis apparatus of claim 120 wherein the first separation capillary or channel is positioned and capable of separating therein the first analyte of interest retained by the first immobilized affinity ligands after the first analyte of interest is released from the first immobilized affinity ligands and of separating the released analyte by at least one mode of capillary electrophoresis.

173. The electrophoresis apparatus of claim 120 further comprising a buffer supply positioned upstream of the first intersection and an analyte concentrator containing non-specific affinity ligands and positioned upstream of the first location.

174. The electrophoresis apparatus of claim 120 further comprising a buffer supply positioned upstream of the first intersection and an analyte concentrator containing highly-specific affinity ligands and positioned upstream of the first location.

175. The electrophoresis apparatus of claim 120 wherein at least one of the separation capillaries or channels transports analytes tagged with a chromophoric agent to the detector system.

181. The electrophoresis apparatus of claim 120 wherein one electrode of the electrophoresis apparatus is generally at an inlet end of the separation capillaries or channels and an opposite electrode is downstream of the analyte concentrator-microreactors.

178. An electrophoresis apparatus, comprising:

a transport capillary or channel;

a first separation capillary or channel overlapping and intersecting the transport capillary or channel at a first intersection;

the first intersection having a first staggered configuration which includes a first elongated concentration area;

a first analyte concentrator containing at least one first immobilized affinity ligand in the first elongated concentration area to concentrate a first analyte of interest from a sample introduced into the transport capillary or channel;

a second separation capillary or channel overlapping and intersecting the transport capillary or channel at a second intersection spaced downstream from the first intersection;

the second intersection having a second staggered configuration which includes a second elongated concentration area;

a second analyte concentrator containing at least one second immobilized affinity ligand in the second elongated concentration area to concentrate a second analyte of interest from the sample introduced into the transport capillary or channel;

analyte detector means for identifying and characterizing the first and second analytes of interest conveyed thereto from the first and second analyte concentrators, respectively; and

a valve system which controls flow of the sample in the transport capillary or channel and past the first and second intersections and which controls flow of buffer fluid through the first and second separation capillaries or channels and conveyed by electrophoresis migration, pressure or a combination of electrophoresis migration and pressure to the analyte detector means.

182. The electrophoresis apparatus of claim 178 wherein one electrode of the electrophoresis apparatus is generally at an inlet end of the separation capillaries or channels and an opposite electrode is downstream of the analyte concentrators.

183. The electrophoresis apparatus of claim 178 wherein the capillaries or channels are all capillaries.

184. The electrophoresis apparatus of claim 178 wherein the capillaries or channels are all channels.