Electrophoresis apparatus having an outlet passage

• US 7,811,436 B2
• Filed: 10/31/2007
• Issued: 10/12/2010
• 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): NOGUEROLA, ALEXANDER STEPHAN
• Application Number:
  US11/980,922
• Publication Number:
  US 20080060944A1
• Time in Patent Office: 1,077 Days
• Field of Search: 204601-605, 204451-455
• US Class Current: 204/604
• CPC Class Codes: G01N 27/44704 Details; Accessories
• 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

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. Analyte concentrators, having antibody-specific (or related affinity) chemistries, are stationed at the respective inter-sections 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.

Claims

1. An electrophoresis apparatus comprising:

a transport passage having an inlet entrance and an exit outlet;

a first separation passage which intersects with the transport passage at a first intersection;

the first separation passage having an inlet entrance and an exit outlet;

a first concentrator to concentrate a first analyte;

the first concentrator being positioned at the first intersection;

a second separation passage which intersects with the transport passage at a second intersection;

the second separation passage having an inlet entrance and an exit outlet;

a second concentrator to concentrate a second analyte;

the second concentrator being positioned at the second intersection;

an outlet passage;

the first separation passage to convey to the outlet passage the first analyte concentrated by the first concentrator from a sample introduced into the transport passage;

the second separation passage to convey to the outlet passage the second analyte concentrated by the second concentrator from the sample introduced into the transport passage; and

a detector to identify and characterize the first and second analytes when delivered sequentially thereto in the outlet passage.

2. The apparatus of claim 1 wherein the detector senses ultraviolet energy.

3. The apparatus of claim 1 wherein the detector senses fluorescent energy.

4. The apparatus of claim 1 wherein each of the passages is a capillary.

5. The apparatus of claim 1 wherein each of the passages is at least substantially a channel.

6. The apparatus of claim 1 wherein the outlet passage connects to a grounding and waste container.

7. The apparatus of claim 1 wherein the inlet entrance of the first separation passage is a buffer solution inlet end, and the inlet entrance of the second separation passage is a buffer solution inlet end.

8. The apparatus of claim 7 further comprising a first buffer supply in direct fluid communication with the buffer solution inlet end of the first separation passage and a second buffer supply in direct fluid communication with the buffer solution inlet end of the second separation passage.

9. The apparatus of claim 1 further comprising:

a third concentrator to concentrate a third analyte;

a third separation passage to convey to the outlet passage the third analyte concentrated by the third concentrator from the sample;

the third concentrator being positioned at an intersection of the third separation passage and the transport passage; and

the detector identifying and characterizing the first, second and third analytes when delivered sequentially thereto in the outlet passage.

10. The apparatus of claim 9 wherein all of the passages are capillaries.

11. The apparatus of claim 1 wherein the second intersection is spaced a distance downstream of the first intersection.

12. The apparatus of claim 1 wherein the first and second concentrators are both affinity concentrators.

13. The apparatus of claim 1 wherein the first concentrator includes microstructures carrying at least one affinity element adapted to attract the first analyte from the sample passing through the first concentrator, and the second concentrator includes microstructures carrying at least one affinity element adapted to attract the second analyte from the sample passing through the second concentrator.

14. The apparatus of claim 1 wherein the first concentrator has a matrix assembly provided by a plurality of microstructures and porous end walls disposed in the transport passage and the first separation passage to retain the microstructures.

15. The apparatus of claim 1 wherein the detector includes an on-column detector, and further comprising an off-column detector downstream from the on-column detector.

16. The apparatus of claim 15 wherein the on-column detector senses ultraviolet or fluorescent energy.

17. The apparatus of claim 16 wherein the off-column detector comprises an electrochemical detector, a mass spectrometry detector, a circular dichroism detector or a nuclear magnetic resonance detector.

18. The apparatus of claim 1 wherein the transport passage has an inner diameter larger than the inner diameters of the first and second separation passages.

19. The apparatus of claim 1 wherein all of the transport and separation passages are formed into a microchip.

20. The apparatus of claim 1 wherein the first and second separation passages are both perpendicular to the transport passage.

21. The apparatus of claim 1 wherein the first and second analytes are conveyed at least by electrophoresis migration in the outlet passage and past the detector.

22. The apparatus of claim 1 further comprising an auxiliary analyte concentrator in the first separation passage between the first concentrator and the outlet passage.

23. The apparatus of claim 22 wherein the auxiliary analyte concentrator includes microstructures coated with:

(1) a chromophoric agent which increases ultraviolet absorptivity, fluorescence or phosphorescence when bound to a minute quantity of the first analyte when forming a derivatized analyte or conjugate analyte-chromophore, or

(2) a chromophoric agent which increases ultraviolet absorptivity, fluorescence or phosphorescence when bound to a minute quantity of the first analyte when forming a derivatized analyte or conjugate analyte-chromophore, and in which an antibody that is immobilized to the microstructures and with an antibody that binds to a portion of the chromophoric agent.

24. The apparatus of claim 23 further comprising a cleansing passage intersecting the first separation passage at the auxiliary analyte concentrator and carrying a buffer to cleanse the microstructures.