Pulsed field capillary electrophoresis

• US 5,122,248 A
• Filed: 05/18/1990
• Issued: 06/16/1992
• Est. Priority Date: 05/18/1990
• Status: Expired due to Fees

• Current Assignee: Northeastern University
• Original Assignee: Northeastern University
• Inventors: Heiger, David N., Karger, Barry L., Cohen, Aharon S.
• Primary Examiner(s): Niebling, John
• Assistant Examiner(s): Koestner, Caroline
• Application Number:
  US07/525,532
• Time in Patent Office: 760 Days
• Field of Search: 204/299 R, 204/183.3, 204/182.8
• US Class Current: 204/453
• CPC Class Codes: G01N 27/44773 Multi-stage electrophoresis, e.g. two-dimensional electrophoresis

Abstract

Apparatus and method for conducting electrophoresis in a capillary tube under the influence of a pulsed electric field. Apparatus for separating and detecting molecular species of a sample in a conductive medium includes a capillary tube for containing conductive medium, means for introducing a sample into a conductive medium-filled capillary tube, means for applying a pulsed electric field across the conductive medium-filled capillary tube between its ends, and means for detecting constituent molecular species of a sample in a conductive medium-filled capillary tube as they traverse the tube. A method for separating and detecting molecular species of a sample in a conductive medium includes the steps of introducing the sample into a capillary tube filled with conductive medium, applying a pulsed electric field of up to 500 volts/centimeter between the ends of the capillary, and detecting the constituent molecular species of the sample as they traverse the capillary.

Claims

1. Apparatus for separating and detecting a plurality of molecular species of a sample in conductive medium, comprising:

a capillary tube containing a conductive medium including a sieving matrix, said tube having a cross-sectional dimension of not more than 500 micrometers;

means for introducing said sample into said conductive medium-filled capillary tube;

means for applying a pulsed electric field across said conductive medium-filled capillary tube, between the ends thereof;

said pulsed electric field enhancing the separation of said plurality of molecular species of said sample as said sample migrates through said sieving matrix contained within said capillary tube; and

means for detecting said plurality of molecular species of said sample as said plurality of molecular species migrate within said capillary tube.

2. The apparatus of claim 1, wherein said sieving matrix comprises a polymeric material.

3. The apparatus of claim 2, wherein said polymeric material is crosslinked.

4. The apparatus of claim 1, wherein said means for introducing a sample into a capillary tube comprises an automatic sample handling device.

5. The apparatus of claim 1, wherein said means for applying a pulsed electric field comprises a waveform generator.

6. The apparatus of claim 5, wherein said means for applying a pulsed electric field further comprises an amplifier, the input of said amplifier being electrically connected to the output of said waveform generator, and the output of said amplifier being electrically connected to the respective ends of said capillary tube.

7. The apparatus of claim 6, wherein said means for applying a pulsed electric field further comprises a programmable computer having a communication port which is connected to the input of said arbitrary waveform generator.

8. The apparatus of claim 1, wherein said means for detecting comprises a detector in communication with a translucent section of said capillary tube, said detector being sensitive to change in electromagnetic radiation resulting from transit of constituent molecular species of said sample through said capillary tube.

9. The apparatus of claim 1, wherein said means for detecting comprises a mass spectrometer.

10. The apparatus of claim 1, wherein said means for detecting comprises an electrochemical detector.

11. The apparatus of claim 1, wherein said means for detecting comprises a radioactivity-sensing detector.

12. A method for separating and detecting a plurality of molecular species of a sample in conductive medium, comprising:

introducing said sample into a capillary tube having a cross-sectional dimension of not more than 500 micrometers, and filled with conductive medium including a sieving matrix;

applying a pulsed electric field of up to 500 volts per centimeter across said capillary tube, between the ends thereof;

said pulsed electric field enhancing the separation of said plurality of molecular species of said sample as said sample migrates through said sieving matrix contained within said capillary tube; and

detecting said plurality of molecular species of said sample as said plurality of molecular species migrate within said capillary tube.

13. The apparatus of claim 12, wherein the sample is introduced into the capillary tube by electrophoretic injection.

14. The apparatus of claim 12, wherein the sample is introduced by pressurized injection.

15. The apparatus of claim 12, wherein in the step of applying a pulsed electric field, the pulses are unidirectional relative to a zero field state.

16. The apparatus of claim 12, wherein in the step of applying a pulsed electric field, the applied field is positive.

17. The apparatus of claim 12, wherein in the step of applying a pulsed electric field, the pulses are bidirectional relative to a zero field state.

18. The apparatus of claim 12, wherein in the step of applying a pulsed electric field, the frequency of the pulses varies with time.

19. The apparatus of claim 12, wherein in the step of applying a pulsed electric field, the waveform of the pulses varies with time.

20. The apparatus of claim 12, wherein in the step of applying a pulsed electric field, the amplitude of successive pulses varies with time.

21. The apparatus of claim 12, wherein in the step of applying a pulsed electric field, the duty cycles varies with time.

22. The apparatus of claim 12, wherein the capillary tube includes polymeric material at a level less than 10% T.