Electrophoretic separating device and electrophoretic separating method

• US 5,296,114 A
• Filed: 11/30/1992
• Issued: 03/22/1994
• Est. Priority Date: 12/06/1991
• Status: Expired due to Term

• Current Assignee: Zeptosens AG (Bayer AG)
• Original Assignee: Ciba-Geigy Corporation (Novartis Ag)
• Inventors: Manz, Andreas
• Primary Examiner(s): Niebling, John
• Assistant Examiner(s): STARSIAK, JOHN S
• Application Number:
  US07/983,178
• Time in Patent Office: 477 Days
• Field of Search: 204/299 R, 204/180.1
• US Class Current: 204/451
• CPC Class Codes: G01N 27/44704 Details; Accessories
• CPC Class Codes: G01N 27/44773 Multi-stage electrophoresis, e.g. two-dimensional electrophoresis

Abstract

In an electrophoretic separating device, a channel defining a separating path is constructed in the form of a closed loop. A sample to be separated is introduced via a feed opening into an electrolytic carrier medium that is moved with the aid of an electric field through the channel, which is provided in the region of its ends with inlet and outlet openings for the carrier medium, and is separated into individual components by the electric field. The electric field is generated in the channel by connecting electrodes in the region of the inlet and outlet openings to different potentials of a voltage source. The procedure for carrying out the method according to the invention is distinguished especially by the fact that the carrier medium and the sample are moved along the substantially closed separating path formed by the channel which is constructed in the form of a closed loop.

Claims

1. An electrophoretic separating device comprising a channel which is provided with inlet and outlet openings, for example for a carrier medium, and which has at least one feed opening for a sample to be investigated, and having electrodes in the region of the inlet and outlet openings, which electrodes can be connected to a voltage source in such a manner than an electric field can be established along the channel, wherein the channel is divided into n channel sections and has, in addition to the feed opening for the sample, 2n inlet and outlet openings that are arranged in the region of connection between the channel sections, and wherein the channel is constructed in the form of an n-sided closed loop figure, the sides of which are formed by the channel sections.

2. A device according to claim 1, wherein the channel is constructed in the manner of a capillary.

3. A device according to claim 1, wherein each channel section is associated with electrodes which can be connected to the voltage source in such a manner that the electric field can be applied substantially along a channel region that is arranged between the relevant electrodes, which electrodes have different electrical potentials.

4. A device according to claim 3, wherein the channel region along which the electric field is applied comprises at least two channel sections the connection region of which is associated with an electrode having a potential lying between the potentials of the source electrodes of the electric field.

5. A device according to claim 4, wherein the voltage source is a direct voltage source.

6. A device according to claim 5, wherein the electrodes can be connected to the voltage source in pairs, preferably with the aid of a relay, so as to generate an electric field that migrates cyclically along the individual channel regions.

7. A device according to claim 1, wherein the channel comprises at least three channel sections.

8. A device according to claim 1, which is constructed in the form of a miniaturised component which is preferably of two-part construction and comprises a body part and a lid part, preferably of glass, with the channel, the inlet and outlet openings and the feed opening preferably being provided on the body part.

9. A device according to claim 8, wherein the body part is a micromechanically or photolithographically machined glass plate.

10. A device according to claim 8, wherein the body part is a preferably photolithographically or micromechanically machined plate of semi-conductive material, preferably a silicon plate.

11. A device according to claim 1, which comprises at least one detector, for example an absorption detector for ultraviolet radiation or for the visible spectrum, or a luminescence detector or an electrochemical detector, which is arranged along the channel.

12. A device according to claim 1, wherein the channel has a cross-sectional surface area of from approximately 100 μ

m² to approximately 5000 μ

m².

13. An electrophoretic separating method wherein a sample is introduced via a feed opening into an electrolytic carrier medium that is moved with the aid of an electric field through a channel being provided with inlet and outlet openings for the carrier medium, and is separated into individual components by the electric field which is generated in the channel by connecting electrodes in the region of the inlet and outlet openings to different potentials of a voltage source, which method comprises moving the carrier medium and the sample along a substantially closed path formed by the channel which is constructed in the form of an n-sided closed loop figure and that is provided with 2n inlet and outlet openings in addition to the feed opening for the sample.

14. A method according to claim 13, wherein the carrier medium and the sample are moved repeatedly, especially cyclically, through the channel.

15. A method according to claim 13, wherein the electrical potential difference for generating the electric field for transporting the carrier medium and the sample is generated essentially only in a relevant portion of the channel, especially in that portion in which the sample is located, and, in accordance with the desired transport direction of the carrier medium and of the sample or the component of interest, the field is generated in successive portions.

16. A method according to claim 15, wherein the switching frequency of the electrical potential difference to the adjacent channel regions is matched to the circulation speed of the component of interest in the channel.

17. A method according to claim 16, wherein the components that migrate more quickly or more slowly than the switching frequency are discharged through the outlet openings arranged along the channel.

18. A method according to claim 16, wherein the carrier medium and the sample or the component of interest in each channel region are moved through at least two channel sections which each comprise at least one inlet and outlet opening and also comprise electrodes which are provided in the region of each opening and are connected to a voltage source.

19. A method according to claim 18, wherein the electrodes provided at the beginning and end points of each channel region for generating the electric field are connected in pairs to a direct voltage source and the electrode(s) located therebetween is(are) physically separated from the voltage source.

20. A method according to claim 13, wherein the electrical potential difference is switched on to the adjacent channel region only after the sample has been moved into the first channel section of the adjacent channel region.

21. A method according to claim 13, wherein the channel is constructed in the manner of a capillary and with a cross-sectional surface area of from approximately 100 μ

m² to approximately 5000 μ

m², and the strength of the electric field is matched to the dimensions of the channel in such a manner that at a theoretical channel length of 1 m, the thermal heat output does not exceed 1 W.

22. A method according to claim 21, wherein the potential difference selected for generating the electric field is from approximately 100 V to several kilovolts, in the case of a semi-conductive material preferably up to about 500 V.

23. A method according to claim 13, wherein the state of separation of the sample into the individual components is examined at least once per circuit in the channel with the aid of one or more detectors arranged along the channel.