Electrophoresis capillary with agarose

• US 5,089,103 A
• Filed: 12/01/1989
• Issued: 02/18/1992
• Est. Priority Date: 12/01/1989
• Status: Expired due to Term

• Current Assignee: Agilent Technologies, Inc.
• Original Assignee: Hewlett-Packard Company (HP Inc.)
• Inventors: Swedberg, Sally A.
• Primary Examiner(s): Niebling, John
• Assistant Examiner(s): RYSER, DAVID
• Application Number:
  US07/444,229
• Time in Patent Office: 809 Days
• Field of Search: 204/299 R, 204/182.8
• US Class Current: 204/605
• CPC Class Codes: B01D 57/02 by electrophoresis treatment of water, waste water, sewage or sludge by electrophoresis C02F1/469; electrophoretic production of compounds or non-metals C25B7/00; investigating or analysing materials by using electrophoresis G01N27/26
• CPC Class Codes: G01N 27/44704 Details; Accessories
• CPC Class Codes: G01N 27/44747 Composition of gel or of carrier mixture

Abstract

Capillary tubes useful for performing capillary zone electrophoresis separation techniques are prepared by precoating the capillary tubes with an adhesive agent, preferably a dilute agarose phase. The analytical agarose-based media then is positioned in the tubes, where it adheres to the earlier coating. The analytical media exhibits strong structural integrity and adheres well to the capillary tubes; moreover, the media is suitable for capillary zone electrophoretic separation of biopolymers.

Claims

1. A method for preparing a narrow-bore capillary tube for use in electrophoretic separations comprising the steps of:

contacting a dilute agarose phase wit the bore of the tube to for m a precoat thereon; and

adherently filling the bore with an agarose-based analytical gel along at least a portion of the tube which is precoated with the dilute agarose phase.

2. The method as defined in claim 1 wherein the analytical gel is adapted for separation of biopolymers substantially based on charge differences.

3. The method as defined in claim 2 wherein the analytical gel has an agarose concentration from about 0.5 wt. % to about 5 wt. %.

4. The method as defined in claim 3 wherein the gel is adapted to separate biopolymer solutes by molecular sieving.

5. An apparatus useful for electrophoretic separations of bipolymer solutes comprising:

a capillary tube defining a bore surface;

a precoat adherently coating at least a portion of the bore surface, the precoat including an agarose phase; and

an analytical agarose-based gel adhered in the capillary, which gel is adherent to the precoat and is permeable to biopolymer solutes moving in response to sufficient electric current for separation of the biopolymers.

6. The apparatus as defined in claim 5 wherein the analytical gel fills the capillary as a continuous phase, and the analytical gel is adapted to maintain continuity when subjected to an applied field of between about 300 V/cm to about 400 V/cm tube length.

9. The apparatus as defined in claim 5 wherein the analytical gel is adapted for separation of bipolymers substantially based on charge differences.

10. An apparatus useful for electrophoretic separations of biopolymer solutes comprising:

a capillary tube having a bore size between about 75 μ

to about 500 μ and

defining a bore surface; and

,

an agarose-based media being permeable to biopolymer solutes and disposed within the bore as a continuous gelled phase, the agarose-based media radially outwardly extending to the bore surface and adhered thereto and longitudinally extending along sufficient of the tube length to separate biopolymers substantially on the basis of charge differences, the agarose-based media being sufficiently stable as to maintain continuity when subjected to an electric field of about 300 V/cm tube length.

7. The apparatus as defined in claim 12 wherein the analytical gel has an agarose concentration from about 0.5 wt. % to about 5 wt. %.

8. The apparatus as defined in claim 7 wherein the gel is adapted to separate biopolymer solutes by molecular sieving.

11. The apparatus of claim 10 wherein the agarose-based media includes agarose in an amount from about 0.5 wt. % to about 5 wt. %, and further includes an additive effective to increase mechanical stability in an amount from about 0.5% to about 10%, the agarose-based media being adhered to the bore surface by an adhesive agent.

12. The apparatus of claim 11 wherein the adhesive agent is effective to maintain the media adhered to the bore surface during separations of biopolymers by adhesive forces, and the adhesive agent includes agarose.