Viscous electrophoresis polymer medium and method

• US 5,290,418 A
• Filed: 09/24/1992
• Issued: 03/01/1994
• Est. Priority Date: 09/24/1992
• Status: Expired due to Term

• Current Assignee: Applied Biosystems LLC (Thermo Fisher Scientific Incorporated)
• Original Assignee: Applied Biosystems, Inc. (Thermo Fisher Scientific Incorporated)
• Inventors: Winnik, Michell, Menchen, Steven M.
• Primary Examiner(s): Niebling, John
• Assistant Examiner(s): DELACROIX MUIRHEI, CYBILLE
• Application Number:
  US07/950,863
• Time in Patent Office: 523 Days
• Field of Search: 204/299 R, 252/315.1, 524/505, 524/520, 524/504, 524/507
• US Class Current: 204/455
• CPC Class Codes: G01N 27/44747 Composition of gel or of carrier mixture

Abstract

A viscous electrophoresis separation medium is disclosed. The medium is formed by a matrix formed by a copolymer composed of hydrophilic polymer segments having selected, substantially uniform segment lengths, and a plurality of hydrophobic polymer segments carried on, and spaced from one another at regular repeating intervals, by the hydrophilic polymer segments. Also disclosed is an electrophoresis method which employs the separation medium, and novel copolymers used in forming the medium.

Claims

1. An electrophoresis separation medium comprising

a matrix of aggregated, regular-repeating copolymers in an aqueous medium, said copolymers being composed of hydrophilic polymer segments having selected, substantially uniform segment lengths, and a plurality of hydrophobic polymer segments carried on and spaced from one another, at regular repeating intervals, by the hydrophilic polymer segments,

said medium being characterized by:

(i) the ability of the medium to effect a high-resolution electrophoretic separation of biopolymer molecules in a defined molecular size range, and

(ii) a concentration of the copolymer which is above the interpolymeric-aggregation transition concentration, defined by the concentration of copolymer sufficient to produce a marked rise in viscosity of an aqueous dispersion of the copolymer.

2. The medium of claim 1, wherein the copolymers have a structure selected from the group consisting of:

(a) a comb copolymer structure in which the hydrophobic polymer segments extend outwardly from a backbone composed of linked hydrophilic polymer segments;

(b) a block copolymer structure having an alternating linear sequence of the hydrophilic segments and hydrophobic segments; and

(c) a star copolymer structure in which the hydrophobic polymer segments are carried at the distal ends of the hydrophilic polymer segments which are attached at their proximal ends to a common anchor.

3. The medium of claim 1, wherein the hydrophilic polymer segments are selected from the group consisting of polyethylene oxide, polyglycolic acid, polylactic acid, polysaccharide, polyurethane, polyamades, polysulfonamide, and polysulfoxide.

4. The medium of claim 3, wherein the hydrophilic polymer segments are joined by linking moieties, and the hydrophobic polymer segments are attached to the linking moieties.

5. The medium of claim 1, wherein the hydrophobic polymer segments are fluorinated hydrocarbon chains.

6. The medium of claim 1, wherein the hydrophilic polymer segments are polyethylene oxide segments.

7. The medium of claim 6, wherein the copolymer is a comb copolymer structure in which fluorinated hydrocarbon segments extend outwardly from a backbone composed of linked polyethylene oxide segments.

8. The medium of claim 1, for use in separating DNA fragments within a selected size range, wherein the copolymers have a comb structure having a backbone composed of linked hydrophilic segments and hydrophobic segments spaced at substantially equal intervals along the backbone, at a selected distance between about 50 and 1,000 backbone chain atoms, for fractionation of DNA fragments in selected size ranges (i) of less than about 100 basepairs, at a selected backbone spacing of about 100 chain atoms, (ii) of greater than about 1,000 basepairs, at a selected backbone spacing between about 500-1,000 chain atoms, or (iii) of sizes between about 100-1,000 basepairs at a selected backbone spacing between about 100-800 chain atoms.

9. The method of claim 8, wherein the copolymer is a comb copolymer in which fluorinated hydrocarbon segments extend outwardly from a backbone composed of linked polyethylene oxide segments.

10. An electrophoresis system for separating biopolymers, comprising

a support defining an elongate channel connectable at opposite ends to opposing polarity terminals of a voltage source, and

contained within the channel, an electrophoresis separation medium comprising a matrix of aggregated regular alternating copolymers in an aqueous medium, said copolymers being composed of hydrophilic polymer segments having selected, substantially uniform segment lengths, and a plurality of hydrophobic polymer segments carried on and spaced from one another, at regular repeating intervals, by the hydrophilic polymer segments,

said medium being characterized by:

(i) the ability of the medium to effect a high-resolution electrophoretic separation of biopolymer molecules in a defined molecular size range, and

(ii) a concentration of the copolymer which is above the interpolymeric-aggregation transition concentration, defined by the concentration of copolymer sufficient to produce a marked rise in viscosity of an aqueous dispersion of the copolymer.

11. The system of claim 10, wherein the support is a capillary tube.

12. The system of claim 10, wherein the copolymers have a structure selected from the group consisting of:

(a) a comb copolymer structure in which the hydrophobic polymer segments extend outwardly from a backbone composed of linked hydrophilic polymer segments;

(b) a block copolymer structure having an alternating linear sequence of the hydrophilic segments and hydrophobic segments; and

(c) a star copolymer structure in which the hydrophobic polymer segments are carried at the distal ends of the hydrophilic polymer segments which are attached at their proximal ends to a common anchor.

13. The system of claim 10, wherein the hydrophilic polymer segments are selected from the group consisting of polyethylene oxide, polyglycolic acid, polylactic acid, polysaccharide, polyurethane, polyamides, polysulfonamide, and polysulfoxide.

14. The system of claim 13, wherein the hydrophobic polymer segments are fluorinated hydrocarbon chains.

15. The medium of claim 14, wherein the hydrophilic polymer segments are polyethylene oxide segments.