Separation media for electrophoresis
First Claim
1. In an electrophoretic device comprising an electrophoretic medium, the improvement which comprises:
- an electrophoretic medium comprising a dispersion of particulate chemically-crosslinked microgels having particle sizes with a distribution of particles no smaller than about 10 nm and no larger than about 2 mm, wherein the weight concentration of said microgels in said dispersion is between about 1% and 50% and wherein said medium is reversibly convertible between a pumpable low viscosity state and an electrophoretically sieving high viscosity state, said high viscosity state characterized by a viscosity of at least 100% greater than the viscosity of said medium in said low viscosity state, and wherein the conversion of said low viscosity state to said high viscosity state can be affected by a change in temperature, pH, solvent concentration, ionic strength, presence and concentration of a specific ion, or other property or combinations of properties of said medium.
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Accused Products
Abstract
Separation media for electrophoresis, and methods of filling and flushing of electrophoretic devices such as capillaries are described. By preparing submicron to above-micron sized cross-linked gel particles and using gel swelling equilibrium concepts, such devices can be easily filled and flushed. Gel particles can be prepared by inverse suspension, precipitation and suspension polymerization. These particles can be swollen and collapsed by small changes in temperature, pH, and ionic strength of solvent. Other approaches involve the formation of reversible cross-links by use of polyelectrolyte complexes, chelating agents or copolymers of hydrophobic and hydrophilic repeat units. Finally, reversibly solubilized systems may be used to change the viscosity of the media.
185 Citations
24 Claims
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1. In an electrophoretic device comprising an electrophoretic medium, the improvement which comprises:
an electrophoretic medium comprising a dispersion of particulate chemically-crosslinked microgels having particle sizes with a distribution of particles no smaller than about 10 nm and no larger than about 2 mm, wherein the weight concentration of said microgels in said dispersion is between about 1% and 50% and wherein said medium is reversibly convertible between a pumpable low viscosity state and an electrophoretically sieving high viscosity state, said high viscosity state characterized by a viscosity of at least 100% greater than the viscosity of said medium in said low viscosity state, and wherein the conversion of said low viscosity state to said high viscosity state can be affected by a change in temperature, pH, solvent concentration, ionic strength, presence and concentration of a specific ion, or other property or combinations of properties of said medium. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 13, 14, 15, 16, 17, 18, 19, 20)
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10. In an electrophoretic device comprising an electrophoretic medium, the improvement which comprises:
an electrophoretic medium comprising polymer chains having hydrophobic and hydrophilic moieties, wherein said medium is reversibly convertible between an electrophoretically sieving high viscosity state and a pumpable low viscosity state, said high viscosity state characterized by a viscosity of at least 100% greater than the viscosity of said medium in said low viscosity state, and wherein the conversion of said low viscosity state to said high viscosity state can be affected by a change in temperature, pH, solvent concentration, ionic strength, presence and concentration of a specific ion, or other property or combinations of properties of said medium which cause coalescence of said hydrophobic or hydrophilic groups on contiguous polymeric chains to form effective crosslinks of chains in said medium.
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11. In an electrophoretic device comprising an electrophoretic medium, the improvement which comprises:
an electrophoretic medium comprising polymer chains having ionic moieties and counter-ions, wherein said medium is reversibly convertible between an electrophoretically sieving high viscosity state and a pumpable low viscosity state, said high viscosity state characterized as having a viscosity of at least 100% greater as compared to the viscosity of said medium in said low viscosity state, and wherein the conversion of said low viscosity state to said high viscosity state is affected by exchange of said counter-ions with counter-ions of higher valency whereby ionic moieties on contiguous polymeric chains associate with said counter-ions of higher valency to form effective crosslinks of chains in said medium.
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12. In an electrophoretic device comprising an electrophoretic medium, the improvement which comprises:
an electrophoretic medium comprising polymer chains having chelatable ligand, wherein said medium is reversibly convertible between an electrophoretically sieving high viscosity state and a pumpable low viscosity state, said high viscosity state characterized as having a viscosity of at least 100% greater as compared to the viscosity of said medium in said low viscosity state, and wherein the conversion of said low viscosity state to said high viscosity state is affected by addition of a chelating agent whereby chelatable ligands on contiguous polymeric chains associate with said chelating agent to form effective crosslinks of chains in said medium.
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21. In an electrophoretic device comprising an electrophoretic medium, the improvement which comprises:
said electrophoretic medium comprising a composition reversibly convertible going from a first pumpable state to a second continuous electrophoretic gel state, wherein in said first pumpable state said composition is discrete particles of a cross-linked N-substituted polyacrylamide of a size in the range of 10 nm to 2 mm in an aqueous medium; and
in said second state said particles are swollen and form a continuous gel, wherein said change from one state to another state occurs with a change in temperature.- View Dependent Claims (22, 23)
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24. In an electrophoretic device comprising an electrophoretic medium, the improvement which comprises:
an electrophoretic medium comprising a dispersion of a particulate chemically-crosslinked microgel having particle sizes with a distribution of particles no smaller than about 10 nm and no larger than about 2 mm, wherein the weight concentration of said microgel in said dispersion is between about 1% and 50%, agarose, and wherein said medium is reversibly convertible between a pumpable low viscosity state and an electrophoretically sieving gel slab state, and wherein the conversion of said low viscosity state to said gel slab state occurs with a change in temperature.
Specification