Methods and apparatus for rapid crystallization of biomolecules
First Claim
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1. A method for rapid crystallization of biomolecules, comprising:
- (a) providing at least one biomolecules species;
(b) providing at least one crystallization reactor comprising an isoelectric focusing buffer having a pH range, the pH range encompassing the pI of the at least one biomolecule species;
(c) bringing said at least one biomolecule species into contact with the at least one crystallization reactor;
(d) introducing an electric field at said at least one crystallization reactor thereby generating a concentrated solution of said at least one biomolecule species; and
(e) obtaining at least one crystal within said at least one crystallization reactor;
wherein the electric field is within the range of 50-2,000 V/cm.
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Abstract
The present invention relates to methods and apparatus for promoting rapid formation of biomolecule crystals from a solution of biomolecules, preferably proteins, wherein the protein solution undergoes rapid concentration according to its isoelectric point in an electric field. Protein crystallization according to the methods of the present invention takes place within a period of hours or less.
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Citations
55 Claims
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1. A method for rapid crystallization of biomolecules, comprising:
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(a) providing at least one biomolecules species; (b) providing at least one crystallization reactor comprising an isoelectric focusing buffer having a pH range, the pH range encompassing the pI of the at least one biomolecule species; (c) bringing said at least one biomolecule species into contact with the at least one crystallization reactor; (d) introducing an electric field at said at least one crystallization reactor thereby generating a concentrated solution of said at least one biomolecule species; and (e) obtaining at least one crystal within said at least one crystallization reactor; wherein the electric field is within the range of 50-2,000 V/cm. - View Dependent Claims (2, 3, 4, 5, 6, 7, 9, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20)
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8. A method for rapid crystallization of biomolecules, comprising:
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(a) providing at least one biomolecules species; b) providing at least one crystallization reactor comprising an isoelectric focusing buffer having a pH range, the pH range encompassing the pI of the at least one biomolecule species; (c) bringing said at least one biomolecule species into contact with the at least one crystallization reactor; (d) introducing an electric field at said at least one crystallization reactor thereby generating a concentrated solution of said at least one biomolecule species; and (e) obtaining at least one crystal within said at least one crystallization reactor; wherein the at least one biomolecule species in step (a) is immobilized onto a substrate comprising a porous web material.
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10. A method for rapid crystallization of biomolecules, comprising:
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(a) providing at least one biomolecules species; (b) providing at least one crystallization reactor comprising an isoelectric focusing buffer having a pH range, the pH range encompassing the pI of the at least one biomolecule species; (c) bringing said at least one biomolecule species into contact with the at least one crystallization reactor; (d) introducing an electric field at said at least one crystallization reactor thereby generating a concentrated solution of said at least one biomolecule species; and (e) obtaining at least one crystal within said at least one crystallization reactor; wherein the at least one crystallization reaction is linked, joined, or substantially contiguous to a solid substrate comprising a porous web material.
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21. A method for sorting a solution comprising a plurality of biomolecules and rapidly crystallizing at least one biomolecule species, comprising:
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(a) providing a medium comprising a plurality of biomolecules; (b) sorting the plurality of biomolecules on a substrate, thereby obtaining at least one locus on the substrate comprising at least one biomolecule species; (c) recovering a portion from said substrate, the portion comprising the at least one locus; (d) providing at least one crystallization reactor comprising an isoelectric focusing buffer having a pH range, the pH range encompassing the pI of the at least biomolecule; (e) bringing the portion of (c) into contact with the at least one crystallization reactor; (f) introducing an electric field at the at least one crystallization reactor thereby generating within said at least one crystallization reactor a concentrated solution of said at least one biomolecule species; and (g) obtaining at least one crystal within said at least one crystallization reactor of (f). - View Dependent Claims (22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34)
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35. An apparatus suitable for inducing rapid formation of biomolecule crystals, comprising:
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(a) a buffer chamber having an upper side and a lower side, the lower side being sealed with a bottom such that the buffer chamber encloses at least one buffer compartment capable of holding fluids; (b) at least one crystallization reactor contained within said buffer chamber, the at least one crystallization reactor comprising an isoelectric focusing buffer; (c) a device for generating an electrical field; (d) two salt bridges having two ends, one end of each salt bridge being in contact with one end of the holder and one end of each salt bridge being contained within the at least one buffer chamber; and
, optionally,(e) means for circulating fluids contained within the at least one buffer compartment. - View Dependent Claims (36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55)
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Specification