Nucleotide sugar purification using membranes
First Claim
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1. A method of removing essentially all of a contaminant from a mixture comprising the contaminant and a nucleotide sugar comprising a branched PEG structure, the method comprising:
- contacting the mixture with a first membrane for a length of time sufficient to allow essentially all of the contaminant to pass through the first membrane whereinthe mixture has a pH such that the first membrane and the nucleotide sugar comprising a branched PEG structure have a net charge of the same sign and the contaminant has a net charge which is neutral or a sign which is opposite of the sign of the net charge for the first membrane; and
the first membrane has a molecular weight cut-off that is less than the molecular weight of the nucleotide sugar comprising a branched PEG structure; and
wherein the branched PEG structure has a formula of;
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Abstract
The invention provides methods of removing contaminants from a mixture of a desired product and contaminants by pH adjustments and molecular weight cut-offs. The contaminants include phosphate groups, magnesium sulfate, sodium pyruvate and tetrasodium pyrophosphate groups. The desired product includes nucleotide sugars, glycolipids, LnNT, sialyl lactose, and salts.
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Citations
21 Claims
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1. A method of removing essentially all of a contaminant from a mixture comprising the contaminant and a nucleotide sugar comprising a branched PEG structure, the method comprising:
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contacting the mixture with a first membrane for a length of time sufficient to allow essentially all of the contaminant to pass through the first membrane wherein the mixture has a pH such that the first membrane and the nucleotide sugar comprising a branched PEG structure have a net charge of the same sign and the contaminant has a net charge which is neutral or a sign which is opposite of the sign of the net charge for the first membrane; and the first membrane has a molecular weight cut-off that is less than the molecular weight of the nucleotide sugar comprising a branched PEG structure; and wherein the branched PEG structure has a formula of; - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 17, 18, 19)
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13. A method of purifying a nucleotide sugar comprising a branched PEG structure from a mixture comprising a phosphorus-containing contaminant and the nucleotide sugar comprising a branched PEG structure, the method comprising:
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contacting the mixture with a first membrane for a length of time sufficient to allow essentially all of the phosphorus-containing contaminant to pass through the first membrane wherein the mixture has a pH such that the first membrane and the nucleotide sugar comprising a branched PEG structure have a net negative charge and the contaminant has a net charge which is a neutral or a positive charge; and the first membrane has a molecular weight cut-off that is less than the molecular weight of the nucleotide sugar comprising a branched PEG structure; and wherein the branched PEG structure has a formula of; - View Dependent Claims (14, 15, 20)
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16. A method of removing essentially all of a contaminant from a mixture comprising the contaminant and a nucleotide sugar comprising a branched PEG structure, the method comprising:
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contacting the mixture with a first membrane for a length of time sufficient to allow essentially all of the contaminant to pass through the first membrane wherein the mixture has a pH such that the first membrane and the nucleotide sugar comprising a branched PEG structure have a net charge of the same sign and the contaminant is at its isoelectric point; and the first membrane has a molecular weight cut-off that is less than the molecular weight of the nucleotide sugar comprising a branched PEG structure; and wherein the branched PEG structure has a formula of; - View Dependent Claims (21)
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Specification