Process of high purity albumin production
AssigneeDelta Biotechnology Limited
InventorsAndrew Robert Goodey, Darrell Sleep, Hendrik Van Urk, Stephen Berezenko, John Rodney Woodrow, Richard Alan Johnson, Patricia Carol Wood, Stephen James Burton, Alan Victor Quirk
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Abstract
The present invention relates to a process for purifying the protein human serum albumin extracted from serum or recombinant human albumin produced by transforming a microorganism with a nucleotide coding sequence encoding the amino acid sequence of human serum albumin. The present invention provides highly purified albumin.
Claims67
A process for purifying albumin, the process comprising the steps of:
- (a) passing an impure albumin solution through a chromatographic material for which the albumin has no specific affinity under conditions such that the albumin will bind to the chromatographic material and low molecular weight species will pass directly through the material;
- and
- (b) eluting the bound albumin from the chromatographic material by passing through the chromatographic material a solution of a compound having a specific affinity for albumin, to yield a purified albumin solution.
A process for purifying albumin, comprising the steps of:
- (a) passing an albumin solution through a cation exchange matrix under conditions such that the albumin will bind to the matrix;
- (b) eluting from said matrix an albumin-containing cation exchange eluate;
- (c) passing said eluate through an affinity matrix comprising an albumin-binding compound;
- (d) eluting from the affinity matrix an albumin-containing affinity matrix eluate;
- (e) passing said eluate through a gel permeation matrix to obtain a fraction enriched in albumin;
- (f) passing said albumin-enriched fraction through an anion exchange matrix under conditions such that albumin will bind to the anion exchange matrix;
- and
- (g) eluting from said anion exchange matrix a purified albumin-containing product.
A process for purifying albumin, comprising the steps of:
- (a) passing an albumin solution through a cation exchange matrix under conditions such that the albumin will bind to the matrix;
- (b) eluting from the matrix an albumin-containing cation exchange eluate;
- (c) passing the cation exchange eluate through an anion exchange matrix under conditions such that the albumin will bind to the matrix;
- (d) eluting from the anion exchange matrix an albumin-containing anion exchange eluate;
- (e) passing the anion exchange eluate through an affinity matrix comprising an albumin-binding compound;
- (f) eluting from the affinity matrix an albumin-containing affinity matrix eluate;
- and
- (g) passing the affinity matrix eluate through a gel permeation matrix to obtain a fraction enriched in albumin.
A method of removing ammonium ions bound to albumin in an albumin solution, the method comprising (a) exposing the albumin solution to a solution of a counter-ion such that ammonium ions are displaced from the albumin and (b) removing the ammonium ions from the albumin solution.
A method of purifying intact albumin from a solution which contains the intact albumin and an approximately 45 kD albumin fragment, the method comprising (a) passing the solution through a chromatographic material comprising an immobilized dye having specific affinity for albumin such that the albumin in the solution binds to the dye;
- and (b) selectively eluting the bound intact albumin from the bound 45 kD albumin fragment, to yield purified intact albumin.
A process for purifying albumin, the process comprising the steps of:
- (a) subjecting an albumin solution to cation exchange chromatography in which the albumin is bound to a cation exchange material and then eluted therefrom;
- (b) treating the albumin eluted in step (a) by one or more processes selected from the group consisting of affinity chromatography, ultrafiltration, and gel filtration;
- and
- (c) treating the albumin obtained from step (b) to anion exchange chromatography in which the albumin is bound to an anion exchange material.
A process for purifying albumin, the process comprising the steps of:
- (a) subjecting an albumin solution to cation exchange chromatography in which the albumin is bound to a cation exchange material and then eluted therefrom;
- and
- (b) subjecting the albumin eluted from the cation exchange material in step (a) to anion exchange chromatography in which the albumin is bound to an anion exchange material without any intervening treatment between steps (a) and (b) or without any intervening treatment other than dilution of the albumin.
A process for purifying albumin in an impure albumin solution from a macromolecule with a carbohydrate moiety, the process comprising the steps of:
- (a) passing the impure albumin solution through a chromatographic material comprising a boronic acid or salt thereof such that the macromolecule with the carbohydrate moiety binds to the chromatographic material;
- and (b) eluting the albumin from the chromatographic material, to yield a purified albumin solution.
Dependencies are inferred from the claim text.