Process for the purification of serum albumin
AssigneeGist-Brocades N.V.
InventorsCornelis Jacobus van der Laken, Marcellinus Petrus Johannes Piet
Life of this patent
Estimating the term…
Abstract
Recombinantly produced serum albumin is purified in a series of steps, optionally by incubation with an anion-exchange adsorbent, followed by affinity chromatography employing a hydrophobic solid phase and using a water-soluble lipid anion as desorbens in the aqueous phase. This immobile phase comprises a carrier coupled to a 2-mercapto or 2-hydroxy alkanoic acid.
Claims15
A method for producing purified recombinant serum albumin substantially free of albumin degradation products, said method comprising the steps of:
- (a) incubating at about physiologic pH a product medium comprising a clarified fermentation broth or clarified cell lysate comprising recombinant serum albumin with an anion-exchange agent;
- (b) eluting said anion-exchange agent with a buffer solution having an acid pH and collecting first eluate fractions which comprise said recombinant serum albumin;
- (c) contacting at about physiologic pH said first eluate fractions with a chromatography system comprising a lipophilic immobile phase and an aqueous mobile phase, wherein said immobile phase comprises a carrier activated by an epoxide compound coupled to a 2-mercapto C4-C14 alkanoic compound or 2-hydroxy C4-C14 alkanoic compound selected from the group consisting of an acid, a salt and an ester of either of said compounds;
- (d) eluting recombinant serum albumin from said chromatography system by adding as desorbens a water soluble lipid anion to said aqueous mobile phase and collecting second eluate fractions comprising recombinant serum albumin;
- and
- (e) isolating from said second eluate fractions purified recombinant serum albumin substantially free of albumin degradation products.
A method for producing purified recombinant serum albumin, said method comprising the steps:
- (a) incubating at about physiologic pH a product medium comprising a clarified fermentation broth or clarified cell lysate comprising recombinant serum albumin with an anion-exchange agent;
- (b) eluting said anion-exchange agent with a buffer solution having an acid pH and collecting first eluate fractions which comprise said recombinant serum albumin;
- (c) contacting at about physiologic pH said first eluate fractions with a chromatography system comprising a lipophilic immobile phase and an aqueous mobile phase, wherein said immobile phase comprises a carrier activated by an epoxide compound coupled to a 2-mercapto C4-C14 alkanoic compound or 2-hydroxy C4-C14 alkanoic compound selected from the group consisting of an acid, a salt and an ester of either of said compounds;
- (d) eluting recombinant serum albumin from said chromatography system by adding as desorbens a water soluble lipid anion to said aqueous mobile phase and collecting second eluate fractions comprising recombinant serum albumin;
- and
- (e) isolating from said second eluate fractions purified recombinant serum albumin substantially free of cellular components of microorganisms or cell lines.
A method for purifying recombinant serum albumin substantially free of albumin degradation products, said method comprising:
- isolating purified recombinant serum albumin from an affinity chromatography system, wherein said affinity chromatography system comprises a lipophilic immobile phase and an aqueous mobile phase, wherein said lipophilic immobile phase comprises a carrier activated by an epoxide compound coupled to a 2-mercapto C4-C14 alkanoic compound or 2-hydroxy C4-C14 alkanoic compound selected from the group consisting of an acid, a salt and an ester of either of said compounds.
Dependencies are inferred from the claim text.