Method for albumin purification
View Patent ↗The present invention is a method of purifying recombinant human serum albumin (rHSA) from a solution, which method comprises to subject a cell culture supernatant (CCS) comprising rHSA cation exchange on a bimodal high salt tolerant matrix; hydrophobic interaction chromatography (HIC); anion exchange; and recovering the purified rHSA. The bimodal high salt tolerant cation exchange matrix used enables performing a purification of a cell culture supernatant directly, in the sense that no further dilution thereof is necessary. Due to its high binding capacity, said bimodal cation exchange matrix also allows use of a smaller amount of matrix as compared to a corresponding conventional cation exchanger matrix. Accordingly, the present invention allows substantial savings as regards volumes and consequently operation costs.
1. A method of purifying recombinant human serum albumin (rHSA) from a cell culture supernatant (CCS), which method comprises:
(a) subjecting said CCS to cation exchange on a bimodal high salt tolerant matrix;
(b) followed by hydrophobic interaction chromatography (HIC);
(c) then anion exchange; and
(d) recovering the purified rHSA.
2. The method of claim 1 , wherein the conductivity of the CCS is above about 10 when applied to step (a).
3. The method of claim 1 , wherein the bimodal cation exchange matrix is capable of interacting with rHSA by charge interaction and by hydrogen bonding and/or hydrophobic interaction.
4. The method of claim 1 , further comprising heat treatment of the CCS before step (a).
5. The method of claim 1 , further comprising heat treatment of the product from step (a) before step (b) in the presence of a reducing agent.
6. The method of claim 1 , wherein step (b) utilizes a HIC matrix including phenyl, aliphatic and/or heterocyclic ligands.
7. The method of claim 1 , wherein the amount of cation exchange matrix used in step (a) is about half the amount of HIC matrix used in step (b).
8. The method of claim 1 , wherein step (c) utilizes a weak anion exchanger.
9. The method of claim 8 , wherein the ligand density of the weak anion exchanger is >50 μmol/ml gel/matrix.
10. The method of claim 9 , wherein the purified rHSA is recovered only from the bound fraction from step (c).
11. The method of claim 8 , wherein the ligand density of the weak anion exchanger is >100 μmol/ml gel/matrix.
12. The method of claim 8 , wherein the ligand density of the weak anion exchanger is about 160 μmol/ml gel/matrix.