Method for purifying granulocyte colony stimulating factor
AssigneeAmgen Inc.
InventorsThomas C. Boone, Allan L. Miller, Jeffrey W. Andresen
Life of this patent
Estimating the term…
Abstract
Processes for isolating and purifying granulocyte colony stimulating factor (G-CSF) from a G-CSF producing microorganism are disclosed. The simplified processes include steps of lysing the microorganism and separating insoluble material containing G-CSF from soluble proteinaceous material; extracting the material with deoxycholate (optionally); solubilizing and oxidizing the G-CSF in the presence of a denaturant solubilizing agent and an oxidizing agent; removing the denaturant solubilizing agent from the G-CSF; subjecting the G-CSF to ion exchange chromatography; and recovering the purified G-CSF; yet excludes other cumbersome purification steps.
Claims7
A process for isolating and purifying granulocyte colony stimulating factor (G-CSF) from a G-CSF producing microorganism consisting of:
- 1) lysing the microorganism and separating insoluble material containing G-CSF from soluble proteinaceous material;
- 2) solubilizing and oxidizing the G-CSF in the presence of a denaturant solubilizing agent and an oxidizing agent;
- 3) removing the denaturant solubilizing agent from the G-CSF;
- 4) subjecting the G-CSF to ion exchange chromatography;
- and
- 5) recovering purified G-CSF.
A process for isolating and purifying granulocyte colony stimulating factor (G-CSF) from a G-CSF producing microorganism consisting of:
- 1) lysing the microorganism and separating insoluble material containing G-CSF from soluble proteinaceous material;
- 2) extracting the material with deoxycholate;
- 3) solubilizing and oxidizing the G-CSF in the presence of a denaturant solubilizing agent and an oxidizing agent;
- 4) removing the denaturant solubilizing agent from the G-SCF;
- 5) subjecting the G-CSF to ion exchange chromatography;
- and
- 6) recovering the purified G-CSF.
Dependencies are inferred from the claim text.