Claims
- 1. A process for protein purification in a high salt solution, comprising the steps of:
a. increasing the dynamic binding capacity of a resin by adding polyethylene glycol (PEG) to the high salt solution; and, d. contacting the resin with the high salt solution comprising at least one protein.
- 2. The process of claim 1 wherein the high salt solution is a cell culture broth.
- 3. The process of claim 1 wherein the PEG is chosen with a molecular weight between about PEG 400 and about PEG 10000, and mixtures thereof.
- 4. The process of claim 1 wherein the PEG is PEG 4600.
- 5. The process of claim 1 wherein the ion exchange resin is a cationic exchange resin.
- 6. The process of claim 5 wherein the conductivity of the solution is greater than about 9 mS/cm.
- 7. The process of claim 1 wherein the ion exchange resin is an anionic exchange resin.
- 8. The process of claim 7 wherein the conductivity of the solution is greater than about 11 mS/cm.
- 9. The process of claim 1 wherein the ion exchange occurs about a column.
- 10. The process of claim wherein the step of contacting is at a pH in a range of about 4.0 to about 10.0.
- 11. The process of claim 1 wherein the solution is diluted.
- 12. The process of claim 1 wherein the ion exchanger is a cation exchanger at a pH of about 5.0.
- 13. The process of claim 1 wherein the step of contacting the resin comprises contacting the resin with a solution comprising any one of bovine γ-globulin, bovine serum albumin, and lysozyme.
- 14. The process of claim 1 wherein the percentage PEG added is between about 0.5% W/v PEG and about 6% w/v PEG.
- 15. The process of claim 1 further comprising determining an ideal concentration of PEG for a particular protein by adding progressively greater concentrations of PEG to facilitate binding during loading of the resin until an acceptable amount of precipitation is observed.
- 16. The process of claim 1 further comprising filtering the load.
- 17. The process of claim 1 wherein the step of loading the resin further comprises equilibrating the resin with a binding buffer.
- 18. A process for the separation of a protein of interest from a cell culture broth added to an ion exchange column comprising the steps of:
a. increasing the dynamic binding capacity of an ion exchange resin by adding polyethylene glycol (PEG) to a high salt solution such as cell culture broth; b. contacting the resin with the cell culture broth whereby the protein of interest is at least partially captured; and, c. separating the protein of interest.
- 19. The process of claim 18 wherein the protein is a recombinant protein.
- 20. The process of claim 18 wherein the ion exchange column is either a cation exchange column or an anion exchange column.
- 21. The process of claim 18 further comprising the step of determining an ideal concentration of PEG for a particular protein by adding progressively greater concentrations of PEG to facilitate binding during loading of the resin until an acceptable amount of precipitation is noted.
- 22. A solution comprising a solution comprising a cell culture broth that has high salinity, a PEG which is between about 1% w/v and 8% w/v, and a protein; and a resin,
whereby said solution increases the dynamic binding capacity of the resin, thereby allowing the protein to be captured by the resin.
- 23. The solution of claim 22 wherein the resin is a cationic resin and the conductivity of the solution is greater than about 9.
- 24. The solution of claim 22 wherein the resin is an anionic resin and the conductivity of the solution is greater than about 11.
RELATED APPLICATION
[0001] This application is related to provisional application serial No. 06/401,570, filed on Aug. 6, 2002, titled Increased Dynamic Binding Capacity in Ion Exchange Chromatography by Addition of Polyethylene Glycol.
Provisional Applications (1)
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Number |
Date |
Country |
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60401570 |
Aug 2002 |
US |