Selective removal of a protein from a mixture of proteins using activated carbon by adjusting solution conditions
The present invention provides novel and improved protein purification processes which incorporate certain types of carbonaceous materials and result in effective and selective removal of certain undesirable impurities without adversely effecting the yield of the desired protein product.
1. A method of selectively removing a proteinaceous impurity from a sample comprising at least an immunoglobulin and the proteinaceous impurity, the method comprising the steps of:
(a) providing a sample comprising at least an immunoglobulin and a proteinaceous impurity;
(b) adjusting the solution pH of the sample, such that the pH is within 2.0 pH units of the isoelectric point of the proteinaceous impurity to be selectively removed;
(c) contacting the sample with activated carbon, wherein the activated carbon selectively binds the proteinaceous impurity to be selectively removed; and
(d) removing the activated carbon from the sample,
thereby resulting in selective-removal of the activated carbon bound proteinaceous impurity from the sample and an increase in the concentration of the immunoglobulin to the proteinaceous impurity in the sample of 70% or more when measured.
2. The method of claim 1 , wherein the pH is within 1.0 pH unit of the isoelectric point of the proteinaceous impurity to be selectively removed.
3. The method of claim 1 , wherein the immunoglobulin is a monoclonal antibody.
4. The method of claim 1 , wherein the immunoglobulin is a polyclonal antibody.
5. The method of claim 1 , wherein the removing step comprises filtration or centrifugation.
6. The method of claim 1 , wherein the immunoglobulin is a recombinant immunoglobulin.
7. The method of claim 1 , wherein the sample comprises a cell culture feed.
8. The method of claim 7 , wherein the cell culture feed is a CHO cell culture feed.
9. The method of claim 1 , wherein the immunoglobulin is expressed in a mammalian expression system.
10. The method of claim 1 , wherein the immunoglobulin is expressed in a non-mammalian expression system.
11. The method of claim 1 , wherein the sample is subjected to a clarification step prior to the adjusting step.
12. The method of claim 11 , wherein the clarification step is selected from the group consisting of centrifugation, settling, depth or screen filtration, complexing with flocculants, and pH change.
13. The method of claim 1 , wherein the increase in the concentration of the immunoglobulin to the proteinaceous impurity in the sample is 80% or more.
14. The method of claim 1 , wherein the increase in the concentration of the immunoglobulin to the proteinaceous impurity in the sample is 90% or more.
15. The method of claim 1 , the method further comprising a sample with multiple impurities and, between steps a) and b), determining the isoelectric focusing point or points of the multiple impurities in the sample and identifying which proteinaceous impurity or impurities are to be selectively removed from the sample.
16. The method of claim 15 , wherein said isoelectric point or points is determined by focusing electrophoresis gel, capillary isoelectric focusing or two-dimensional gel electrophoresis.