METHODS OF PURIFYING POLYPEPTIDES
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Abstract
The present invention relates to improved processes for purifying polypeptides of interest by increasing the amount of a polypeptide of interest bound to an ion-exchange matrix relative to the amount of one or more impurities bound to the ion-exchange matrix. This effect is achieved by adding a chemical compound in the process which by also binding to the ion-exchange matrix due to a change that is opposite to the change of the ion-exchange matrix, reduces the binding of impurities more than the binding of the polypeptide of interest.
1 Citation
60 Claims
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1-29. -29. (canceled)
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30. A process for the purification of a polypeptide of interest comprising ion-exchange chromatography, comprising the steps of
adding a chemical compound with the ability to complex metal ions to one or more of a) an equilibration fluid of the ion-exchange matrix, b) a loading fluid, which is applied to the ion-exchange matrix and which comprises the polypeptide of interest, or c) one or more washing fluids with a pH of about 3 to about 10, which is used to wash the ion-exchange matrix once the polypeptide of interest has bound to the ion-exchange matrix due to a charge that is opposite to the charge of the ion-exchange matrix; - and
eluting the polypeptide of interest using an elution fluid with a pH of from about 3 to about 10, thereby providing an increased ratio of the polypeptide of interest to one or more impurities in the eluate as compared to the same process where the chemical compound is not added, wherein the pH of the elution fluid and the pH of at least one washing fluid are different; the polypeptide of interest is a Vitamin K-dependent polypeptide; and the ion-exchange matrix is an anion-exchange matrix. - View Dependent Claims (31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60)
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Specification