Protein purification
AssigneeGenencor International, Inc.
InventorsRobin Fong, Meng Heng
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Abstract
Described herein are methods for purifying a protein of interest from a mixture of proteins wherein the mixture comprises the protein of interest and a fusion analog thereof. The method begins with a recovery in which filtration is used to remove cells and concentrate the protein of interest. Liquid chromatography is then used to remove fusion analogs and other contaminants resulting in a composition substantially free of contaminants. Substantially pure compositions of the protein of interest find use in therapeutic preparations.
Claims25
A method of purifying a protein of interest from its fusion analog, said method comprising:
- a. Obtaining a protein solution comprising the protein of interest and its fusion analog;
- b. Adjusting the pH and/or ionic strength of the protein solution with an appropriate buffer for the HClC resin used in step c;
- c. Contacting the protein solution with an HClC resin column for a time sufficient to allow binding of the protein of interest and its fusion analog to the resin;
- d. Washing the HClC resin with an appropriate buffer;
- e. Eluting the protein of interest from the HClC resin by a pH gradient;
- wherein said protein of interest is substantially free of its fusion analog.
A method of purifying an immunoglobulin, said method comprising:
- a. Obtaining a protein solution comprising the immunoglobulin;
- b. Adjusting the pH and/or ionic strength of the protein solution with an appropriate buffer for the HClC resin used in step c;
- c. Contacting the protein solution with an HClC resin for a time sufficient to allow binding of the immunoglobulin to the resin;
- d. Washing the HClC resin of an appropriate buffer;
- e. Eluting the immunoglobulin from the HClC resin by a pH gradient.
Dependencies are inferred from the claim text.