METHOD FOR THE PURIFICATION OF ANTIBODIES

摘要

A method for the purification of immunoglobulins by ion exchange chromatography is described. The chromatographic method uses a weak ion exchange resin and a single step elution process for the purification of an immunoglobulin. Additionally a method for the determination of the salt concentration for the single step elution of an immunoglobulin from an ion exchange resin is described.

主权项

1. A method for purifying a monomeric monoclonal antibody from aggregates thereof, wherein the method comprises:
a) providing a solution comprising a monoclonal antibody and a buffer substance; b) bringing the solution and a weak cation exchange material in contact under conditions whereby the monoclonal antibody binds to the weak cation exchange material; and c) recovering the monomeric monoclonal antibody from the weak cation exchange material in a single step by using a solution comprising a buffer substance and a salt wherein the conductivity of said solution is increased by changing one condition all at once from a starting value to a final value so as to obtain said monoclonal antibody purified from aggregates thereof; and wherein the method comprises purification of the monoclonal antibody by a protein A affinity chromatography prior to step a); and wherein the weak cation exchange material is a carboxy-methyl weak cation exchange material; wherein the monoclonal antibody is a member of the immunoglobulin class G; wherein the solution in the recovery step c) has a pH value of from pH 3.0 to pH 7.0; wherein the salt in step c) is selected from the group consisting of sodium chloride, sodium sulphate, potassium chloride, potassium sulfate, salts of citric acid, salts of phosphoric acid, and mixtures of these components; and wherein the buffer substance has a concentration range between 5 mM and 100 mM.