Wash solution comprising arginine for the affinity chromatography step in antibody purification

Abstract

The invention provides a washing method for affinity chromatography in which a wash solution comprising arginine, or an arginine derivative, and a nonbuffering salt, preferably at high pH, greater than 8.0, is effective in removing impurities, such as high molecular weight species and host cell proteins, while also increasing product concentration in the eluate and maintaining a high percent yield of recovered product.

What is claimed is: 
     
         1 . A method of producing a purified antibody, antibody fragment, or Fc fusion protein, using an affinity chromatography (AC) matrix to which the antibody, antibody fragment, or Fc fusion protein is bound, the method comprising:
 (a) loading loading a mixture comprising the antibody, antibody fragment, or Fc fusion protein onto the AC matrix; and   (b) washing the AC matrix with one wash solution comprising arginine and a halogen salt selected from the group consisting of sodium chloride (NaCl), magnesium chloride (MgCl 2 ) and potassium chloride (KCl), prior to elution of the antibody, antibody fragment, or Fc fusion protein from the AC matrix.   
     
     
         2 . A method of producing a purified antibody, or antibody fragment, using a Protein A column, the method comprising
 (a) loading a mixture comprising the antibody, or antibody fragment, onto the Protein A column;   (b) washing the Protein A column with a wash solution comprising arginine at a concentration in a range of 0.05-0.85 M, and (ii) sodium chloride, at a concentration in a range of 0.1-2.0 M, wherein the wash solution removes impurities from the Protein A column; and   (c) eluting the antibody, or antibody fragment, from the Protein A column.   
     
     
         3 . The method of  claim 1 , wherein the AC matrix is a Protein A column. 
     
     
         4 . The method of  claim 1 , wherein the AC matrix is a Protein A column. 
     
     
         5 . The method of  claim 1 , wherein the wash solution removes impurities from the AC matrix. 
     
     
         6 . The method of  claim 5 , wherein the impurities comprise host cell proteins (HCPs). 
     
     
         7 . The method of  claim 1 , wherein the arginine is at a concentration of or about 0.25 M. 
     
     
         8 . The method of  claim 1 , wherein the arginine is arginine-HCl. 
     
     
         9 . The method of  claim 1 , wherein the halogen salt is at a concentration of or about 1 M. 
     
     
         10 . The method of  claim 9 , wherein the halogen salt is sodium chloride (NaCl). 
     
     
         11 . The method of  claim 1 , wherein the pH of the wash solution is greater than 8.0.