Antibody formulations and methods

Abstract

The invention provides antibody formulations and methods useful for prophylaxis or treatment of synucleinopathies, including Parkinson's disease.

1 - 48 . (canceled) 
     
     
         49 . A method of purifying a humanized, veneered, or chimeric 9E4 antibody expressed from CHO cells, comprising:
 (a) performing protein-A solid phase chromatography, wherein the 9E4 antibody preferentially binds the solid phase relative to impurities;   (b) performing anion-exchange solid phase chromatography, wherein the 9E4 antibody preferentially elutes from the solid phase relative to impurities;   (c) performing cation-exchange solid phase chromatograph, wherein the 9E4 antibody preferentially binds to the solid phase relative to impurities;   (d) performing a viral inactivation and/or removal step;   (e) performing a filtration step; and   (f) performing a concentration and resuspension step.   
     
     
         50 . The method of  claim 49 , wherein the protein-A, anion-exchange and cation exchange steps are performed in that order. 
     
     
         51 . The method of  claim 50 , wherein a viral inactivation step is performed between steps (a) and (b) and a viral removal step is performed after step (c). 
     
     
         52 . The method of  claim 51 , wherein a filtration step is performed between the viral inactivation step and the anion exchange step and after the cation exchange step. 
     
     
         53 . A method of purifying a humanized, veneered, or chimeric 9E4 antibody expressed from CHO cells, comprising:
 (a) loading culture medium from the cells expressing the 9E4 antibody on to a protein A column, wherein humanized 9E4 binds to the column, and reducing the pH to elute a fraction containing the 9E4 antibody;   (b) incubating the fraction from step (a) under acidic conditions to inactivate viruses;   (c) subjecting the fraction after step (b) to depth filtration to reduce particles;   (d) subjecting the filtrate obtained from step (c) containing the 9E4 antibody to anion exchange solid phase chromatography, wherein impurities in the fraction bind to the column and the 9E4 antibody elutes through the column;   (e) subjecting a fraction from step (d) containing the 9E4 antibody to cation exchange solid phase chromatography wherein the 9E4 antibody binds to the column and is eluted by raising the salt concentration;   (f) subjecting a fraction from step (e) containing the 9E4 antibody to viral filtration; and   (g) subjecting the filtrate containing 9E4 antibody from step (f) to ultra/diafiltration to concentrate and resuspend the 9E4 antibody.   
     
     
         54 . The method of  claim 53 , wherein the anion exchange column is Q-Sepharose fast flow resin. 
     
     
         55 . The method of  claim 53 , wherein in step (a) the pH is reduced from 6-8 to 2.5-3.5 
     
     
         56 . The method of  claim 53 , wherein step (b) is performed at pH3.5. 
     
     
         57 . The method of  claim 53 , wherein the filter in step (f) is a 0.1 micron filter. 
     
     
         58 - 64 . (canceled)