Detection of cho-mif contaminations

Abstract

The invention concerns a detection method to enable the detection of complexes formed between antibodies and antigen which is endogenous to the production cell line, e.g. CHO MIF in a final product.

1 .- 13 . (canceled) 
     
     
         14 . A method for the detection of Chinese hamster ovary (CHO)-migration inhibitory factor (MIF) contaminations in a monoclonal anti-(h)MIF antibody preparation, comprising contacting the anti-(h)MIF antibody preparation with a monoclonal rabbit anti-CHO-MIF antibody, and detecting CHO-MIF contaminations. 
     
     
         15 . The method according to  claim 14  wherein the CHO-MIF contaminates a final CHO cell produced monoclonal anti-(h)MIF antibody-preparation or a preparation of antigen-binding portions thereof. 
     
     
         16 . The method according to  claim 14  wherein the CHO-MIF is endogenous CHO-MIF produced by CHO cells, per se or CHO MIF complexed with the anti-(h)MIF antibody. 
     
     
         17 . The method according to  claim 14  wherein the detection step is carried out by a semi-quantitative Western Blot analysis, or by an ELISA. 
     
     
         18 . The method according to  claim 17  wherein the ELISA is preferably a quantitative ELISA. 
     
     
         19 . The method of any one of  claim 14  wherein said monoclonal antibody is A5- antibody, which comprises a heavy chain characterized by SEQ ID NO:18, and a light chain characterized by SEQ ID NO: 20. 
     
     
         20 . Use of a monoclonal rabbit anti-CHO-MIF antibody, preferably the A5-antibody, for the detection of CHO-MIF contaminations during production of monoclonal anti-(h)MIF antibodies or antigen-binding fragments thereof or in the final preparation of monoclonal anti-(h)MIF antibody or antigen-binding portions thereof. 
     
     
         21 . The use according to  claim 20  wherein the detection step is carried out as a semi-quantitative Western Blot analysis or as an ELISA. 
     
     
         22 . The use according to  claim 21  wherein the ELISA is preferably a quantitative ELISA. 
     
     
         23 . The use according to  claim 20  wherein the anti-(h)MIF antibody is selected from the group consisting of RAB4, RAB0, RAB9, RAM4, RAM0 and RAM9, wherein:
 RAB4 comprises a light chain characterized by SEQ ID NO: 2, and a heavy chain characterized by SEQ ID NO: 6; 
 RAM4 comprises a light chain characterized by SEQ ID NO: 14, and a heavy chain characterized by SEQ ID NO: 13; 
 RAB9 comprises a light chain characterized by SEQ ID NO: 1, and a heavy chain characterized by SEQ ID NO: 5; 
 RAM9 comprises a light chain characterized by SEQ ID NO: 12, and a heavy chain characterized by SEQ ID NO: 11; 
 RAB0 comprises a light chain characterized by SEQ ID NO: 3, and a heavy chain characterized by SEQ ID NO: 7; and 
 RAM0 comprises a light chain characterized by SEQ ID NO: 10, and a heavy chain characterized by SEQ ID NO: 9. 
 
     
     
         24 . The use according to  claim 23  wherein the anti-(h)MIF antibody is RAM9, which comprises a light chain characterized by SEQ ID NO: 12, and a heavy chain characterized by SEQ ID NO: 11. 
     
     
         25 . Essentially Chinese hamster ovary (CHO)-migration inhibitory factor (MIF) free anti-(h)MIF antibody preparation as obtainable by the method of  claim 14 . 
     
     
         26 . Recombinant anti-(h)MIF antibody preparation, produced in a CHO cell line, characterized in that said preparation is essentially free of CHO-MIF, wherein said preparation is produced by a method which comprises, preferably as a quality control step, the method of detection of claim  1 . 
     
     
         27 . The anti-(h)MIF antibody preparation according to  claim 26  which is essentially free of CHO MIF, wherein the anti-(h)MIF antibody is selected from the group of RAB4, RAB0, RAB9, RAM4, RAM0 and/or RAM9 wherein:
 RAB4 comprises a light chain characterized by SEQ ID NO: 2, and a heavy chain characterized by SEQ ID NO: 6; 
 RAM4 comprises a light chain characterized by SEQ ID NO: 14, and a heavy chain characterized by SEQ ID NO: 13; 
 RAB9 comprises a light chain characterized by SEQ ID NO: 1, and a heavy chain characterized by SEQ ID NO: 5; 
 RAM9 comprises a light chain characterized by SEQ ID NO: 12, and a heavy chain characterized by SEQ ID NO: 11; 
 RAB0 comprises a light chain characterized by SEQ ID NO: 3, and a heavy chain characterized by SEQ ID NO: 7; and 
 RAM0 comprises a light chain characterized by SEQ ID NO: 10, and a heavy chain characterized by SEQ ID NO: 9. 
 
     
     
         28 . The anti-(h)MIF antibody preparation according to  claim 27 , wherein the anti-(h)MIF antibody is RAM9, which comprises a light chain characterized by SEQ ID NO:12, and a heavy chain characterized by SEQ ID NO: 11. 
     
     
         29 . A method for the production of an anti-(h)MIF antibody according to  claim 26 . 
     
     
         30 . The method for the production of an anti-(h)MIF antibody of  claim 29 , wherein the detection method is a quality control step for the verification that a CHO MIF content in the anti-(h)MIF antibody preparation is equal to or below 4 ppm.