Characterization of cho-mif gene and protein, and use thereof

Abstract

The present invention is concerned with the specific and highly sensitive detection of specific CHO-MIF complexes in the production of anti-MIF antibodies. The present invention is further concerned with the provision of specific antibodies which can be used for a CHO-MIF detection method. The present invention is also concerned with a CHO MIF knock-out cell line and use thereof. The present invention also provides preparations obtained from recombinant production in CHO cell lines which are essentially free of CHO-MIF.

1 .- 7 . (canceled) 
     
     
         8 . A Chinese hamster ovary (CHO)-macrophage migration inhibitory factor (MIF) knock-out cell line, wherein the CHO-MIF gene is knocked out. 
     
     
         9 . A CHO-MIF knock-out cell line according to  claim 8 , wherein said CHO MIF knock-out cell line comprises one or more plasmids selected from the group consisting of: DSM 25110, DSM 25112, DSM 25111, DSM 25113, DSM 25114, DSM 25115, DSM 25859, DSM 25860, DSM 25861, DSM 25862, DSM 25863 and DSM 25864. 
     
     
         10 . The CHO-MIF knock-out CHO cell line according to  claim 8 , wherein the cell line comprises the plasmids DSM 25110 and DSM 25112 or DSM 25861 and DSM 25862. 
     
     
         11 . The CHO-MIF knock-out CHO cell line according to  claim 8 , wherein the cell line comprises the plasmids DSM 25111 and DSM 25113 or DSM 25859 and DSM 25860. 
     
     
         12 . The CHO-MIF knock-out CHO cell line according to  claim 8 , wherein the cell line comprises the plasmids DSM 25114 and DSM 25115 or DSM 25863 and DSM 25864. 
     
     
         13 . The use of the CHO-MIF knock-out CHO cell line according to  claim 8  for the production of a preparation of monoclonal anti-MIF antibodies or binding fragments thereof, preferably for the production of an antibody selected from the group consisting of:
 i. a RAB9 antibody, characterized by a light chain sequence as deposited by way of plasmid deposition with deposit number DSM 25111 and a heavy chain sequence as deposited by way of plasmid deposition with deposit number DSM 25113, 
 ii. a RAB4 antibody, characterized by a light chain sequence as deposited by way of plasmid deposition with deposit number DSM 25110 and a heavy chain sequence as deposited by way of plasmid deposition with deposit number DSM 25112, 
 iii. a RAB0 antibody, characterized by a light chain sequence as deposited by way of plasmid deposition with deposit number DSM 25114 and a heavy chain sequence as deposited by way of plasmid deposition with deposit number DSM 25115, 
 iv. a RAM9 antibody, characterized by a light chain sequence as deposited by way of plasmid deposition with deposit number DSM 25859 and a heavy chain sequence as deposited by way of plasmid deposition with deposit number DSM 25860, 
 v. a RAM4 antibody, characterized by a light chain sequence as deposited by way of plasmid deposition with deposit number DSM 25861 and a heavy chain sequence as deposited by way of plasmid deposition with deposit number DSM 25862, 
 vi. a RAM0 antibody, characterized by a light chain sequence as deposited by way of plasmid deposition with deposit number DSM 25863 and a heavy chain sequence as deposited by way of plasmid deposition with deposit number DSM 25864, 
 vii. a RAB9 antibody, which is characterized by a light chain amino acid sequence of SEQ ID NO:22 and a heavy chain amino acid sequence of SEQ ID NO:26, 
 viii. a RAB4 antibody, which is characterized by a light chain amino acid sequence of SEQ ID NO:23 and a heavy chain amino acid sequence of SEQ ID NO:27, 
 ix. a RAB0 antibody, which is characterized by a light chain amino acid sequence of SEQ ID NO:24 and a heavy chain amino acid sequence of SEQ ID NO:28, 
 x. a RAM5 antibody, which is characterized by a light chain amino acid sequence of SEQ ID NO:33 and a heavy chain amino acid sequence of SEQ ID NO:32, 
 xi. a RAM4 antibody, which is characterized by a light chain amino acid sequence of SEQ ID NO:35 and a heavy chain amino acid sequence of SEQ ID NO:34, and 
 xii. a RAM0 antibody, which is characterized by a light chain amino acid sequence of SEQ ID NO:31 and a heavy chain amino acid sequence of SEQ ID NO:30. 
 
     
     
         14 . An essentially CHO MIF free anti-MIF antibody preparation as obtainable by use of the CHO-MIF knock-out CHO cell line according to  claim 8 . 
     
     
         15 . A method for the production of an essentially CHO MIF free anti-MIF antibody preparation which is characterized by the use of the CHO-MIF knock-out CHO cell line according to  claim 8 . 
     
     
         16 . A preparation of a recombinant MIF binding protein, preferably an (h) MIF binding protein, wherein said (h) MIF binding protein is selected from the group consisting of: an (h)MIF binding peptide, ligand, agonist, antagonist, inhibitor, or a MIF receptor fragment, and an anti-(h)MIF antibody preparation, and wherein said preparation is produced in a CHO cell line, characterized in that said preparation is essentially free of CHO-MIF. 
     
     
         17 . A preparation of a recombinant MIF binding protein, preferably an (h) MIF binding protein, wherein said (h) MIF binding protein is selected from the group consisting of: an (h)MIF binding peptide, ligand, agonist, antagonist, inhibitor, or a MIF receptor fragment, and an anti-(h)MIF antibody preparation, and wherein said preparation is obtainable by the method according to  claim 15 . 
     
     
         18 . A preparation of a recombinant MIF binding protein, preferably an (h)MIF binding protein, like an (h)MIF binding peptide, ligand, agonist, antagonist, inhibitor, or a MIF receptor fragment, or an anti-(h)MIF antibody preparation, produced by the method according to  claim 15 . 
     
     
         19 . The anti-hMIF antibody preparation of  claim 16 , which is essentially free of CHO MIF, wherein the anti-hMIF antibody is selected from the group consisting of:
 i. a RAB9 antibody, characterized by a light chain sequence as deposited by way of plasmid deposition with deposit number DSM 25111 and a heavy chain sequence as deposited by way of plasmid deposition with deposit number DSM 25113,   ii. a RAB4 antibody, characterized by a light chain sequence as deposited by way of plasmid deposition with deposit number DSM 25110 and a heavy chain sequence as deposited by way of plasmid deposition with deposit number DSM 25112,   iii. a RAB0 antibody, characterized by a light chain sequence as deposited by way of plasmid deposition with deposit number DSM 25114 and a heavy chain sequence as deposited by way of plasmid deposition with deposit number DSM 25115,   iv. a RAM9 antibody, characterized by a light chain sequence as deposited by way of plasmid deposition with deposit number DSM 25859 and a heavy chain sequence as deposited by way of plasmid deposition with deposit number DSM 25860,   v. a RAM4 antibody, characterized by a light chain sequence as deposited by way of plasmid deposition with deposit number DSM 25861 and a heavy chain sequence as deposited by way of plasmid deposition with deposit number DSM 25862,   vi. a RAM0 antibody, characterized by a light chain sequence as deposited by way of plasmid deposition with deposit number DSM 25863 and a heavy chain sequence as deposited by way of plasmid deposition with deposit number DSM 25864,   vii. a RAB9 antibody, which is characterized by a light chain amino acid sequence of SEQ ID NO:22 and a heavy chain amino acid sequence of SEQ ID NO:26,   viii. a RAB4 antibody, which is characterized by a light chain amino acid sequence of SEQ ID NO:23 and a heavy chain amino acid sequence of SEQ ID NO:27,   ix. a RAB0 antibody, which is characterized by a light chain amino acid sequence of SEQ ID NO:24 and a heavy chain amino acid sequence of SEQ ID NO:28,   x. a RAM9 antibody, which is characterized by a light chain amino acid sequence of SEQ ID NO:33 and a heavy chain amino acid sequence of SEQ ID NO:32,   xi. a RAM4 antibody, which is characterized by a light chain amino acid sequence of SEQ ID NO:35 and a heavy chain amino acid sequence of SEQ ID NO:34, and   xii. a RAM0 antibody, which is characterized by a light chain amino acid sequence of SEQ ID NO:31 and a heavy chain amino acid sequence of SEQ ID NO:30.