Crystalline anti-htnfalpha antibodies

Abstract

The present invention relates to a batch crystallization method for crystallizing an anti-hTNFalpha antibody which allows the production of said antibody on an industrial scale; antibody crystals as obtained according to said method; compositions containing said crystals as well as methods of use of said crystals and compositions.

1 . A batch crystallization method for crystallizing an anti-hTNFalpha antibody, comprising the steps of:
 (a) providing an aqueous solution of said antibody in admixture with an inorganic phosphate salt as crystallization agent; and   (b) incubating said aqueous crystallization mixture until crystals of said antibody are formed.   
     
     
         2 . The crystallization method according to  claim 1 , wherein said aqueous crystallization mixture
 a) has a pH in the range of about pH 3 to 5; or   b) comprises a buffer.   
     
     
         3 . (canceled) 
     
     
         4 . The crystallization method according to  claim 2 , wherein said buffer comprises an acetate buffer. 
     
     
         5 - 8 . (canceled) 
     
     
         9 . The crystallization method according to  claim 1 , wherein the phosphate salt is selected from the group consisting of
 a) a hydrogenphosphate salt;   b) an alkali metal salt; and   c) a mixture of at least two different alkali metal salts.   
     
     
         10 - 12 . (canceled) 
     
     
         13 . The crystallization method according to  claim 1 , wherein at least one of the following additional crystallization conditions are met: a) incubation is performed for between about 1 hour to about 60 days; b) incubation is performed at a temperature between about 4° C. and about 37° C.; c) the antibody concentration is in the range of about 0.5 to about 100 mg/ml. 
     
     
         14 . The crystallization method according to  claim 13 , further comprising the step of drying said crystals. 
     
     
         15 . (canceled) 
     
     
         16 . The crystallization method according to  claim 13 , wherein the batch volume is in the range of about 1 ml to 20.000 liters. 
     
     
         17 . A crystal of an anti-hTNFalpha antibody, obtainable by a crystallization method according to  claim 1 . 
     
     
         18 . A crystal of an anti-hTNFalpha antibody, with the proviso that said antibody is not INFLIXIMAB. 
     
     
         19 . The crystal of  claim 17  having a needle-like morphology with a maximum length l of about 2-500 μm and an l/d ratio of about 3 to 30. 
     
     
         20 . The crystal of  claim 18  having a needle-like morphology with a maximum length l of about 2-500 μm and an l/d ratio of about 3 to 30. 
     
     
         21 . The crystal according to  claim 17 , wherein said antibody is selected from a group consisting of
 a) a polyclonal antibody;   b) a monoclonal antibody;   c) a human antibody;   d) a humanized antibody;   e) a non-glycosylated antibody; and   f) a chimeric antibody.   
     
     
         22 - 25 . (canceled) 
     
     
         26 . The crystal according to  claim 18 , wherein the antibody is an isolated human antibody; selected from the group consisting of
 a) an isolated human antibody that dissociates from hTNFalpha with a Kd of 1×10 −8  M or less and a Ka rate constant of 1×10 −3  s −1  or less, both determined by surface plasmon resonance, and neutralizes hTNFalpha cytotoxicity in a standard in vitro L929 assay with an IC 50  of 1×10 −7  M or less;   b) an isolated human antibody with the following characteristics: i) dissociates from human TNFalpha with a Koff rate constant of 1×10-3 s-1 or less, as determined by surface plasmon resonance; ii) has a light chain CDR3 domain comprising the amino acid sequence of SEQ ID NO: 3, or modified from SEQ ID NO: 3 by a single alanine substitution at position 1, 4, 5, 7 or 8 or by one to five conservative amino acid substitutions at positions 1, 3, 4, 6, 7, 8 and/or 9: iii) has a heavy chain CDR3 domain comprising the amino acid sequence of SEQ ID NO: 4, or modified from SEQ ID NO: 4 by a single alanine substitution at position 2, 3, 4, 5, 6, 8, 9, 10 or 11 or by one to five conservative amino acid substitutions at positions 2, 3, 4, 5, 6, 8, 9, 10, 11 and/or 12;   c) an isolated human antibody with a light chain variable region (LCVR) comprising the amino acid sequence of SEQ ID NO: 1 and a heavy chain variable region (HCVR) comprising the amino acid sequence of SEQ ID NO: 2; and   d) the adalimumab.   
     
     
         27 - 29 . (canceled) 
     
     
         30 . A pharmaceutical composition comprising: (a) crystals of an anti-hTNFalpha antibody according to  claim 17 , and (b) at least one pharmaceutical excipient; which composition is provided in as solid, semisolid or liquid formulation, each formulation containing said antibody in crystalline form. 
     
     
         31 - 35 . (canceled) 
     
     
         36 . An injectable liquid composition comprising anti-hTNFalpha antibody crystals according to  claim 17  and having an antibody concentration in the range of about 10 to 400 mg/ml. 
     
     
         37 . A crystal slurry comprising anti-hTNFalpha antibody crystals according to  claim 17 , having a antibody concentration greater than about 100 mg/ml. 
     
     
         38 . A method for treating a mammal comprising the step of administering to the mammal an effective amount of antibody anti-hTNFalpha crystals according to  claim 17 . 
     
     
         39 . A method for treating a mammal comprising the step of administering to the mammal an effective amount of the composition according to  claim 30 . 
     
     
         40 . (canceled) 
     
     
         41 . A method of treating a TNFalpha-related disorder in a subject, which method comprises administering a therapeutically effective amount of antibody crystals according to  claim 17 . 
     
     
         42 - 44 . (canceled)