Method for Producing Protein-Carbohydrate Vaccines Reduced in Free Carbohydrate

Abstract

This invention is directed to processes for reducing the level of free carbohydrate from a solution of protein-linked carbohydrate (conjugate) and non-linked carbohydrate. In this process, the conjugate is adsorbed to a hydrophobic membrane while the carbohydrate is not. The conjugate is then desorbed from the membrane, yielding a solution that is substantially reduced in free carbohydrate.

1 - 11 . (canceled) 
     
     
         12 . A conjugate composition containing a reduced amount of free carbohydrate made by a process comprising:
 mixing the conjugate composition with a lyotropic salt;   passing the mixture through a hydrophobic interaction membrane; and   eluting the conjugate with an eluent.   
     
     
         13 . The composition of  claim 12 , which is a vaccine. 
     
     
         14 - 21 . (canceled) 
     
     
         22 . A conjugate composition containing a reduced amount of free carbohydrate made by a process comprising:
 mixing the conjugate composition with a lyotropic salt;   passing the mixture through a monolith matrix or column; and   eluting the conjugate with an eluent.   
     
     
         23 . The conjugate of  claim 22 , wherein the conjugate is derived from  Haemophilus, Neisseria, Salmonella, Streptococcus  or Tetanus, CRM 197  or protein D. 
     
     
         24 . The conjugate of  claim 22 , which is a vaccine. 
     
     
         25 . The composition of  claim 12 , wherein the carbohydrate comprises one or more of high molecular weight and low molecular weight polysaccharides, oligosaccharides, carbohydrate polymers, monomeric and polymeric sugars and combinations thereof. 
     
     
         26 . The composition of  claim 12 , which is independent of the chemistry used to create the conjugate and independent of type of carrier protein used. 
     
     
         27 . The composition of  claim 12 , wherein the conjugate is eluted with an eluent containing a reduced concentration of the lyotropic salt. 
     
     
         28 . The composition of  claim 27 , wherein the lyotropic salt is ammonium sulfate or NaCl. 
     
     
         29 . The composition of  claim 12 , wherein the eluent further includes an additive. 
     
     
         30 . The composition of  claim 29 , wherein the additive is an organic solvent, a detergent, or both. 
     
     
         31 . The composition of  claim 29 , wherein the additive is ethylene glycol. 
     
     
         32 . The composition of  claim 12 , wherein eluting is performed as a step or continuous gradient. 
     
     
         33 . The composition of  claim 12 , wherein greater than 50% of free carbohydrate is removed from the conjugate composition. 
     
     
         34 . The composition of  claim 12 , wherein greater than 90% of free carbohydrate is removed from the conjugate composition. 
     
     
         35 . The composition of  claim 22 , wherein the monolith matrix or column is a phenyl monolith. 
     
     
         36 . The composition of  claim 22 , wherein the eluent further includes an additive. 
     
     
         37 . The composition of  claim 36 , wherein the additive is an organic solvent, a detergent, or both. 
     
     
         38 . The composition of  claim 36 , wherein the additive is ethylene glycol. 
     
     
         39 . The composition of  claim 22 , wherein the conjugate is a vaccine. 
     
     
         40 . The composition of  claim 22 , wherein greater than 50% of free carbohydrate is removed from the conjugate composition. 
     
     
         41 . The composition of  claim 22 , wherein greater than 90% of free carbohydrate is removed from the conjugate composition.