Soluble recombinant botulinum toxin proteins

Abstract

The present invention includes recombinant proteins derived from Clostridium botulinum toxins. In particular, soluble recombinant Clostridium botulinum type A, type B and type E toxin proteins are provided. Methods which allow for the isolation of recombinant proteins free of significant endotoxin contamination are provided. The soluble, endotoxin-free recombinant proteins are used as immunogens for the production of vaccines and antitoxins. These vaccines and antitoxins are useful in the treatment of humans and other animals at risk of intoxication with clostridial toxin.

1 - 49 . (canceled)  
     
     
         50 . A composition comprising an active soluble recombinant botulinum neurotoxin, which comprises a non-toxin protein sequence and a botulinum toxin type B protein sequence having proteolytic activity.  
     
     
         51 . The composition of  claim 50 , wherein the non-toxin protein sequence is a poly-histidine tract.  
     
     
         52 . The composition of  claim 50 , wherein the botulinum toxin type B protein sequence is SEQ ID NO:40.  
     
     
         53 . The composition of  claim 50 , which is substantially endotoxin free.  
     
     
         54 . A composition comprising an active soluble recombinant botulinum neurotoxin, which comprises a non-toxin protein sequence and a botulinum toxin type E protein sequence having proteolytic activity.  
     
     
         55 . The composition of  claim 54 , wherein the non-toxin protein sequence is a poly-histidine tract.  
     
     
         56 . The composition of  claim 54 , wherein the botulinum neurotoxin type E sequence is selected from the group consisting of SEQ ID NO:50 and SEQ ID NO:52.  
     
     
         57 . The composition of  claim 54 , which is substantially endotoxin free.  
     
     
         58 . A method for producing a soluble recombinant botulinum neurotoxin, comprising: 
 expressing an active botulinum neurotoxin from an expression vector comprising a botulinum neurotoxin nucleotide sequence that encodes a botulinum neurotoxin having a proteolytic activity, and a solubility tag.    
     
     
         59 . The method of  claim 58 , wherein the botulinum neurotoxin is selected from the group consisting of botulinum neurotoxin types B and E.  
     
     
         60 . The method of  claim 58 , wherein the solubility tag is a poly-histidine tract.  
     
     
         61 . The method of  claim 58 , wherein the botulinum neurotoxin has the amino acid sequence of SEQ ID NO:40.  
     
     
         62 . The method of  claim 58 , wherein the botulinum neurotoxin has an amino acid sequence selected from the group consisting of SEQ ID NO:50 and SEQ ID NO:52.