Soluble recombinant botulinum toxins having a C-terminal portion of a heavy chain, a N-terminal portion of a heavy chain and a light chain

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 - 24 . (canceled)  
     
     
         25 . A recombinant botulinum toxin comprising a soluble C-terminal portion of a botulinum toxin heavy chain, an N-terminal portion of a botulinum toxin heavy chain and a botulinum toxin light chain 25 wherein the C-terminal portion comprises SEQ ID NO: 23 and the N-terminal portion and the light chain each comprise a portion of SEQ ID NO: 28.  
     
     
         26 . The recombinant toxin of  claim 25  further comprising a non-toxin protein sequence.  
     
     
         27 . The recombinant toxin of  claim 25  wherein the toxin is in a solution.  
     
     
         28 . The recombinant botulinum toxin of  claim 25  wherein the light chain is solubilized.  
     
     
         29 . The recombinant toxin of  claim 25  wherein the C-terminal portion and the N-terminal portion are bonded together.  
     
     
         30 . The recombinant toxin of  claim 29  wherein the bond is a covalent bond.  
     
     
         31 . The recombinant toxin of  claim 25  wherein the light chain is bonded to the C-terminal portion or the N-terminal portion.  
     
     
         32 . A recombinant botulinum toxin comprising a soluble C-terminal portion of a botulinum toxin type A heavy chain, an N-terminal portion of a botulinum toxin type A heavy chain and a botulinum toxin type A light chain wherein the C-terminal portion comprises SEQ ID NO: 23 and the N-terminal portion and the light chain each comprise a portion of SEQ ID NO: 28.  
     
     
         33 . The recombinant toxin of  claim 32  further comprising a non-toxin protein sequence.  
     
     
         34 . The recombinant toxin of  claim 32  wherein the toxin is in a solution.  
     
     
         35 . The recombinant botulinum toxin of  claim 32  wherein the light chain is solubilized.  
     
     
         36 . The recombinant toxin of  claim 32  wherein the C-terminal portion and the N-terminal portion are bonded together.  
     
     
         37 . The recombinant toxin of  claim 36  wherein the bond is a covalent bond.  
     
     
         38 . The recombinant toxin of  claim 32  wherein the light chain is bonded to the C-terminal portion or the N-terminal portion.