Tumor necrosis factor inhibitory protein and its purification

Abstract

Recombinant DNA encoding Tumor Necrosis Factor (TNF) Inhibitory Protein, or an active fragment thereof, is obtained. The TNF Inhibitory Protein has the ability to inhibit: (a) the binding of TNF to its receptors, and (b) the cytotoxic effect of TNF. TNF Inhibitory Protein and salts, functional derivatives and active fractions thereof can be used to antagonize the deleterious effects of TNF by eliminating TNF from the body, and to reduce the cytotoxic activity of TNF by binding to TNF and thereby to inhibit the binding of TNF to its receptors. The DNA may be in an expression vector. Host cells transfected with such an expression vector may be used to produce the TNF Inhibitory Protein.

What is claimed is:  
     
         1 . A recombinant DNA molecule comprising the nucleotide sequence encoding a polypeptide comprising the amino acid sequence: Asp-Ser-Val-Cys-Pro-Gln-Gly-Lys-Tyr-Ile-His-Pro-Gln-X-Asn-Ser (SEQ ID NO:1) wherein X is an unidentified amino acid residue, and said polypeptide has the ability to interact with TNF in such a manner as to: 
 (a) inhibit the binding of TNF to a TNF receptor; and    (b) inhibit the cytotoxic effect of TNF.    
     
     
         2 . A DNA molecule in accordance with  claim 1 , wherein the DNA molecule is genomic DNA or cDNA.  
     
     
         3 . A recombinant DNA molecule comprising: 
 (a) a nucleotide sequence encoding a polypeptide which comprises the amino acid sequence: Asp-Ser-Val-Cys-Pro-Gln-Gly-Lys-Tyr-Ile-His-Pro-Gln-X-Asn-Ser (SEQ ID NO:1)    wherein X is an unidentified amino acid residue and said polypeptide has the ability to interact with TNF in such a manner as to inhibit the binding of TNF to a TNF receptor and to inhibit the cytotoxic effect of TNF, or    (b) a nucleotide sequence encoding a fragment of said polypeptide, wherein said fragment has the ability to interact with TNF in such a manner as to inhibit the binding of TNF to a TNF receptor and to inhibit the cytotoxic effect of TNF.    
     
     
         4 . An expression vector comprising a DNA molecule in accordance with  claim 1   
     
     
         5 . An expression vector comprising a DNA molecule in accordance with  claim 3 .  
     
     
         6 . A host cell comprising an expression vector in accordance with  claim 4 .  
     
     
         7 . A host cell comprising an expression vector in accordance with  claim 7 .  
     
     
         8 . A method of producing a polypeptide capable of interacting with TNF, comprising culturing a host cell in accordance with  claim 6  and recovering the polypeptide produced thereby which is capable of interacting with TNF.  
     
     
         9 . A method of producing a polypeptide capable of interacting with TNF, comprising culturing a host cell in accordance with  claim 7  and recovering the polypeptide produced thereby which is capable of interacting with TNF.