Recombinant human endothelial cell growth factor

Abstract

Endothelial cell growth factor is achieved through the application of recombinant DNA technology to prepare cloning vehicles encoding the ECGF protein and procedures are disclosed for recovering ECGF protein essentially free of other proteins of human origin. The product is useful for, among other purposes, diagnostic applications and as potential in the treatment of damaged blood vessels or other endothelial cell-lined structures.

What is claimed is:  
     
       1. Purified and isolated biologically active human recombinant endothelial cell growth factor, said factor having an amino acid sequence selected from the group consisting of: 
       
         
           
                 
                 
                 
               
                     
                   (i) 
                   AEGETTFTALTEKFNLPPGNYKKPKLLYCSNGGHF 
                 
                     
                     
                 
                     
                     
                   LRILPDGTVDGTRDRSDQHIQLQSAESVGEVYIKST 
                 
                     
                     
                 
                     
                     
                   ETGQYLAMDTDGLLYGSQTPNEECLFLERLEENHYN 
                 
                     
                     
                 
                     
                     
                   TYISKKHAEKNWFYGLKKNGSCKRGPRTHYGQKAIL 
                 
                     
                     
                 
                     
                     
                   FLPLPYSSD, and 
                 
             
                
                
                
                
                
                
                
                
                
               
            
           
         
       
       (ii) a sequence of (i) additionally comprising an NH 2 -terminal methionine residue,  
       said factor being free of other proteins of human origin. 
     
     
       2. The factor according to  claim 1  in admixture with an acceptable carrier. 
     
     
       3. Purified and isolated biologically active human recombinant endothelial cell growth factor, said factor having an amino acid sequence selected from the group consisting of: 
       
         
           
                 
                 
                 
               
                     
                   (i) 
                   NYKKPLLYCSNGGHFLRILPDGTVDGTRDRSDQHI 
                 
                     
                     
                 
                     
                     
                   QLQLSAESVGEVYIKSTETGQYLAMDTDGLLYGSQ 
                 
                     
                     
                 
                     
                     
                   TPNEECLFLERLEENHYNTYISKKHAEKNWFVGLKK 
                 
                     
                     
                 
                     
                     
                   NGSCKRGPRTHYGQKAILFLPLPVSSD; and 
                 
             
                
                
                
                
                
                
                
               
            
           
         
       
       (ii) a sequence of (i) additionally comprising an NH 2 -terminal methionine residue,  
       said factor being free of other proteins of human origin. 
     
     
       4. The factor according to  claim 3  in admixture with an acceptable carrier. 
     
     
       5. An isolated and purified biologically active human endothelial cell growth factor produced by culturing a prokaryotic or eukaryotic host cell containing a DNA having a sequence comprising: 
       
         
           
                 
                 
               
                     
                   GCT GAA GGG GAA ATC ACC ACC TTC ACA GCC 
                 
                     
                     
                 
                     
                   CTG ACC GAG AAG TTT AAT CTG CCT CCA GGG 
                 
                     
                     
                 
                     
                   AAT TAC AAG AAG CCC AAA CTC CTC TAC TGT 
                 
                     
                     
                 
                     
                   AGC AAC GGG GGC CAC TTC CTG AGG ATC CTT 
                 
                     
                     
                 
                     
                   CCG GAT GGC ACA GTG GAT GGG ACA AGG GAC 
                 
                     
                     
                 
                     
                   AGG AGC GAC CAG CAC ATT CAG CTG CAG CTC 
                 
                     
                     
                 
                     
                   AGT GCG GAA AGC GTG GGG GAG GTG TAT ATA 
                 
                     
                     
                 
                     
                   AAG AGT ACC GAG ACT GGC CAG TAC TTG GCC 
                 
                     
                     
                 
                     
                   ATG GAC ACC GAC GGG CTT TTA TAC GGC TCA 
                 
                     
                     
                 
                     
                   CAG ACA CCA AAT GAG GAA TGT TTG TTC CTG 
                 
                     
                     
                 
                     
                   GAA AGG CTG GAG GAG AAC CAT TAC AAC ACC 
                 
                     
                     
                 
                     
                   TAT ATA TCC AAG AAG CAT GCA GAG AAG AAT 
                 
                     
                     
                 
                     
                   TGG TTT GTT GGC CTC AAG AAG AAT GGG AGC 
                 
                     
                     
                 
                     
                   TGC AAA CGC GGT CCT CGG ACT CAC TAT GGC 
                 
                     
                     
                 
                     
                   CAG AAA GCA ATC TTG TTT CTC CCC CTG CCA 
                 
                     
                     
                 
                     
                   GTC TCT TCT GAT. 
                 
             
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
               
            
           
         
       
     
     
       6. The protein  endothelial cell growth factor according to  claim 5 , wherein said protein  endothelial cell growth factor has been purified from said host cell. 
     
     
       7. The protein  endothelial cell growth factor according to  claim 5  in admixture with an acceptable carrier. 
     
     
       8. The protein  endothelial cell growth factor according to  claim 5  wherein said protein  endothelial cell growth factor produced by culturing a prokaryotic or eukaryotic host cell containing said DNA is recovered by reaction of the protein  endothelial cell growth factor expressed by the host cell with a reagent composition comprising at least one binding protein specific for endothelial cell growth factor. 
     
     
       9. An isolated and purified biologically active human endothelial cell growth factor produced by culturing a prokaryotic or eukaryotic host cell containing a DNA having a sequence comprising: 
       
         
           
                 
                 
               
                     
                   AAT TAC AAG AAG CCC AAA CTC CTC TAC TGT 
                 
                     
                     
                 
                     
                   AGC AAC GGG GGC CAC TTC CTG AGG ATC CTT 
                 
                     
                     
                 
                     
                   CCG GAT GGC ACA GTG GAT GGG ACA AGG GAC 
                 
                     
                     
                 
                     
                   AGG AGC GAC CAG CAC ATT CAG CTG CAG CTC 
                 
                     
                     
                 
                     
                   AGT GCG GAA AGC GTG GGG GAG GTG TAT ATA 
                 
                     
                     
                 
                     
                   AAG AGT ACC GAG ACT GGC CAG TAC TTG GCC 
                 
                     
                     
                 
                     
                   ATG GAC ACC GAC GGG CTT TTA TAC GGC TCA 
                 
                     
                     
                 
                     
                   CAG ACA CCA AAT GAG GAA TGT TTG TTC CTG 
                 
                     
                     
                 
                     
                   GAA AGG CTG GAG GAG AAC CAT TAC AAC ACC 
                 
                     
                     
                 
                     
                   TAT ATA TCC AAG AAG CAT GCA GAG AAG AAT 
                 
                     
                     
                 
                     
                   TGG TTT GTT GGC CTC AAG AAG AAT GGG AGC 
                 
                     
                     
                 
                     
                   TGC AAA CGC GGT CCT CGG ACT CAC TAT GGC 
                 
                     
                     
                 
                     
                   CAG AAA GCA ATC TTG TTT CTC CCC CTG CCA 
                 
                     
                     
                 
                     
                   GTC TCT TCT GAT. 
                 
             
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
               
            
           
         
       
     
     
       10. The protein  endothelial cell growth factor according to  claim 9 , wherein said protein  endothelial cell growth factor has been purified from said host cell. 
     
     
       11. The protein  endothelial cell growth factor according to  claim 9  in admixture with an acceptable carrier. 
     
     
       12. The protein  endothelial cell growth factor according to  claim 9  wherein said protein  endothelial cell growth factor produced by culturing a prokaryotic or eukaryotic host cell containing said DNA is recovered by reaction of the protein  endothelial cell growth factor expressed by the host cell with a reagent composition comprising at least one binding protein specific for endothelial cell growth factor.