Vascular endothelial cell growth factor variants and uses thereof

Abstract

The present invention provides VEGF variants having at least a single amino acid mutation in the native VEGF sequence and selective binding affinity for either the KDR receptor or the FLT-1 receptor. Methods of making the VEGF variants and methods of using the VEGF variants are also provided.

1 . A VEGF variant polypeptide comprising one or more amino acid mutations in the amino acid sequence of native VEGF and having selective binding affinity for KDR receptor.  
     
     
         2 . The VEGF variant of  claim 1 , wherein said one or more amino acid mutations comprises one or more amino acid substitutions at or between positions 17 to 25 of the native VEGF sequence.  
     
     
         3 . The VEGF variant of  claim 1 , wherein said one or more amino acid mutations comprises one or more amino acid substitutions at or between positions 63 to 66 of the native VEGF sequence.  
     
     
         4 . The VEGF variant of  claim 1 , wherein said polypeptide comprises at least four different amino acid mutations and four of said amino acid mutations are the following amino acid substitutions: M18E, Y21L, Q22R, Y25S.  
     
     
         5 . The VEGF variant of  claim 1 , wherein said polypeptide comprises at least three different amino acid mutations and three of said amino acid mutations are the following amino acid substitutions: D63S, G65M, L66R.  
     
     
         6 . The VEGF variant of  claim 1 , wherein said polypeptide comprises at least four different amino acid mutations and four of said amino acid mutations are the following amino acid substitutions: M18E, D63S, G65M, L66R.  
     
     
         7 . The VEGF variant of  claim 1 , wherein said polypeptide comprises at least four different amino acid mutations and four of said amino acid mutations are the following amino acid substitutions: Y21L, D63S, G65M, L66R.  
     
     
         8 . An isolated nucleic acid comprising a DNA sequence encoding the VEGF variant of  claim 1 .  
     
     
         9 . The isolated nucleic acid of  claim 8 , wherein said DNA sequence encodes the VEGF variant of  claim 4 .  
     
     
         10 . The isolated nucleic acid of  claim 8 , wherein said DNA sequence encodes the VEGF variant of  claim 5 .  
     
     
         11 . The isolated nucleic acid of  claim 8 , wherein said DNA sequence encodes the VEGF variant of  claim 6 .  
     
     
         12 . The isolated nucleic acid of  claim 8 , wherein said DNA sequence encodes the VEGF variant of  claim 7 .  
     
     
         13 . A vector comprising the nucleic acid of  claim 8 .  
     
     
         14 . A host cell comprising the vector of  claim 13 .  
     
     
         15 . A composition comprising the VEGF variant of  claim 1  and a carrier.  
     
     
         16 . The composition of  claim 15  wherein said carrier is a pharmaceutically acceptable carrier.  
     
     
         17 . An assay to detect KDR receptor, comprising contacting a cell or tissue with the VEGF variant of  claim 1  and detecting binding of said VEGF variant to KDR receptor(s) which may be present in or on said cell or tissue.  
     
     
         18 . A method of stimulating vasculogenesis or angiogenesis, comprising exposing mammalian cells expressing KDR receptor to an effective amount of the VEGF variant of  claim 1 .  
     
     
         19 . An article of manufacture, comprising a container holding the composition of  claim 15  and a label on said container providing instructions for use of said composition in vitro or in vivo.