High affinity oligonucleotide ligands to growth factors

Abstract

Methods are described for the identification and preparation of high-affinity nucleic acid ligands to TGFβ, PDGF and hKGF. Included in the invention are specific RNA and ssDNA ligands to TGFβ1 and PDGF identified by the SELEX method. Also included in the invention are specific RNA ligands to hKGF identified by the SELEX method. Further included are RNA ligands that inhibit the interaction of TGFβ1 and hKGF with their receptors and DNA ligands that inhibit the interaction of PDGF with its receptor.

1 . A method of identifying modified single-stranded ribonucleic acid ligands to PDGF comprising: 
 a) contacting a candidate mixture of modified single-stranded ribonucleic acids with PDGF, wherein ribonucleic acids having an increased affinity to PDGF relative to the candidate mixture may be partitioned from the remainder of the candidate mixture;    b) partitioning the increased affinity ribonucleic acids from the remainder of the candidate mixture; and    c) amplifying the increased affinity ribonucleic acids to yield a mixture of nucleic acids enriched for nucleic acid sequences with relatively higher affinity and specificity for binding to PDGF, whereby modified ribonucleic acid ligands of PDGF may be identified.    
     
     
         2 . A method of identifying modified single-stranded ribonucleic acid ligands to hKGF comprising: 
 a) contacting a candidate mixture of modified single-stranded ribonucleic acids with hKGF, whereby ribonucleic acids having an increased affinity to hKGF relative to the candidate mixture may be partitioned from the remainder of the candidate mixture;    b) partitioning the increased affinity ribonucleic acids from the remainder of the candidate mixture; and    c) amplifying the increased affinity ribonucleic acids to yield a mixture of ribonucleic acids enriched for ribonucleic acid sequences with relatively higher affinity and specificity for binding to hKGF, whereby modified ribonucleic acid ligands of hKGF may be identified.