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|Title:||High affinity HKGF nucleic acid ligands and inhibitors|
|Abstract:||Methods are described for the identification and preparation of high-affinity nucleic acid ligands to hKGF. Included in the invention are specific RNA ligands to hKGF identified by the SELEX method. Also included are RNA ligands that inhibit the interaction of hKGF with its receptor.|
|Inventor(s):||Pagratis; Nikos (Boulder, CO), Gold; Larry (Boulder, CO)|
|Assignee:||Nexstar Pharmaceuticals, Inc. (Boulder, CO)|
|Filing Date:||Jun 05, 1995|
|Claims:||1. A method of identifying nucleic acid ligands to Human Keratinocyte Growth Factor (hKGF), comprising: |
a) preparing a candidate mixture of nucleic acids;
b) contacting the candidate mixture of nucleic acids with hKGF, wherein nucleic acids having an increased affinity to hKGF relative to the candidate mixture may be partitioned from the remainder of the candidate mixture;
c) partitioning the increased affinity nucleic acids from the remainder of the candidate mixture; and
d) amplifying the increased affinity nucleic acids to yield a mixture of nucleic acids enriched for nucleic acid sequences with relatively higher affinity and specificity for binding to hKGF, whereby nucleic acid ligands of hKGF may be identified.
2. The method of claim 1 further comprising:
e) repeating steps b), c), and d).
3. The method of claim 1 wherein said candidate mixture of nucleic acids is comprised of single stranded nucleic acids.
4. The method of claim 3 wherein said single stranded nucleic acids are ribonucleic acids.
5. The method of claim 4 wherein said nucleic acids are 2'-amine (2'--NH.sub.2) modified ribonucleic acids.
6. The method of claim 4 wherein said nucleic acids are 2'--F modified ribonucleic acids.