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|Title:||High-affinity RNA ligands of basic fibroblast growth factor|
|Abstract:||Methods are described for the identification and preparation of nucleic acid ligands and ligand solutions to basic fibroblast growth factor (bFGF). Included in the invention are nucleic acid ligands to bFGF which are inhibitors of bFGF and 2'-amino-modified RNA ligands to bFGF.|
|Inventor(s):||Janjic; Nebojsa (Boulder, CO), Gold; Larry (Boulder, CO)|
|Assignee:||NeXstar Pharmaceuticals, Inc. (Boulder, CO)|
|Filing Date:||Feb 10, 1994|
|Claims:||1. A method for identifying nucleic acid ligands to basic fibroblast growth factor (bFGF) comprising: |
a) preparing a candidate mixture of nucleic acids,
b) contacting the candidate mixture with bFGF, wherein nucleic acids having sequences with an increased affinity to bFGF relative to the total candidate mixture may be partitioned from the remainder of the nucleic acids in 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 sequences with increased affinity to bFGF, whereby nucleic acid ligands to bFGF may be identified.
2. The method of claim 1 further comprising e) repeating steps b) through 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 candidate mixture of nucleic acids is comprised of RNA.
5. The method of claim 4 wherein said candidate mixture of nucleic acids is comprised of RNA wherein all pyrimidines are 2'-deoxy-2-NH.sub.2 pyrimidines.
6. A method for selecting a nucleic acid ligand which inhibits the binding of bFGF to its receptor, which comprises the additional step of screening nucleic acid ligands to bFGF identified by the method of claim 1 for their ability to inhibit the binding of bFGF to its receptor.