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|Title:||DNA ligands of thrombin|
|Abstract:||Methods are described for the identification of nucleic acid ligand solutions to thrombin. The present invention utilizes the SELEX (Systematic Evolution of Ligands for EXponential Enrichment) method for identfying and preparing DNA ligands to thrombin. Further included in the present invention are modified nucleotide sequences based on the sequences of the DNA ligands identified. The modified DNA ligands to thrombin exhibit increased in vivo stability.|
|Inventor(s):||Gold; Larry (Boulder, CO), Tasset; Diane (Boulder, CO)|
|Assignee:||NeXstar Pharmaceuticals, Inc. (Boulder, CO)|
|Filing Date:||Mar 28, 1994|
|Claims:||1. A method for identifying nucleic acid ligands to thrombin comprising: |
(a) preparing a candidate mixture of DNA nucleic acids;
(b) contacting the candidate mixture with thrombin, wherein nucleic acid ligands having an increased affinity to thrombin may be partitioned from the remainder of the candidate mixture;
c) partitioning between members of said candidate mixture on the basis of affinity to thrombin; and
d) amplifying partitioned members of the candidate mixture with a relatively higher affinity for thrombin compared to the candidate mixture as a whole, to yield a mixture of nucleic acids enriched for sequences with a higher affinity to thrombin, whereby nucleic acid ligands of thrombin may be identified.
2. A DNA nucleic acid ligand to thrombin identified according to the method of claim 1.
3. The nucleic acid ligand of claim 2 being a single stranded nucleic acid.
4. A purified and isolated non-naturally occurring DNA ligand to thrombin wherein said ligand is selected from the group consisting of the sequences set forth in FIGS. 9 and 10.
5. The method of claim 1 further including:
e) repeating steps b), c) and d).
6. The method of claim 5 wherein step e) is repeated at least 6 times.
7. A DNA nucleic acid ligand to thrombin identified according to the method of claim 5.
8. A DNA nucleic acid ligand to thrombin identified according to the method of claim 6.