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|Title:||High affinity nucleic acid ligands of cytokines|
|Abstract:||Methods are described for the identification and preparation of high-affinity nucleic acid ligands to cytokines. Included in the invention are specific nucleic acid ligands to IFN-gamma, IL-4, IL-10, TNF-alpha, and RANTES.|
|Inventor(s):||Tasset; Diane (Boulder, CO), Pagratis; Nikos (Boulder, CO), Jayasena; Sumedha (Boulder, CO), Gold; Larry (Boulder, CO)|
|Assignee:||NeXstar Pharmaceuticals, Inc. (Boulder, CO)|
|Filing Date:||Jun 07, 1995|
|Claims:||1. A method of identifying nucleic acid ligands to a cytokine selected from the group consisting of interleukin-10 (IL-10), interleukin-4 (IL-4), tumor necrosis factor-alpha (TNF-alpha) and Regulated on Activation, Normal T-cell Expressed and Secreted (RANTES), comprising: |
a) preparing a candidate mixture of nucleic acids;
b) contacting the candidate mixture of nucleic acids with said cytokine, wherein nucleic acids having an increased affinity to said cytokine 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 said cytokine, whereby nucleic acid ligands of said cytokine 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'-amino (2'-NH.sub.2) modified ribonucleic acids.
6. The method of claim 4 wherein said nucleic acids are 2'-fluoro (2'-F) modified ribonucleic acids.
7. The method of claim 3 wherein said single stranded nucleic acids are deoxyribonucleic acids.
8. The method of claim 1 wherein said cytokine is IL-10.
9. The method of claim 1 wherein said cytokine is IL-4.
10. The method of claim 1 wherein said cytokine is TNF-alpha.
11. The method of claim 1 wherein said cytokine is RANTES.
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