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|Title:||High-affinity oligonucleotide ligands to secretory phospholipase A2 (sPLA.sub.2)|
|Abstract:||This invention discloses high-affinity oligonucleotide ligands to human secretory phospholipase A2 (sPLA.sub.2), specifically RNA ligands having the ability to bind to sPLA.sub.2, and the methods for obtaining such ligands.|
|Inventor(s):||Parma; David H. (Boulder, CO), Gold; Larry (Boulder, CO)|
|Assignee:||Nexstar Pharmaceuticals, Inc. (Boulder, CO)|
|Filing Date:||Mar 06, 1995|
|Claims:||1. A method for identifying nucleic acid ligands and nucleic acid ligand sequences to secretory phospholipase A2 (sPLA.sub.2) comprising: |
a) contacting a candidate mixture of nucleic acids with sPLA.sub.2, wherein nucleic acids having an increased affinity to the sPLA.sub.2 relative to the candidate mixture may be partitioned from the remainder of the candidate mixture;
b) partitioning the increased affinity nucleic acids from the remainder of the candidate mixture; and
c) 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 sPLA.sub.2, whereby nucleic acid ligands and nucleic acid ligand sequences of sPLA.sub.2 may be identified.
2. The method of claim 1 further comprising:
d) repeating steps a), b) and c).
3. The method of claim 1 wherein said candidate mixture 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 3 wherein said single-stranded nucleic acids are deoxyribonucleic acids.
6. A nucleic acid ligand to sPLA.sub.2 identified according to the method of claim 1.
7. A purified and isolated non-naturally occurring nucleic acid ligand to secretory phospholipase A.sub.2 (sPLA.sub.2).
8. The nucleic acid ligand of claim 7 which is a ribonucleic acid ligand.
9. The RNA ligand of claim 8 wherein said RNA ligand is selected from the group consisting of the nucleotide sequences set forth in Table II, SEQ ID NOS:10-122.
10. A purified and isolated non-naturally occuring RNA ligand to sPLA.sub.2 comprising the sequences selected from the group consisting of SEQ ID NOS: 97-100.
11. The ligand of claim 7 wherein said ligand comprises a chemically modified pyrimidine base selected from the group consisting of 2'- NH.sub.2 pyrimidines and 2'-F pyrimidines.