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US 5,660,985 AGranted
High affinity nucleic acid ligands containing modified nucleotides
AssigneeNeXstar Pharmaceuticals, Inc.
InventorsWolfgang Pieken, Diane Tasset, Nebojsa Janjic, Larry Gold, Gary P. Kirschenheuter
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Abstract
Nucleic acid ligands containing modified nucleotides are described as are methods for producing such oligonucleotides. Such ligands enrich the chemical diversity of the candidate mixture for the SELEX process. Specific examples are provided of nucleic acids containing nucleotides modified at the 2'- and 5-position. Specific 2-OH and 2'-NH.sub.2 modified RNA ligands to thrombin are described.
Claims10
A method for identifying nucleic acid ligands comprising a modified nucleotide to a target molecule comprising:
- a) preparing a candidate mixture of single-stranded nucleic acids wherein each nucleic acid of said candidate mixture comprises a modified nucleotide selected from the group consisting of a 5-position modified pyrimidine, a 2'-position modified pyrimidine, an 8 -position modified purine, and a 2'-position modified purine;
- b) contacting the candidate mixture with said target molecule;
- c) partitioning the nucleic acids having an increased affinity to the target molecule relative to the candidate mixture from the remainder of the candidate mixture;
- and
- d) amplifying the increased affinity nucleic acids, in vitro, to yield a ligand-enriched mixture of nucleic acids, whereby nucleic acid ligands of the target molecule are identified.
2 more elements
Dependencies are inferred from the claim text.