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|Title:||Systematic evolution of ligands exponential enrichment: blended selex|
|Abstract:||A method is described for generating blended nucleic acid ligands containing non-nucleic acid functional units. Specifically, a SELEX identified RNA ligand to the integrin gpIIbIIIa is conjugated to the peptide Gly-Arg-Gly-Asp-Thr-Pro (SEQ ID NO:1). This blended RNA ligand inhibits the biological activity of gpIIbIIIa with high specificity. Also described is a single-stranded DNA ligand to elastase coupled to N-methoxysuccinyl-Ala-Ala-Pro-Val-chloromethyl (SEQ ID NO:3) ketone. This elastase blended nucleic acid ligand inhibits the biological activity of elastase.|
|Inventor(s):||Biesecker; Greg (Boulder, CO), Jayasena; Sumedha D. (Boulder, CO), Gold; Larry (Boulder, CO), Smith; Drew (Boulder, CO), Kirschenheuter; Gary (Arvada, CO)|
|Assignee:||NeXstar Pharmaceuticals, Inc. (Boulder, CO)|
|Filing Date:||Oct 23, 1997|
|Claims:||1. A method for identifying blended nucleic acid ligands of a target compound from a blended candidate mixture comprised of blended nucleic acids each having at least one nucleic acid region and at least one functional unit, said method comprising: |
a) contacting the blended candidate mixture with the target, wherein blended nucleic acids having an increased affinity to the target relative to the blended candidate mixture may be partitioned from the remainder of the blended candidate mixture;
b) partitioning the increased affinity blended nucleic acids from the remainder of the blended candidate mixture; and
c) amplifying the increased affinity blended nucleic acids to yield a ligand-enriched mixture of blended nucleic acids, whereby blended nucleic acid ligands of the target compound may be identified.
2. The method of claim 1 wherein said at least one nucleic acid region is composed of a fixed region and a randomized region.
3. The method of claim 2 wherein said at least one functional unit is attached to an oligonucleotide hybridized to said fixed region.
4. A blended nucleic acid ligand identified according to the method of claim 1.
5. A method for preparing blended nucleic acid ligands of a target compound comprising:
a) identifying a nucleic acid ligand of a target compound from a candidate mixture comprised of nucleic acids each having a region of randomized sequence by a method comprising: i) contacting the candidate mixture with the target, wherein nucleic acids having an increased affinity to the target relative to the candidate mixture may be partitioned from the remainder of the candidate mixture; ii) partitioning the increased affinity nucleic acids from the remainder of the candidate mixture, and iii) amplifying the increased affinity nucleic acids to yield a ligand-enriched mixture of nucleic acids, whereby nucleic acid ligands of the target may be identified; and
b) attaching at least one functional unit to said nucleic acid ligand to yield a blended nucleic acid ligand of the target compound.
6. A blended nucleic acid ligand prepared according to the method of claim 3.
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