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Details for Patent: 6,331,394

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Details for Patent: 6,331,394

Title: Nucleic acid ligands to integrins
Abstract:Methods are described for the isolation of nucleic acid ligands to integrins using the SELEX process. SELEX is an acronym for Systematic Evolution of Ligands by EXponential enrichment. The nucleic acid ligands of the present invention are useful as therapeutic and diagnostic agents.
Inventor(s): Ruckman; Judy (Boulder, CO), Gold; Larry (Boulder, CO), Stephens; Andrew (Boulder, CO), Janjic; Nebojsa (Boulder, CO)
Assignee: Gilead Sciences, Inc. (Foster City, CA)
Filing Date:Jul 29, 1999
Application Number:09/364,543
Claims:1. A nucleic acid ligand to an integrin identified according to the method comprising:

a) preparing a candidate mixture of nucleic acids;

b) contacting the candidate mixture of nucleic acids with said integrin, wherein nucleic acids having an increased affinity to said integrin 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;

d) amplifying the increased affinity nucleic acids to yield a mixture of nucleic acids enriched for nucleic acids with relatively higher affinity and specificity for binding to said integrin, whereby a nucleic acid ligand of said integrin may be identified.

2. The nucleic acid ligand of claim 1 wherein said integrin is a .beta..sub.3 integrin.

3. The nucleic acid ligand of claim 2 wherein said .crclbar..sub.3 integrin is selected from the group consisting of .alpha..sub.IIb.beta..sub.3 integrin and .alpha..sub.v.beta..sub.3 integrin.

4. A purified and isolated non-naturally occurring nucleic acid ligand to an integrin.

5. A purified and non-naturally occurring RNA ligand to an integrin wherein said ligand is selected from the group consisting of SEQ ID NO:5-118.

6. The nucleic acid ligand of claim 1 wherein said integrin is attached through either covalent or non-covalent bonds to a solid support, and wherein steps b)-c) take place on the surface of said solid support.

7. The nucleic acid ligand of claim 6 wherein said solid support is a bead.

8. The nucleic acid ligand of claim 1 wherein said candidate mixture of nucleic acids is comprised of single stranded nucleic acids.

9. The nucleic acid ligand of claim 8 wherein said single stranded nucleic acids are ribonucleic acids.

10. The nucleic acid ligand of claim 8 wherein said single stranded nucleic acids are deoxyribonucleic acids.

11. The nucleic acid ligand of claim 9 wherein said candidate mixture of nucleic acids comprises 2'-F(2'-fluoro) modified ribonucleic acids.

12. The purified and isolated non-naturally occurring nucleic acid ligand of claim 4 wherein said nucleic acid ligand is single stranded.

13. The purified and isolated non-naturally occurring nucleic acid ligand of claim 12 wherein said nucleic acid ligand is RNA.

14. The purified and isolated non-naturally occurring RNA ligand of claim 13 wherein said ligand is comprised of 2'-fluoro (2'-F) modified nucleotides.

15. A method for the isolation of nucleic acid ligands to an integrin, comprising:

a) preparing a candidate mixture of nucleic acids;

b) contacting the candidate mixture of nucleic acids with said integrin, wherein nucleic acids having an increased affinity to said integrin 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;

d) amplifying the increased affinity nucleic acids to yield a mixture of nucleic acids enriched for nucleic acids with relatively higher affinity and specificity for binding to said integrin, whereby a nucleic acid ligand of said integrin may be isolated; and

e) isolating nucleic acid ligands to an integrin.

16. The method of claim 15 wherein said candidate mixture comprises single-stranded nucleic acids.

17. The method of claim 16 wherein said single-stranded nucleic acids comprise ribonucleic acids.

18. The method of claim 15 wherein said integrin is a .beta..sub.3 integrin.

19. The method of claim 18 wherein said .beta..sub.3 integrin is .alpha..sub.IIb.beta..sub.3 integrin.
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