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Details for Patent: 5,849,890

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Details for Patent: 5,849,890

Title: High affinity oligonucleotide ligands to chorionic gonadotropin hormone and related glycoprotein hormones
Abstract:Methods are described for the identification and preparation of nucleic acid ligands to CG-related glycoprotein hormones. Included in the invention are specific RNA ligands to hCG and hTSH identified by the SELEX method.
Inventor(s): Gold; Larry (Boulder, CO), Jayasena; Sumedha D. (Boulder, CO), Nieuwlandt; Dan (Broomfield, CO), Davis; Ken (Los Altos, CA)
Assignee: NeXstar Pharmaceuticals, Inc. (Boulder, CO)
Filing Date:Jun 07, 1995
Application Number:08/484,552
Claims:1. A purified and isolated non-naturally occurring nucleic acid ligand to a chorionic gonadotropin (CG)-related glycoprotein hormone.

2. The nucleic acid ligand to a CG-related glycoprotein hormone of claim 1 identified according to the method comprising:

a) preparing a candidate mixture of nucleic acids;

b) contacting the candidate mixture of nucleic acids with a CG-related glycoprotein hormone, wherein nucleic acids having an increased affinity to the CG-related glycoprotein hormone 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 the CG-related glycoprotein hormone, whereby nucleic acid ligands of the CG-related glycoprotein hormone may be identified.

3. The nucleic acid ligand to a CG-related glycoprotein hormone of claim 2 wherein the method further comprises contacting the increased affinity nucleic acids with one or more non-target molecules, wherein nucleic acids with affinity to the non-target molecule(s) are removed, and wherein the non-target molecule is a member of the CG-related glycoprotein hormone family other than the one for which increased affinity nucleic acids are being selected.

4. A purified and isolated non-naturally occurring nucleic acid ligand to an hCG-related glycoprotein hormone.

5. The nucleic acid ligand to an hCG-related glycoprotein hormone of claim 4 identified according to the method comprising:

a) preparing a candidate mixture of nucleic acids;

b) contacting the candidate mixture of nucleic acids with an hCG-related glycoprotein hormone, wherein nucleic acids having an increased affinity to the hCG-related glycoprotein hormone 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 the hCG-related glycoprotein hormone, whereby nucleic acid ligands of the hCG-related glycoprotein hormone may be identified.

6. The nucleic acid ligand to an hCG-related glycoprotein hormone of claim 5 wherein the method further comprises contacting the increased affinity nucleic acids with one or more non-target molecules, wherein nucleic acids with affinity to the non-target molecule(s) are removed, and wherein the non-target molecule is a member of the hCG-related glycoprotein hormone family other than the one for which increased affinity nucleic acids are being selected.

7. The purified and isolated non-naturally occurring nucleic acid ligand of claim 4, wherein said hCG-related glycoprotein hormone is hCG, and wherein said ligand is an RNA selected from the group consisting of the sequences set forth in FIGS. 3 and 4 and Tables 2, 5, 11 and 13 (SEQ ID NOS:4-30, 34-35, 41, 80-169).

8. The purified and isolated non-naturally occurring nucleic acid ligand of claim 4, wherein said hCG-related glycoprotein hormone is hTSH, and wherein said ligand is an RNA selected from the group consisting of the sequences set forth in Table 7 (SEQ ID NOS:42-63).

9. The purified and isolated non-naturally occurring nucleic acid ligand of claim 5, wherein said hCG-related glycoprotein hormone is hCG.

10. The purified and isolated non-naturally occurring nucleic acid ligand of claim 5, wherein said hCG-related glycoprotein hormone is hTSH.
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