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Details for Patent: 5,837,456

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Details for Patent: 5,837,456

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:08/488,402
Claims:1. A method of identifying nucleic acid ligands to a Chorionic Gonadotropin (CG)-related glycoprotein hormone, 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 acids with relatively higher affinity and specificity for binding to the CG-related glycoprotein hormone, whereby nucleic acid ligands of CG-related glycoprotein hormone may be identified.

2. The method of claim 1 further comprising:

e) repeating steps b), c), and d).

3. The method of claim 1 further comprising 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.

4. The method of claim 3 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.

5. A method of identifying nucleic acid ligands to a human Chorionic Gonadotropin (hCG)-related glycoprotein hormone, 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 acids with relatively higher affinity and specificity for binding to the hCG-related glycoprotein hormone, whereby nucleic acid ligands of hCG-related glycoprotein hormone may be identified.

6. The method of claim 5 further comprising:

e) repeating steps b), c), and d).

7. The method of claim 5 further comprising 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.

8. The method of claim 7 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.

9. The method of claim 5 wherein said candidate mixture of nucleic acids is comprised of single stranded nucleic acids.

10. The method of claim 9 wherein said single stranded nucleic acids are ribonucleic acids.

11. The method of claim 10 wherein said nucleic acids are 2'-amino (2'-NH.sub.2) modified ribonucleic acids.

12. The method of claim 10 wherein said nucleic acids are 2'-F modified ribonucleic acids.

13. The method of claim 5, wherein said hCG-related glycoprotein hormone is hCG.

14. The method of claim 5, wherein said hCG-related glycoprotein hormone is human thyroid-stimulating hormone (hTSH).
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