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Bivalent binding molecules of 7 transmembrane G protein-coupled receptors

  • US 6,682,886 B1
  • Filed: 07/17/1998
  • Issued: 01/27/2004
  • Est. Priority Date: 04/28/1994
  • Status: Expired due to Fees
First Claim
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1. A method of identifying a bivalent binding molecule of a 7TM G protein-coupled receptor wherein said bivalent binding molecule comprises a first and second aptamer to a first and second epitope of the same 7 transmembrane G protein-coupled receptor, said method comprising:

  • a) identifying said first aptamer to said first epitope by the method comprising;

    i) preparing a first candidate mixture of nucleic acids;

    ii) contacting said first candidate mixture of nucleic acid with said first epitope, wherein nucleic acids having an increased affinity to said first epitope may be partitioned from the remainder of the first candidate mixture;

    iii) partitioning said increased affinity nucleic acids from the remainder of the first candidate mixture; and

    iv) amplifying said increased affinity nucleic acids, whereby said first aptamer to said first epitope may be identified;

    b) identifying said second aptamer to said second epitope by the method comprising;

    i) preparing a second candidate mixture of nucleic acids;

    ii) contacting said second candidate mixture of nucleic acid with said second epitope, wherein nucleic acids having an increased affinity to said second epitope may be partitioned from the remainder of the second candidate mixture;

    iii) partitioning said increased affinity nucleic acids from the remainder of the second candidate mixture; and

    iv) amplifying said increased affinity nucleic acids, whereby said second aptamer to said second epitope may be identified; and

    c) coupling said first aptamer to said second aptamer, whereby said bivalent binding molecule may be identified;

    wherein said 7 transmembrane G protein-coupled receptor is selected from the group consisting of histamine, muscarinic, cholecystokinin, choriogonadotropin, corticotropin releasing factor, gastrin, neuropeptide Y, TSH and 5-HT 7 transmembrane G protein-coupled receptors.

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