Patent ID: 6537763
Filing Date: 2003-03-25
Classification: A61K,A61P,C07K,C12Q

Abstract:
A method of screening a molecule for its capacity to agonize or antagonize RANK, said method comprising the steps of:a) incubating a RANK polypeptide in the presence of the molecule, wherein said RANK polypeptide is selected from the group consisting of: (i) a polypeptide comprising amino acids x to y of SEQ ID NO:6. wherein x is any one of amino acids 1-33 and y is any one of amino acids 213-616; (ii) a polypeptide comprising amino acids 33-213 of SEQ ID NQ:6; (iii) a polypeptide comprising amino adds 234-616 of SEQ ID NO:6; (iv) a RANK/GST fusion protein, wherein RANK comprises amino acids 33-213 of SEQ ID NO:6; (v) a RANK/Fc fusion protein, wherein RANK comprises amino acids 33-213 of SEQ ID NO:6; (vi) a polypeptide comprising amino acids 544-616 of SEQ ID NO:6; (vii) a polypeptide comprising amino acids 339-421 of SEQ ID NO:6; (viii) a polypeptide comprising an amino acid sequence that is at least 80% identical to a polypeptide of (ii) and that is capable of binding a RANKL polypeptide comprising amino acids 69-317 of SEQ ID NO:13; and (ix) a polypeptide that is capable of binding a RANKL polypeptide comprising amino acids 69-317 of SEQ ID NO:13 and that is further capable of being encoded by a nucleic acid molecule that hybridizes under stringent conditions to the complement of a DNA having a nucleotide sequence as shown in SEQ ID NO:5, wherein said stringent conditions comprise hybridizing in 6Ã—SSC at 63Â° C. and washing in 3Ã—SSC at 55Â° C.; b)measuring a biological activity associated with the RANK polypeptide, wherein said biological activity is selected from the group consisting of: (i) upregulation of NF-&kgr;B; (ii) ability of the RANK polypeptide to bind a RANKL polypeptide comprising amino acids 69-317 of SEQ ID NO:13; (iii) ability of the RANK polypeptide to bind TRAF1, TRAP 2, TRAP 3, TRAP 5 or TRAP 6; (iv) upregulation of a reporter gene that is operably linked to a NF-&kgr;B-dependent promoter; and (v) in vitro clustering of CD34+ CD1a+ dendritic cells that have been contacted with a RANKL polypeptide comprising amino acids 69-317 of SEQ ID NO:13; and c) determining that the molecule is a RANK agonist if the biological activity is increased in the presence of the molecule or that the molecule is a RANK antagonist if the biological activity is decreased in the presence of the molecule.