Method and compositions for making ACSF and ACSF antagonists
AssigneeThe University of Melbourne
InventorsWilliam I. Wood, Thomas John Martin, Larry John Suva
Life of this patent
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Abstract
The complete amino acid and nucleotide sequence for adenylate cyclase stimulating factor is provided, thereby facilitating the synthesis of ACSF in recombinant cell culture. ACSF amino acid sequence variants and ACSF antibodies are provided which are useful in the treatment of humoral hypercalcemia of malignancy or in immunoassays for ACSF. In particular, antibodies capable of binding only the C-terminal domains of ACSF are useful in immunoassays for ACSF which avoid interference by parathyroid hormone. Also provided are novel polypeptides selected from the group of the ACSF basic peptide, the ACSF C-terminal peptide, or the ACSF domain containing both of the basic and C-terminal peptides.
Claims25
An isolated nucleic acid which encodes Adenylate Cyclase Stimulating Factor (ACSF) comprising the primary amino acid sequence of FIG. 2.
An isolated nucleic acid which encodes a polypeptide comprising about residues 1-83 of the primary amino acid sequence of FIG. 2.
An isolated nucleic acid which encodes a polypeptide comprising about residues 84-108 of the primary amino acid sequence of FIG. 2.
An isolated nucleic acid which encodes a polypeptide comprising about residues 109-141 of the primary amino acid sequence of FIG. 2.
An isolated nucleic acid which encodes an Adenylate Cyclase Stimulating Factor (ACSF) insertion variant polypeptide selected from the group consisting of [V.sub.2 GGS.sub.3 ]ACSF(1-141), [V.sub.2 SS.sub.3 ]ACSF(1-141), [V.sub.2 EKA.sub.3 ]ACSF(1-141), [G.sub.123 PPD.sub.124 ]ACSF(3-141), [S.sub.130 FYT.sub.131 ]ACSF(3-141), [R.sub.139 DYR.sub.140 ]ACSF(3-141), [V.sub.2 AGS.sub.3 ]ACSF(1-141), [T.sub.132 KKKS.sub.133 ]ACSF(1-141), [S.sub.3 EEE.sub.4 ]ACSF(134), [E.sub.4 IH.sub.5 ]ACSF(1-34), [E.sub.4 IH.sub.5 DQ.sub.6 ]ACSF(1-34) and [P.sub.44 DN.sub.45 ]ACSF(1-141).
An isolated nucleic acid which encodes an Adenylate Cyclase Stimulating Factor (ACSF) deletion variant polypeptide selected from the group consisting of ACSF(1-83), ACSF(1-34), ACSF(1-84, 109-141), ACSF(1-34, 40-141), ACSF(1-50, 60-141), ACSF(1-75, 84-141), ACSF(1-109), ACSF(3-34), ACSF(4-34), ACSF(5-34), ACSF(6-34) and ACSF(3-124).
An isolated nucleic acid which encodes an Adenylate Cyclase Stimulating Factor (ACSF) substitution variant polypeptide selected from the group consisting of PTH(7-34)ACSF(35-141) and PTH(3-34)ACSF(35-141), wherein PTH is parathyroid hormone.
An isolated nucleic acid which encodes an Adenylate Cyclase Stimulating Factor (ACSF) substitution variant polypeptide selected from the group consisting of [MY.sub.1 ]ACSF(1-141), [MP.sub.1 ]ACSF(1-141), [MG.sub.1 ]ACSF(1-141), [F.sub.2 ]ACSF(1-141), [Y.sub.2 ]ACSF(1-141), [H.sub.2 ]ACSF(1-141), [D.sub.3 ]ACSF(1-141), [Y.sub.3 ]ACSF(1-141), [H.sub.3 ]ACSF(1-141), [K.sub.20 ]ACSF(1-141), [E.sub.19 ]ACSF(1-141), [V.sub.21 ]ACSF(1-141), [D.sub.20 ]ACSF(1-141), [Y.sub.24 ]ACSF(1-141), [K.sub.25 ]ACSF(1-141), [E.sub.25 ]ACSF(1-141), [M.sub.31 ]ACSF(1-141), [Y.sub.34 ]ACSF(3-141), [I.sub.8, I.sub.18, Y.sub.34 ]ACSF(3-141), [I.sub.8, I.sub.18, Y.sub.34 ]ACSF(3-34), [D.sub.79 ]ACSF(1-141), [P.sub.98 ]ACSF(1-141), [P.sub.105 ]ACSF(1-141), [Y.sub.90 ]ACSF(1-141), [W.sub.89 ]ACSF(1-141), [H.sub.96 ]ACSF(1-141), [F.sub.110 ]ACSF(106-141), [Y.sub.117 ]ACSF(106-141), [D.sub.122 ]ACSF(106-141), [K.sub.125 ]ACSF(106-141), [Y.sub.132 ]ACSF(106-141), [A.sub.133 ]ACSF(106-141) and [D.sub.141 ]ACSF(106-141).
A method for making Adenylate Cyclase Stimulating Factor (ACSF) comprising:
- (a) isolating nucleic acid encoding a polypeptide comprising the primary amino acid sequence of FIG. 2;
- (b) inserting the nucleic acid into a replicable vector under the transcriptional control of a promoter;
- (c) transfecting the vector into a host cell which transcribes the nucleic acid under the control of the promoter;
- (d) culturing the cell to permit ACSF to accumulate in the cell culture and
- (e) recovering the ACSF from the cell culture.
Dependencies are inferred from the claim text.