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US 6,319,675 B1Granted
Methods for detecting and/or identifying agents which bind and/or modulate function of “bonzo” chemokine receptor
AssigneeMillennium Pharmaceuticals, Inc.
InventorsMichael J. Briskin, Kristine E. Murphy, Alyson M. Wilbanks, Lijun Wu
Life of this patent
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Abstract
The invention relates to a method of detecting and/or identifying agents (molecules, compounds) which can bind to Bonzo and inhibit the binding of a ligand and/or modulate a function of Bonzo.
Claims58
A method of detecting and/or identifying an agent which binds to Bonzo or a SExCkine-binding variant of Bonzo comprising combining:
- a) SExCkine or a Bonzo-binding variant of SExCkine,
- b) a test agent, and
- c) a composition comprising Bonzo or a SExCkine-binding variant of Bonzo under conditions suitable for binding of said SExCkine or Bonzo-binding variant to said Bonzo or SExCkine-binding variant;
- and
- detecting or measuring the formation of a complex between said SExCkine or Bonzo-binding variant and said Bonzo or SExCkine-binding variant, wherein a decrease in the formation of said complex relative to a suitable control indicates that said test agent binds to said Bonzo or SExCkine-binding variant,
- wherein said SExCkine or Bonzo-binding variant is selected from the group consisting of (i) SEQ ID NO:4, (ii) SEQ ID NO:6, (iii) a Bonzo-binding fragment of SEQ ID NO:4, (iv) a Bonzo-binding fragment of SEQ ID NO:6 and (v) a Bonzo-binding polypeptide having 90% amino acid sequence similarity over the full length of said Bonzo-binding polypeptide to any one or (i)-(iv);
- and
- wherein said Bonzo or SExCkine-binding variant is selected from the group consisting of (1) SEQ ID NO:2, (2) a SExCkine-binding fragment of SEQ ID NO:2 and (3) a SExCkine-binding polypeptide having 90% amino acid sequence similarity over the full length of said SExCkine-binding polypeptide to (1) or (2).
5 more elements
A method of detecting and/or identifying an antagonist of Bonzo comprising combining;
- a) a cell expressing Bonzo or a SExCkine-binding variant of Bonzo;
- b) SExCkine or a Bonzo-binding variant of SExCkine;
- and
- c) an agent to be tested, under conditions suitable for detecting a SExCkine- or Bonzo-binding variant-induced response;
- and
- determining the ability of the test agent to inhibit said response, wherein inhibition of said response by the agent indicates that the agent is an antagonist,
- wherein said Bonzo or SExCkine-binding variant is selected from the group consisting of (i) SEQ ID NO:2, (ii) a SExCkine-binding fragment of SEQ ID NO:2 and (iii) a SExCkine-binding polypeptide having 90% amino acid sequence similarity over the full length of said SExCkine-binding polypeptide to (i) or (ii);
- and
- wherein said SExCkine or Bonzo-binding variant is selected from the group consisting of (1) SEQ ID NO:4, (2) SEQ ID NO:6, (3) a Bonzo-binding fragment of SEQ ID NO:4, (4) a Bonzo-bindinig fragment of SEQ ID NO:6 and (5) a Bonzo-binding polypeptide having 90% amino acid sequence similarity over the full length of said Bonzo-binding polypeptide to any one of (1)-(4).
6 more elements
A method of detecting and/or identifying an agent which binds to Bonzo or a SExCkine-binding variant of Bonzo comprising combining:
- a) a fusion protein comprising SExCkine or a Bonzo-binding variant of SExCkine,
- b) a test agent, and
- c) a composition comprising Bonzo or a SExCkine-binding variant of Bonzo, under conditions suitable for binding of said SExCkine or Bonzo-binding variant to said Bonzo or SExCkines-binding variant;
- and
- detecting or measuring the formation of a complex between said SExCkine or Bonzo-binding variant and said Bonzo or SExCkine-binding variant, wherein a decrease in the formation of said complex relative to a suitable control indicates that said test agent binds to said Bonzo or SExCkine-binding variant,
- wherein said SExCkine or Bonzo-binding variant is selected from the group consisting of (i) SEQ ID NO:4, (ii) SEQ ID NO:6, (iii) a Bonzo-binding fragment of SEQ ID NO:4, (iv) a Bonzo-binding fragment of SEQ TD NO:6 and (v) a Bonzo-binding polypeptide having 90% amino acid sequence similarity over the full length of said Bonzo-binding polypeptide to any one of (i)-(iv);
- and
- wherein said Bonzo or SExCkine-binding variant is selected from the group consisting of (1) SEQ ID NO:2, (2) a SExCkine-binding fragment of SEQ ID NO:2 and (3) a SExCkine-binding polypeptide having 90% amino acid sequenee similarity over the full length of said SExCkine-binding polyeptide to (1) or (2).
5 more elements
A method of detecting and/or identifying an agent which binds to Bonzo or a SExCkine-binding variant of Bonzo comprising combining:
- a) a first fusion protein comprising SExCkine or a Bonzo-binding variant of SExCkine,
- b) a test agent, and
- c) a composition comprising a second fusion protein, said second fusion protein comprising Bonzo or a SExCkine-binding variant of Bonzo, under conditions suitable for binding of said SExCkine or Bonzo-binding variant to said Bonzo or SExCkine-binding variant;
- and
- detecting or measuring the formation of a complex between said SExCkine or Bonzo-binding variant and said Bonzo or SExCkine-binding variant wherein a decrease in the formation of said complex relative to a suitable control indicates that said test agent binds to said Bonzo or SExCkine-binding variant,
- wherein said SExCkine or Bonzo-binding variant is selected from the group consisting of (i) SEQ ID NO:4, (ii) SEQ ID NO:6, (iii) a Bonzo-binding fragment of SEQ ID NO:4, (iv) a Bonzo-binding fragment of SEQ ID NO:6 and (v) a Bonzo-binding polypeptide having 90% amino acid sequence similarity over the full length of said Bonzo-binding polypeptide to any one of (i)-(iv);
- and
- wherein said Bonzo or SExCkine-binding variant is selected from the group consisting of (1) SEQ ID NO:2, (2) a SExCkine-binding fragment of SEQ ID NO:2 and (3) a SExCkine-binding polypeptide having 90% amino acid sequence similarity over the full length of said SExCkine-binding polypeptide to (1) or (2).
5 more elements
A method of detecting and/or identifying an agent which binds to Bonzo or a SExCkine-binding variant of Bonzo comprising combining:
- a) SFxCkine or a Bonzo-binding variant of SExCkine,
- b) a test agent, and
- c) a composition comprising a fusion protein that comprises Bonzo or a SExCkine binding variant of Bonzo under conditions suitable for binding of said SExCkine or Bonzo-binding variant to said Bonzo or SExCkine-binding variant;
- and
- detecting or measuring the formation of a complex between said SExCkine or Bonzo-binding variant and said Bonzo or SExCkine-binding variant, wherein a decrease in the formation of said complex relative to a suitable control indicates that said test agent binds to said Bonzo or SixCkine-binding variant,
- wherein said SExCkine or Bonzo-binding variant is selected from the group consisting of (i) SEQ ID NO:4, (ii) SEQ ID NO:6, (iii) a Bonzo-binding fragment of SEQ ID NO:4, (iv) a Bonzo-binding fragment of SEQ ID NO:6 and (v) a Bonzo-binding polypeptide having 90% amino acid sequence similarity over the full length of said Bonzo-binding polypeptide to any one of (i)-(iv);
- and
- wherein said Bonzo or SExCkine-binding variant is selected from the group consisting of (1) SEQ ID NO:2, (2) a SExCkine-binding fragment of SEQ ID NO:2 and (3) a SExCkine-binding polypeptide having 90% amino acid sequence similarity over the full length of said SExCkine-binding polypeptide to (1) or (2).
5 more elements
A method of detecting and/or identifying an antagonist of Bonzo comprising combining:
- a) a cell expressing a fusion protein comprising Bonzo or a SExCkine-binding variant of Bonzo;
- b) SExCkine or a Bonzo-binding variant of SExCkine, and
- c) an agent to be tested, under conditions suitable for detecting a SExCkine- or Bonzo-binding variant-induced response;
- and
- determining the ability of the test agent to inhibit said response, wherein inhibition of said response by the agent indicates that the agent is an antagonist,
- wherein said Bonzo or SExCkine-binding variant is selected from the group consisting of (i) SEQ ID NO:2, (ii) a SExCkine-binding fragment of SEQ ID NO:2 and (iii) a SExCkine-binding polypeptide having 90% amino acid sequence similarity over the full length of said SExCkine-binding polypeptide to (i) or (ii);
- and
- wherein said SExCkine or Bonzo-binding variant is selected from the group consisting of (1) SEQ ID NO:4, (2) SEQ ID NO:6, (3) a Bonzo-binding fragment of SEQ ID NO:4, (4) a Bonzo-binding fragment of SEQ ID NO:6 and (5) a Bonzo-binding polypeptide having 90% amino acid sequence similarity over the full length of said Bonzo-binding polypeptide to any one of (1)-(4).
5 more elements
A method of detecting and/or identifying an antagonist of Bonzo comprising combining:
- a) a cell expressing Bonzo or a SExCkine-binding variant of Bonzo;
- b) a fusion protein comprising SExCkine or a Bonzo-binding variant of SExCkine, and
- c) an agent to be tested, under conditions suitable for detecting a SExCkine- or Bonzo-binding variant-induced response;
- and
- determining the ability of the test agent to inhibit said response, wherein inhibition of said response by the agent indicates that the agent is an antagonist,
- wherein said Bonzo or SExCkine-binding variant is selected from the group consisting of (i) SEQ ID NO:2, (ii) a SExCkine-binding fragment of SEQ ID NO:2 and (iii) a SExCkine-binding polypeptide having 90% amino acid sequence similarity over the full length of said SExCkine-binding polypeptide to (i) or (ii);
- and
- wherein said SExCkine or Bonzo-binding variant is selected from the group consisting of (1) SEQ ID NO:4, (2) SEQ ID NO:6, (3) a Bonzo-binding fragment of SEQ ID NO:4, (4) a Bonzo-binding fragment of SEQ ID NO:6 and (5) a Bonzo-binding polypeptide having 90% amino acid sequence similarity over the full length of said Bonzo-binding polypeptide to any one of (1)-(4).
5 more elements
A method of detecting and/or identifying an antagonist or Bonzo comprising combining:
- a) a cell expressing a first fusion protein comprising Bonzo or a SExCkine-binding variant of Bonzo;
- b) a second fusion protein comprising SExCkine or a Bonzo-binding variant of SExCkine, and
- c) an agent to be tested, under conditions suitable for detecting a SExCkine- or Bonzo-binding variant-induced response;
- and
- determining the ability of the test agent to inhibit said response, wherein inhibition of said response by the agent indicates that the agent is an antagonist, and
- wherein said Bonzo or SExCkine-binding variant is selected front the group consisting of (i) SEQ ID NO:2, (ii) a SExCkine-binding fragment of SEQ ID NO:2 and (iii) a SExCkine-binding polypeptide having 90% amino acid sequence similarity over the full length of said SExCkine-binding polypeptide to (i) or (ii);
- and
- wherein said SExCkine or Bonzo-binding variant is selected from the group consisting of (1) SEQ ID NO:4, (2) SEQ ID NO:6, (3) a Bonzo-binding fragment of SEQ ID NO:4, (4) a Bonzo-bindinig fragment of SEQ ID NO:6 and (5) a Bonzo-binding polypeptide having 90% amino acid sequence similarity over the full length of said Bonzo-binding polypeptide to anyone of (1)-(4).
5 more elements
A method of detecting and/or identifying an antagonist of Bonzo comprising combining:
- a) a cell expressing a first fusion protein comprising Bonzo or a SExCkine-binding variant of Bonzo;
- b) a second fusion protein comprising a soluble SExCkine or a soluble Bonzo-binding variant of SExCkine, and
- c) an agent to be tested, under conditions suitable for detecting a SExCkine- or Bonzo-binding variant-induced response;
- and
- determining the ability of the test agent to inhibit said response, wherein inhibition of said response by the agent indicates that the agent is an antagonist, and
- wherein said Bonzo or SExCkine-binding variant is selected from the group consisting of (i) SEQ ID NO:2, (ii) a SExCkine-binding fragment of SEQ ID NO:2 and (iii) a SExCkine-binding polypeptide having 90% amino acid sequence similarity over the full length of said SExCkine-binding polypeptide to (i) or (ii);
- and
- wherein said SExCkine or Bonzo-binding variant is selected from the group consisting of (1) SEQ ID NO:4, (2) SEQ ID NO:6, (3) a Bonzo-binding fragment of SEQ ID NO:4, (4) a Bonzo-binding fragment of SEQ ID NO:6 and (5) a Bonzo-binding polypeptide having 90% amino acid sequence similarity over the full length of said Bonzo-binding polypeptide to any one of (1)-(4).
5 more elements
Dependencies are inferred from the claim text.