Assays and methods for evaluating multimeric complexes
AssigneeWyeth
InventorsXiaoke Yang, Lori J. Fitz, Julie M. Lee, Jonathan Brooks, Stanley F. Wolf
Life of this patent
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Abstract
Assays, e.g., homogenous assays, and methods for identifying, quantifying and/or monitoring the formation and/or stability of a multimeric complex, e.g., a ternary complex are disclosed. The methods and assays of the invention can be used to identify and/or evaluate agents (e.g., proteins, peptides, antibody molecules, and small and large molecules) that interfere with and/or inhibit the formation of a multimeric complex (e.g., a ternary complex) or that disrupt a previously formed complex.
Claims24
A method for evaluating the formation or stability of a ternary complex, comprising:
- providing a sample that comprises at least three binding members under conditions that allow the formation of the ternary complex to occur;
- detecting, quantifying or monitoring a change in the level of the ternary complex using a homogeneous proximity-based detection method,
- wherein the formation or stability of the ternary complex is evaluated over a specified time interval, or in the presence of a test agent relative to a reference sample.
A homogenous assay for evaluating the formation or stability of a ternary complex, comprising:
- providing a sample that comprises at least three binding members under conditions that allow the formation of the ternary complex to occur;
- detecting, quantifying or monitoring a change in the level of the ternary complex using a proximity-based detection method,
- wherein the formation or stability of the ternary complex is evaluated over a specified time interval, or in the presence of a test agent relative to a reference sample.
A method of identifying an agent that modulates the formation or stability of a ternary complex, comprising:
- contacting a sample that comprises at least three binding members with a test agent under conditions that allow the formation of the ternary complex to occur;
- detecting, quantifying or monitoring the presence of the complex in the sample contacted with the test agent relative to a reference sample using a homogeneous proximity-based detection method,
- wherein a change in the level of the complex in the presence of the test agent, relative to the level of the complex in the reference sample, indicates that said test agent affects the formation or stability of said complex.
An assay for identifying an agent that modulates the formation or stability of a ternary complex, comprising:
- contacting a sample that comprises at least three binding members with a test agent under conditions that allow the formation of the ternary complex to occur;
- detecting, quantifying or monitoring the presence of the complex in the sample contacted with the test agent relative to a reference sample using a homogeneous proximity-based detection method,
- wherein a change in the level of the complex in the presence of the test agent, relative to the level of the complex in the reference sample, indicates that said test agent affects the formation or stability of said complex.
A method for identifying one or more members within a multimeric complex, comprising:
- detectably identifying a library of candidate binding members;
- detectably identifying at least one known member of the complex;
- contacting said identified library with said identified at least one member of the complex, under conditions that allow an interaction to occur,
- wherein the interaction of the library member with the at least one member of the complex results in a detectable signal.
Dependencies are inferred from the claim text.