Intracellular complexes as biomarkers
First Claim
1. A method of determining disease status of a patient suffering from a disease characterized by aberrant expression of one or more intracellular complexes, the method comprising the steps of:
- measuring directly in a patient sample an amount of each of one or more intracellular complexes;
comparing each such amount to its corresponding amount in a reference sample; and
correlating differences in the amounts from the patient sample and the respective corresponding amounts from the reference sample to the disease status the patient.
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Abstract
The invention provides a method for determining a disease status of a patient by measuring expression levels of selected intracellular complexes. In one aspect of the invention, the activation status of apoptotic pathways in a patient sample is determined by measuring relative amounts of protein-protein complexes that are characteristic of the apoptotic pathways. In particular, the invention provides a method of determining the activation status of the mitochondrial apoptotic pathway by simultaneously measuring relative amounts of complexes between 14-3-3 proteins and BAD proteins on the one hand and complexes of Bcl-2 proteins and BAD proteins on the other hand. Preferably, methods of the invention are implemented by using sets of binding compounds having releasable molecular tags that are specific for multiple components of one or more complexes formed within apoptotic pathways. After binding, molecular tags are released and separated from the assay mixture for analysis.
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Citations
23 Claims
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1. A method of determining disease status of a patient suffering from a disease characterized by aberrant expression of one or more intracellular complexes, the method comprising the steps of:
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measuring directly in a patient sample an amount of each of one or more intracellular complexes;
comparing each such amount to its corresponding amount in a reference sample; and
correlating differences in the amounts from the patient sample and the respective corresponding amounts from the reference sample to the disease status the patient. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16)
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- 17. A method of determining an apoptotic status of cells in a sample, the method comprising the step of simultaneously measuring amounts of at least one complex selected from the group consisting of a complex comprising a Bcl-2 protein and a BH3-only protein and a complex comprising a 14-3-3 protein and a BAD protein.
- 21. A method of determining a status of a cancer in a patient, the method comprising the step of simultaneously measuring in a sample from the patient amounts of at least one intracellular complex selected from the group consisting of a first complex comprising a Bcl-2 protein and a BH3-only protein and a second complex comprising a 14-3-3 protein and a BAD protein.
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23. A method of determining an apoptotic status of cells in a sample, the method comprising the step of simultaneously measuring amounts of at least one complex selected from the group consisting of a complex comprising an NF-kB protein and an I-kB protein and amounts of free NF-KB proteins and free I-kB proteins.
Specification