Detection of BCL-2 family heterodimer complexes and use thereof
AssigneeThe United States of America, as represented by the Secretary, Department of Health and Human Services
InventorsApurva Kumar Srivastava, Dominic Esposito, Jeevan Prasaad Govindharajulu, Ralph Ewin Parchment, James Halpern Doroshow
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Abstract
Immunoassays to detect the presence of Bcl-2 family heterodimeric complexes are described. The immunoassays are designed to detect one or more of BIM-Bcl-2, BIM-Bcl-xL, BIM-Mcl-1 and BAX BAK heterodimers. The assays can be used, for example, to select a BH3 mimetic, or other drug targeting the apoptosis pathway, that is effective for treating a specific cancer, or to select a subject who is likely to respond to a particular BH3 mimetic.
Claims20
A method of detecting at least two different Bcl-2 family heterodimeric protein complexes in a biological sample, comprising:
- providing a cell lysate of a biological sample that comprises cells;
- and
- detecting the presence of the at least two different Bcl-2 family heterodimeric protein complexes in the cell lysate, wherein each heterodimeric protein complex comprises a first protein and a second protein, and wherein detecting each heterodimeric protein complex comprises:
- providing an antibody specific for the first protein bound to a solid support;
- providing a detection antibody specific for the second protein, wherein the detection antibody is covalently attached to a detectable label;
- contacting the cell lysate with the antibody-bound solid support and the detection antibody;
- and
- detecting the presence of the detection antibody, thereby detecting the Bcl-2 family heterodimeric complex in the biological sample,
- wherein the at least two different Bcl-2 family heterodimeric protein complexes comprise:
- BIM-Bcl-2 and BIM-Bcl-xL;
- BIM-Bcl-2 and BAK-BAK;
- BIM-Bcl-xL and BIM-Mcl-1;
- BIM-Bcl-xL and BAK-BAK;
- or
- BIM-Mcl-1 and BAX-BAK.
A method of detecting at least two different Bcl-2 family heterodimeric protein complexes in a biological sample that comprises cells, wherein the biological sample comprises a tissue sample, a biopsy sample, a fine-needle tumor aspirate, a bone marrow aspirate or a blood sample, comprising:
- providing a cell lysate of the biological sample;
- and
- detecting the presence of the at least two different Bcl-2 family heterodimeric protein complexes in the cell lysate, wherein each heterodimeric protein complex comprises a first protein and a second protein, and wherein detecting each heterodimeric protein complex comprises:
- providing an antibody specific for the first protein bound to a solid support;
- providing a detection antibody specific for the second protein;
- contacting the cell lysate with the antibody-bound solid support and the detection antibody;
- and
- detecting the presence of the detection antibody, thereby detecting the Bcl-2 family heterodimeric complex in the biological sample,
- wherein the at least two different Bcl-2 family heterodimeric protein complexes comprise:
- BIM-Bcl-2 and BIM-Bcl-xL;
- BIM-Bcl-2 and BAK-BAK;
- BIM-Bcl-xL and BIM-Mcl-1;
- BIM-Bcl-xL and BAK-BAK;
- or
- BIM-Mcl-1 and BAX-BAK.
Dependencies are inferred from the claim text.