Assay system to identify therapeutic agents
The invention is an assay designed to identify agents that modulate apoptosis. The invention provides an assay system and methods for screening for inhibitors of the Bcl-2 family of proteins. In various aspects the invention provides an assay system containing a liposome reagent and an immobilized BH3 domain peptide. In further aspects the invention provides an assay system containing mitochondria, an immobilized BH3 domain peptide and a mitochondrial binding agent, e.g. an anti-VDAC anti-body.
1. An assay system comprising:
a) an isolated mitochondria; and
b) a solid substrate comprising:
i) an immobilized BH3 domain peptide of less than 195 amino acids and
ii) an immobilized anti-VDAC antibody.
2. The assay system of claim 1 , wherein said mitochondria is isolated from a cancer cell.
3. The assay system of claim 1 , wherein said mitochondria is labeled with a potentiometric dye.
4. The assay system of claim 3 , wherein said potentiometric dye is JC-1 or dihydrorhodamine 123.
5. The assay system of claim 1 , wherein said BH3 domain peptide is an activating BH3 peptide.
6. The assay system of claim 1 , wherein said BH3 domain peptide is a sensitizing BH3 peptide.
7. The assay system of claim 1 , wherein said BH3 domain peptide comprises an amino acid sequence selected from SEQ ID NO: 1-7.
8. The assay system of claim 1 , wherein said solid substrate is a microarray or a microsphere.