High throughput compatible assay for receptor-TRAF interactions
AssigneeBoehringer Ingelheim Pharmaceuticals, Inc.
InventorsMarilyn R. Kehry, Steven S. Pullen, James J. Crute
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Abstract
Disclosed is a high throughput compatible assay that is useful for the identification of specific antagonists of TRAF-receptor interactions. The modular flexibility of the assay makes it possible to introduce simple modifications in order to measure the interaction of any TNF receptor cytoplasmic domain (or TRAF-binding protein) with any of the six TRAF proteins, TRAF1, TRAF2, TRAF3, TRAF4, TRAF5 and TRAF6.
Claims7
A quantitative assay for measuring the ability of an antagonist or agonist to effect binding of a TRAF protein or fragment thereof to a TRAF interacting receptor having a cytosolic binding domain or fragment thereof, the TRAF protein or fragment thereof comprising SEQ ID NO:
- 1 residues 1-240 or SEQ ID NO:2 residues 1-225, the assay comprising:
- providing the receptor having a cytosolic binding domain or fragment thereof, wherein the receptor is bound to a multi-well plate, the multi-well plate allowing formation of a multimeric receptor and allowing detection of a signal;
- providing the TRAF protein or fragment thereof possessing a terminal tag, the tag comprising one or more amino acids and allowing binding a signal-generating antibody or fragment thereof;
- providing the antagonist or agonist;
- combining the receptor bound to the well, the TRAF protein and the antagonist or agonist;
- detecting the signal-generating antibody and measuring the inhibition or enhancement of the antagonist or agonist.
Dependencies are inferred from the claim text.