Compositions and methods for capture of cellular targets of bioactive agents
View Patent ↗The present invention provides compositions and methods for capture and identification of the cellular targets of a bioactive agent. In particular, provided herein are bioactive agents tethered to capture ligand, cellular targets (optionally tagged with a reporter), capture proteins (optionally present as capture dimers), surfaces (e.g., displaying, capture ligands, capture proteins, or capture dimers), and methods of capturing and identifying the cellular targets of a bioactive agent therewith.
1. A kit comprising:
(a) a cell that expresses a fusion of a bioluminescent reporter and a cellular target of a small molecule bioactive agent, wherein the cellular target is located within a cell;
(b) the small molecule bioactive agent tethered by a carbamate linker to a chloroalkane capture ligand, wherein the small molecule bioactive agent tethered to a chloroalkane capture ligand is cell permeable, and wherein the small molecule bioactive agent is capable of non-covalently binding to the cellular target upon interaction thereof under intracellular conditions; and
(c) a solid surface displaying a capture protein, wherein the capture protein comprises a modified dehalogenase configured to covalently bind to the chloroalkane capture ligand upon interaction thereof.
2. The kit of claim 1 , wherein the bioluminescent reporter comprises a polypeptide with at least 70% sequence identity with SEQ ID NO.: 3.
3. The kit of claim 1 , wherein the capture protein comprises at least 70% sequence identity with SEQ ID NO.: 1.
4. The kit of claim 1 , wherein the solid surface is selected from the list consisting of: a well, a tube, a slide, a plate, a matrix, resin or a bead.
5. The kit of claim 4 , wherein the solid surface is magnetic or paramagnetic.