Compositions and methods for capture of cellular targets of bioactive agents

Patent metadata

Field	Value
Publication number	US-10168323-B2
Application number	US-201414204868-A
Country	US
Kind code	B2
Filing date	Mar 11, 2014
Priority date	Mar 15, 2013
Publication date	Jan 1, 2019
Grant date	Jan 1, 2019

Abstract

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.

First claim

The invention claimed is:

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.

Assignees

• Promega Corp

Inventors

• Hitko Carolyn W
• Hurst Robin
• Kirkland Thomas
• Klaubert Dieter
• Meisenheimer Poncho
• Nath Nidhi
• Ohana Rachel Friedman
• Otto Paul
• Urh Marjeta
• Uyeda Harry Tetsuo
• Wood Keith

Classifications

• G01N33/5005

  involving human or animal cells (immunoassay G01N33/56966; immunoassays of protozoa G01N33/56905; protozoa in screening assays C12Q1/025) · CPC title

• G01N33/543 Primary

  with an insoluble carrier for immobilising immunochemicals · CPC title