US 12,123,053 B2
Molecular glue screening assays and methods for practicing same
Ksenya Cohen Katsenelson, La Jolla, CA (US); and Daniel Kelly Treiber, San Diego, CA (US)
Assigned to Eurofins DiscoverX Corporation, San Diego, CA (US)
Filed by Eurofins DiscoverX Corporation, San Diego, CA (US)
Filed on Jan. 25, 2022, as Appl. No. 17/584,315.
Claims priority of provisional application 63/143,255, filed on Jan. 29, 2021.
Prior Publication US 2022/0243255 A1, Aug. 4, 2022
Int. Cl. C12Q 1/686 (2018.01); C12N 9/00 (2006.01)
CPC C12Q 1/686 (2013.01) [C12N 9/93 (2013.01)] 19 Claims
 
1. A method of identifying a test compound as molecular glue, wherein the test compound stabilizes interaction between two proteins not previously known to interact, said method comprising the steps of:
(a) contacting a cell or cell lysate comprising a fusion protein with the test compound, wherein the fusion protein comprises a target protein of interest and a nucleic acid oligomer tag;
(b) incubating the fusion protein with the test compound in the presence of an immobilized ligand protein, wherein the ligand protein is immobilized on a solid support;
(c) removing any unbound target protein of interest from the cell or cell lysate; and
(d) detecting an amount of the fusion protein that binds to the immobilized ligand protein by detecting the nucleic acid oligomer tag bound to the fusion protein; wherein an increase in the amount of the fusion protein of step (d) detected in the presence of the said test compound relative to the amount of the fusion protein detected in the absence of the said test compound indicates that the said test compound modulates the interaction between two proteins not previously known to interact.