US 11,899,021 B2
Luciferase-based thermal shift assays
Melanie Dart, Madison, WI (US); Lance P. Encell, Fitchburg, WI (US); Thomas Kirkland, Atascadero, CA (US); Thomas Machleidt, Madison, WI (US); Matthew Robers, Madison, WI (US); Brock F. Binkowski, Sauk City, WI (US); Keith Wood, Mt. Horeb, WI (US); and Ce Shi, San Luis Obispo, CA (US)
Assigned to Promega Corporation, Madison, WI (US)
Filed by Promega Corporation, Madison, WI (US)
Filed on Jan. 4, 2023, as Appl. No. 18/150,018.
Application 16/787,950 is a division of application No. 15/017,271, filed on Feb. 5, 2016, granted, now 10,571,471, issued on Feb. 25, 2020.
Application 18/150,018 is a continuation of application No. 17/105,024, filed on Nov. 25, 2020, granted, now 11,567,083.
Application 17/105,024 is a continuation of application No. 16/787,950, filed on Feb. 11, 2020, granted, now 10,928,400, issued on Feb. 23, 2021.
Claims priority of provisional application 62/112,518, filed on Feb. 5, 2015.
Prior Publication US 2023/0341410 A1, Oct. 26, 2023
This patent is subject to a terminal disclaimer.
Int. Cl. C12Q 1/66 (2006.01); C12Q 1/68 (2018.01); G01N 21/64 (2006.01); G01N 33/53 (2006.01); G01N 33/58 (2006.01); G01N 33/68 (2006.01); G01N 33/569 (2006.01)
CPC G01N 33/6845 (2013.01) [C12Y 113/12007 (2013.01); G01N 21/6428 (2013.01); G01N 33/56916 (2013.01); G01N 33/581 (2013.01); G01N 33/582 (2013.01); G01N 2021/6432 (2013.01); G01N 2021/6439 (2013.01); G01N 2333/90241 (2013.01); G01N 2500/04 (2013.01); G01N 2500/10 (2013.01)] 11 Claims
 
1. A method to detect an interaction between a ligand and a target protein, comprising the steps of:
(a) incubating (i) a fusion of a target protein and a peptide tag comprising 70% or greater sequence identity with VSGWRLFKKIS (SEQ ID NO: 5) with (ii) a complementary polypeptide comprising 70% or greater sequence identity with SEQ ID NO: 6, wherein the peptide tag and the complementary polypeptide are capable of forming a bioluminescent complex:
(i) in the presence of the ligand to produce a test sample, and
(ii) in the absence of the ligand, to produce a control sample;
(b) treating said test and control samples under conditions that cause the target protein to unfold partially or completely unfold;
(c) measuring signal from the bioluminescent complex in said test and control samples; and
(d) comparing the measurement made in step (c) between the test and control samples, wherein alteration of the signal from said bioluminescent complex in the test sample compared to the control sample indicates the presence of the interaction between the ligand and the target protein.