US 12,130,282 B2
Universal fluorescence assay of high affinity ligand transport
Phillip E. Klebba, Manhattan, KS (US); Somnath Chakravorty, Buffalo, NY (US); Yan Shipelskiy, Holland, PA (US); Salete M. Newton, Manhattan, KS (US); and Ashish Kumar, Manhattan, KS (US)
Assigned to Kansas State University Research Foundation, Manhattan, KS (US)
Appl. No. 17/253,412
Filed by Kansas State University Research Foundation, Manhattan, KS (US)
PCT Filed Jun. 18, 2019, PCT No. PCT/US2019/037760
§ 371(c)(1), (2) Date Dec. 17, 2020,
PCT Pub. No. WO2019/246118, PCT Pub. Date Dec. 26, 2019.
Claims priority of provisional application 62/686,464, filed on Jun. 18, 2018.
Prior Publication US 2021/0263014 A1, Aug. 26, 2021
Int. Cl. G01N 33/573 (2006.01); G01N 33/50 (2006.01); G01N 33/543 (2006.01)
CPC G01N 33/5008 (2013.01) [G01N 33/54366 (2013.01); G01N 2333/795 (2013.01)] 30 Claims
 
1. A universal bacterial assay for detecting and monitoring changes in concentration of a target analyte in solution, said assay comprising:
creating a reaction solution comprising a transport-deficient high affinity protein-based sensor and a test bacteria dispersed in an aqueous solution, wherein the sensor comprises a transport-deficient bacterial high affinity binding protein modified with a detectable label that generates a detectable signal;
adding a target analyte to said reaction solution, wherein said high affinity binding protein is specific for binding with said target analyte but transport-deficient, and wherein said target analyte is a metal or metallated complex;
exposing said reaction solution to an energy source to generate said detectable signal; and
detecting changes in the detectable signal in the reaction solution over time, wherein said changes correspond to interaction of said test bacteria with said target analyte.