US 2021/0263014 A1 · UNIVERSAL FLUORESCENCE ASSAY OF HIGH AFFINITY LIGAND TRANSPORT

Abstract

A universal bacterial assay and kit for detecting and monitoring changes in concentration of a target analyte in solution. The assay relies on an engineered high affinity protein-based sensor with a detectable label that has high specificity to a target analyte, typically a ligand or metallated complex, and can be used to spectroscopically monitor interaction of the target analyte with a test bacterial strain. The sensors can be in the form of living, transport-deficient bacterial cells expressing the engineered high affinity protein, or in the form of purified proteins themselves. The system can further be used to screen candidate compounds for interference with the binding and uptake of such analytes by bacterial cells, which is useful in investigation of new classes of antibiotics.

Claims30

Dependent claims

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 high affinity protein-based sensor and a test bacteria dispersed in an aqueous solution, wherein the sensor comprises a bacterial high affinity binding protein engineered with a detectable label that generates a detectable signal;
adding a target analyte to said reaction solution, wherein high affinity binding protein is specific for said target analyte, 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 strain with said target analyte.

2 more elements

Dependencies are inferred from the claim text.

Universal fluorescence assay of high affinity ligand transport

Life of this patent

Universal fluorescence assay of high affinity ligand transport

Life of this patent

Abstract

Claims30