Cell-based assay for the detection of toxic analytes
View Patent ↗Cell-based reagents and methods of using the reagents for detecting analytes are provided. The adsorbing of cells with signal-generating metabolic activity to solid supports has been found to improve the sensitivity of known cell based assays. Signal-generating cells adsorbed to a solid support are introduced to a test agent, and the measured decrease in metabolic signal provides a measure of the toxicity of the test agent.
1. A method of detecting a potentially toxic analyte comprising:
(a) measuring a signal generated by preparing a test suspension comprising the potentially toxic analyte to be detected with a cellular reagent in the form of a preparation of eukaryotic or prokaryotic cells having signal-generating metabolic activity, wherein said cellular reagent is adsorbed onto a particulate solid-phase carrier having a particle size of 500 μm or less;
(b) measuring a control signal generated by preparing a control suspension comprising the cellular reagent without the potentially toxic analyte; and
(c) comparing the signals generated in (a) and (b), wherein a decrease in the signal in (a) as compared to (b) indicates detection of the toxic analyte.
2. The method of claim 1 , wherein said cellular reagent is subjected to a stabilization step prior to being used in said method.
3. The method of claim 2 , wherein said stabilization step comprises freeze-drying a mixture of said cellular reagent and a particulate carrier.
4. The method of claim 1 wherein said signal-generating metabolic activity is bioluminescence and the particulate solid-phase carrier is inorganic.
5. The method of claim 4 wherein said preparation of prokaryotic or eukaryotic cells are cells which have been genetically modified to exhibit bioluminescence.
6. The method of claim 5 wherein said cells have been genetically modified to contain a nucleic acid encoding luciferase.
7. The method of claim 1 wherein said preparation of prokaryotic cells are bacteria.
8. The method of claim 7 wherein said bacteria are Vibrio fischeri.
9. The method of claim 1 wherein the particulate solid phase carrier is particulate silica.
10. The method of claim 1 , wherein the particulate solid phase carrier comprises an organic or inorganic particulate.
11. The method of claim 1 wherein the particulate solid phase carrier is selected from the group consisting of polymers, alumina and cellulose.
12. A method of preparing to assay a potentially toxic analyte, said method comprising immobilizing or adsorbing onto a particulate solid-phase carrier a preparation of eukaryotic or prokaryotic cells having signal-generating metabolic activity wherein the solid-phase carrier has a particle size of 500 μm or less.
13. The method of claim 12 , wherein the eukaryotic or prokaryotic cells having signal-generating metabolic activity are subjected to a stabilization step prior to being used in the method.
14. The method of claim 13 , wherein the stabilization step comprises freeze-drying a mixture of the eukaryotic or prokaryotic cells having signal-generating metabolic activity and the particulate carrier.