US10302650B2 – May 28, 2019 – High-throughput fluorescence polarization microorganism screening assay

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Inventors :

Yao Zong Ng; Pedro Alexis Baldera Aguayo; Virginia W. Cornish; Ehud Herbst

Owner :

THE TRUSTEES OF COLUMBIA UNIVERSITY IN THE CITY OF

Application Number :

US16008777

Document Number :

US10302650B2

Priority Date :

June 14, 2018

Filing Date :

June 14, 2018

Date of Grant/ Publication :

May 28, 2019

Class :

; G01N33 / 58; G01N21 / 64; C12N15 / 10; G01N33 / 569; C40B40 / 02; C12Q1 / 68; G01N33 / 542

Abstract

The present disclosure provides a high-throughput fluorescence polarization microorganism screening assay for a metabolite produced by a microorganism. The metabolite is detected using competition with a fluorescent reporter for a receptor that binds both the metabolite and the reporter. The present disclosure further provides materials used in such an assay, a kit for such an assay, an isolated microorganism identified using such an assay, and culture conditions identified using such an assay.

Claim(s)

1. A method of high-throughput fluorescence polarization microorganism screening comprising: culturing a plurality of microorganism strains that produce a metabolite in a plurality of separate compartments to produce a plurality of samples; adding a receptor and fluorescent reporter to the plurality of samples obtained from the plurality of compartments; detecting fluorescence polarization of the reporter in the plurality of samples in a multi-compartment fluorescence polarization assay device, wherein fluorescence polarization of the reporter correlates inversely with a concentration of the metabolite in any given sample because the metabolite competes with the fluorescent reporter to bind to the receptor and binding of the fluorescent reporter to the receptor increases fluorescence polarization; calculating concentration of the metabolite in at least a portion of the plurality of samples; and designating at least one of the plurality of microorganism strains as high-producing or low-producing based on the calculated concentration of metabolite for at least one sample.;
43. A method of high-throughput fluorescence polarization microorganism screening comprising: culturing a microorganism strain that produces a metabolite under plurality of different culture conditions in a plurality of separate compartments to produce a plurality of samples; adding a receptor and fluorescent reporter to the plurality of samples obtained from the plurality of compartments; detecting fluorescence polarization of the reporter in the plurality of samples in a multi-compartment fluorescence polarization assay device, wherein fluorescence polarization of the reporter correlates inversely with a concentration of the metabolite in any given sample because the metabolite competes with the fluorescent reporter to bind to the receptor and binding of the fluorescent reporter to the receptor increases fluorescence polarization; calculating concentration of the metabolite in at least a portion of the plurality of samples; and designating at least one of the plurality of culture conditions as high-producing or low-producing based on the calculated concentration of metabolite for at least one sample.

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