Detection of the degree of exposure to chemical warfare nerve agents and organophosphate pesticides with lateral flow assays
Inventors
Basinger, JR., G. William • Clement, Kristin H.
Assignees
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Publication Number
US-10288614-B2
Publication Date
2019-05-14
Expiration Date
Abstract
A sample analysis device used to detect a level of exposure of organophosphorus within a sample comprising a sample collection pad, at least one conjugate zone comprising an anti-analyte antibody that is conjugated with a reporter label, and a control antibody that is conjugated with a reporter label, a blocking and/or test zone comprising an immobilized nanoparticle or other molecule that captures the Organophosphate-bound analyte, a second blocking and/or test zone comprising an immobilized antibody that binds to the unbound analyte and an optional third blocking and/or control zone comprising an immobilized antibody that binds to the control molecule wherein, when the analyte is bound by the Organophosphate in the sample it will bind to the first test line, and if the analyte is “free’ from the Organophosphate it will bind to the second test line, and the control antibody will bind to the control line.
Core Innovation
The invention provides a baseline-free method of detecting a level of exposure to an organophosphorus compound by measuring both OP bound and unbound cholinesterase levels in a sample. The method includes collecting a sample and transferring the sample to a sample analysis device comprising a sample collection pad, a conjugate zone, and one or more blocking and/or test zones.
The device uses anti-analyte antibodies conjugated with a reporter label together with a control antibody conjugated with a reporter label. The anti-analyte antibody is selected from anti-acetylcholinesterase (AChE) antibody or anti-butylcholinesterase (BChE) antibody, and the zones distinguish OP-bound analyte from unbound analyte.
A first blocking and/or test zone includes an immobilized nanoparticle or other molecule that captures the Organophosphate-bound analyte, while a second blocking and/or test zone includes an immobilized antibody that binds to the unbound analyte. An optional third blocking and/or control zone includes an immobilized antibody that binds to a control molecule, so that bound analyte binds the first zone, free analyte binds the second zone, and the control antibody binds the third zone.
Detection is performed by detecting the presence or absence of the binding molecule in the first blocking and/or test zone and the remaining analyte in the second blocking and/or test zone to culminate reporting a ratio of bound- and free-analyte. The method then determines the level of OP exposure for an individual based on OP binding state rather than using a predetermined baseline measurement.
Claims Coverage
The patent provides two independent claims: a method claim and a sample analysis device claim. Across these claims, the inventive features center on measuring OP-bound versus OP-free cholinesterase (AChE/BChE) using a multi-zone lateral flow/dipstick style device and reporting a bound-to-free ratio for individual exposure level determination.
OP bound and unbound cholinesterase measurement for exposure level
Detecting a level of exposure to an organophosphorus compound via measurement of organophosphate (OP) bound and unbound cholinesterase levels in a sample, comprising collecting a sample and transferring it to a sample analysis device that distinguishes bound versus free analyte.
Conjugate zone with reporter-labeled anti-analyte and control antibodies
A conjugate zone comprising an anti-analyte antibody conjugated with a reporter label and a control antibody conjugated with a reporter label, where the anti-analyte antibody is selected from anti-acetylcholinesterase (AChE) antibody or anti-butylcholinesterase (BChE) antibody.
First and second test zones for OP-bound versus unbound analyte binding
A first blocking and/or test zone comprising an immobilized nanoparticle or other molecule that captures the Organophosphate-bound analyte, and a second blocking and/or test zone comprising an immobilized antibody that binds to the unbound analyte, with bound analyte binding the first zone and free analyte binding the second zone.
Optional control zone for control molecule binding
An optional third blocking and/or control zone comprising an immobilized antibody that binds to a control molecule, wherein the control antibody binds to the third blocking and/or control zone.
Ratio reporting of bound and free analyte and individual exposure determination
Detecting presence or absence of the binding molecule in the first blocking and/or test zone and the remaining analyte in the second blocking and/or test zone to culminate reporting a ratio of bound- and free-analyte, and determining the level of OP exposure for an individual.
Sample analysis device with multi-zone binding and ratio reporting
A sample analysis device comprising a sample collection pad, at least one conjugate zone with reporter-labeled anti-analyte and control antibodies, a first blocking and/or test zone capturing the Organophosphate-bound analyte, a second blocking and/or test zone binding the unbound analyte, and an optional third blocking and/or control zone binding a control molecule, to detect first and second zone binding to report a ratio of bound- and free-analyte and determine OP exposure level for an individual.
Across the independent claims, the invention centers on a multi-zone sample analysis device that uses reporter-labeled anti-cholinesterase antibodies to distinguish OP-bound versus unbound AChE/BChE, captures OP-bound analyte in a first zone using an immobilized nanoparticle or other molecule, captures free analyte in a second zone using an immobilized antibody, optionally includes a control zone, and reports a ratio of bound to free analyte to determine OP exposure level for an individual.
Stated Advantages
Not requiring a predetermined baseline measurement, by reporting a ratio of bound- and free-analyte.
Documented Applications
Point-of-care detection of exposure to organophosphorus compounds by determining an individual’s level of OP exposure using a baseline-free cholinesterase binding assay.
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