Device for detecting misfolded proteins and methods of use thereof
Inventors
Davis, Wendy L. • Levenson, Daniel
Assignees
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Publication Number
US-11073516-B2
Publication Date
2021-07-27
Expiration Date
Abstract
The present invention relates to diagnostic devices as well as methods of using these devices for detecting proteins of interest associated with diseases or disorders in mammals. In particular, the proteins of interest may be misfolded proteins associated with certain misfolded-protein disorders in mammals including those mammals suspected of or at risk of having such disorders.
Core Innovation
The invention provides a diagnostic device and a method of detecting at least one protein in a biological sample of a mammal using a lateral-flow chromatographic diagnostic approach. A biological sample is applied to a sample receiving material and the at least one protein binds to a detection reagent present on or within the sample receiving material. The detection reagent is selected from an azo dye, Thioflavin T, or an analog of an azo dye.
The diagnostic device further includes a trap positioned in contact with the sample receiving material, where the trap separates detection reagent bound to the protein from unbound detection reagent. The trap permits flow of detection reagent bound to the protein through the trap while retaining, retarding, or binding the unbound detection reagent. The trap material is selected from nitrocellulose, cellulose, a glass fiber, a cotton fiber, a woven mesh, a nonwoven material, asymmetric polysulfone membrane, polyvinylidene fluoride, and polyethersulfone.
Downstream, a capillary bed is configured to contain the biological sample after the sample flows through the trap, with the sample receiving material, trap, and capillary bed configured to be in contact in sequence. The capillary bed displays detection reagent if the at least one protein is present in the biological sample, and detecting the presence of detection reagent on the capillary bed indicates presence of the protein in the biological sample.
The description addresses detection of misfolded proteins and protein aggregates, including supramolecular protein aggregates and congophilic proteins having beta-sheet structure, in biological samples. It also frames a clinical need for identifying protein-related conditions such as preeclampsia, including mild, severe, atypical preeclampsia, HELLP syndrome, and eclampsia, using a device suitable for mammalian samples such as pregnant mammal urine.
Claims Coverage
The document provides one independent claim. It defines a sequential lateral-flow style diagnostic device with three inventive features: a sample pad supporting a protein-binding detection reagent, a trap that removes unbound detection reagent while allowing bound detection reagent to pass, and a capillary bed that displays detection reagent such that presence of detection reagent indicates presence of the protein.
Protein-binding detection reagent on sample receiving material
The diagnostic device comprises a sample receiving material capable of receiving a biological sample, and a detection reagent present on or within the sample receiving material that binds at least one protein in the biological sample, wherein the detection reagent is selected from the group consisting of an azo dye, Thioflavin T and an analog of an azo dye.
Trap separating bound from unbound detection reagent
The diagnostic device comprises a trap in contact with the sample receiving material, wherein the trap separates detection reagent bound to at the at least one protein from unbound detection reagent, permits flow of detection reagent bound to the at least one protein through the trap, and retains, retards the flow of, or binds the unbound detection reagent.
Capillary bed displaying detection reagent to indicate protein presence
The diagnostic device comprises a capillary bed in contact with the trap, configured to contain the biological sample after the biological sample flows through the trap, wherein the sample receiving material, trap, and capillary bed are configured to be in contact in sequence, and wherein the capillary bed displays detection reagent if the at least one protein is present in the biological sample; detecting presence of the detection reagent on the capillary bed indicates presence of the at least one protein.
Across the independent claim, the coverage centers on a lateral-flow chromatographic sequence in which a detection reagent binds the target protein on a sample receiving material, a trap retains unbound detection reagent while allowing bound detection reagent to pass, and a capillary bed displays the detection reagent so that detection reagent presence on the capillary bed indicates protein presence.
Stated Advantages
Improved simplicity/stability versus prior paper-based assays.
Rapid readout (approximately 1–10 minutes).
Documented Applications
Detecting misfolded proteins and protein aggregates, including congophilic proteins having beta-sheet structure, in mammalian biological samples using a lateral-flow chromatographic diagnostic device.
Detecting preeclampsia in pregnant mammal urine, including mild preeclampsia, severe preeclampsia, atypical preeclampsia, HELLP syndrome, and eclampsia.
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