University of California

Daniel Takashi Kamei

Timothy J Deming

Benjamin Ming Wu

Yin To Chiu

Garrett L Mosley

Zachary D Taylor

Eric P Holowka

Daniel William Bradbury

Maie St John

Ashley Elizabeth Kita

Shin Ting Sherine Frieda Cheung

Matthew Foosing Yee

Nguyen Khoi Le

Vincent K Wong

David Yuan Pereira

Joshua Anthony Keefe

Christina Caroline Pearce

Chloe Michelle Wu

Amir Adam Dailamy

Victor Z Sun

Disclosed herein are devices and methods that use aqueous two phase systems and lateral flow assays to detect target analytes in a sample. These devices and methods may be used to diagnose a disease or condition in a biological sample, such as blood or serum. In addition, these devices and methods may be used to detect allergens in a food samples or contaminants, such as environmental toxins, in water samples. Device and kit components may be conveniently assembled in a portable container and are amenable to actuation in most settings. The devices are simple to use, requiring a non-trained operator to simply add the sample to the device. Conveniently, the time it takes to detect the target analyte is very short. Thus, the devices and methods disclosed herein provide novel and useful means for point-of-care.

Methods and devices for integrating analyte extraction, concentration and detection

In various embodiments devices and methods for the detection and/or quantification of clinically relevant pathogens (e.g., bacteria, fungi, viruses, etc.) are provided. In certain embodiments the device comprises a lateral-flow assay that detects the bacterium at a concentration of less than about 6×10cells/mL, less than about 3×10cells/ml, less than about 1×10CFU/mL, or less than about 50 μg/mL. In certain embodiments the device comprises an aqueous two-phase system (ATPS) comprising a mixed phase solution that separates into a first phase solution and a second phase solution; and a lateral-flow assay (LFA). In certain embodiments the device comprises a flow-through system comprising a concentration component comprising an aqueous two-phase system (ATPS) comprising a mixed phase solution that separates into a first phase solution and a second phase solution; and a detection component disposed beneath said concentration component.

Methods and devices for analyte collection, extraction, concentration, and detection for clinical applications

Devices and methods are provided for the detection of CSF in a biological sample. In certain embodiments the device is a lateral flow device comprising: a porous substrate; a sample addition zone disposed on or in said porous substrate; a detection zone disposed on or in said porous substrate where said detection zone comprises at least a first test line (T) and a second test line (T) each test line comprising binding moieties that bind a complex formed between beta-trace protein (βTP) and an indicator attached to a βTP binding molecule; wherein said porous substrate defines a flow path through which a sample applied to the sample addition zone flows under capillary action away from said sample addition zone into said detection zone; and wherein said first test line (T) and said second test line (T) are is configured so that either no test line signal or just a signal at the first test line (T) is detectable when βTP concentration in a sample applied to said device is lower than the βTP level indicative of a CSF leak; and said second test line (T) is configured so that a signal is detectable at said second test line when βTP concentration in a sample applied to said device is greater than the βTP level indicative of a CSF leak.

Semi-quantitative lateral-flow immunoassay for the detection of CSF leaks

In various embodiments methods and devices are provided for the detection and/or quantification of an analyte. In certain embodiments a device is provided comprising an aqueous two-phase system (ATPS) comprising a mixed phase solution that separates into a first phase solution and a second phase where, in use, said first phase solution becomes a leading phase and said second phase solution becomes a lagging phase; a lateral-flow assay (LFA); and a probe and/or a development reagent, where in use, said probe associates with said first phase solution in said leading phase of said ATPS and/or said development reagent associates with said second phase solution in said lagging phase of said ATPS. In certain embodiments a 'one-pot' system of purifying and amplifying a nucleic acid is provided utilizing, e.g., an ATPS and isothermal amplification reagents.

Biomarker concentration and signal amplification for use in paper-based immunoassays and a single platform for extracting, concentrating, and amplifying DNA

It was discovered that hydrogel scaffolds can be used to induce phase separation as aqueous two-phase systems (ATPSs) pass through and/or rehydrate the scaffolds, allowing for concentration of target analyte(s) (e.g., biomolecule(s)) into a particular phase of the ATPS or into a leading front. Accordingly, in various embodiments methods and devices are provided that utilize aqueous two-phase systems and hydrogel scaffolds to improve the sensitivity of assays (e.g., of point-of-care assays) without sacrificing cost or ease of use.

Hydrogel platform for aqueous two-phase concentration of a target to enhance its detection

In various embodiments devices and methods for the detection and/or quantification of clinically relevant pathogens (e.g., bacteria, fungi, viruses, etc.) are provided. In certain embodiments the device comprises a lateral-flow assay that detects the bacterium at a concentration of less than about 6&#xd7;10cells/mL, less than about 3&#xd7;10cells/ml, less than about 1&#xd7;10CFU/mL, or less than about 50 &#x3bc;g/mL. In certain embodiments the device comprises an aqueous two-phase system (ATPS) comprising a mixed phase solution that separates into a first phase solution and a second phase solution; and a lateral-flow assay (LFA). In certain embodiments the device comprises a flow-through system comprising a concentration component comprising an aqueous two-phase system (ATPS) comprising a mixed phase solution that separates into a first phase solution and a second phase solution; and a detection component disposed beneath said concentration component.

Devices and methods are provided for the detection of CSF in a biological sample. In certain embodiments the device is a lateral flow device comprising: a porous substrate; a sample addition zone disposed on or in said porous substrate; a detection zone disposed on or in said porous substrate where said detection zone comprises at least a first test line (T) and a second test line (T) each test line comprising binding moieties that bind a complex formed between beta-trace protein (betaTP) and an indicator attached to a betaTP binding molecule; wherein said porous substrate defines a flow path through which a sample applied to the sample addition zone flows under capillary action away from said sample addition zone into said detection zone.

Growing community of inventors

Monterey Park, CA, United States of America

Daniel Takashi Kamei