Devices and methods for vertical flow-based detection of analytes
Provided herein are devices and methods for isolation and detection of an analyte from a sample. In some embodiments, provided herein are devices and methods of use thereof for flow-based isolation and detection of analytes in a liquid sample, such as a urine sample, that leverage both substantially vertical and substantially lateral flow paths within the device to enable sensitive and specific detection of analytes from large sample volumes.
1 . A device for detecting one or more analytes in a liquid sample, the device comprising a plurality of porous materials, wherein the device is configured such that a liquid sample flows substantially vertically through at least one of the plurality of porous materials and substantially laterally through at least one of the plurality of porous materials, wherein the plurality of porous materials comprise:
a) a filter system comprising one or more filter components;
b) at least one wicking component;
b) c) a porous membrane containing one or more capture moieties held within a defined capture region of the porous membrane; and
d) at least one absorbent pad,
wherein the device is configured such that the liquid sample flows through the filter system prior to contacting the at least one wicking component, travels substantially laterally in a first direction through the at least one wicking component and onto the porous membrane containing one or more capture moieties, flows through the porous membrane containing one or more capture moieties and into the at least one absorbent pad, and subsequently travels substantially laterally in a second direction through the absorbent pad, wherein the second direction is substantially opposite of the first direction.
2 . The device of claim 1 , further comprising a barrier between:
the at least one wicking component and the at least one absorbent pad, such that the sample does not enter into the absorbent pad prior to flowing through the porous membrane containing one or more capture moieties.
3 . The device of claim 1 , wherein the plurality of porous materials further comprise at least one sample pad, wherein the liquid sample is introduced into the device by adding the liquid sample to the at least one sample pad.
4 . The device of claim 3 , wherein the sample travels through the at least one sample pad and subsequently travels substantially vertically through the filter system.
5 . The device of claim 3 , wherein the sample travels through the at least one sample pad in both lateral and vertical flows paths.
6 . The device of claim 3 , wherein at least one sample pad comprises a reservoir which temporarily holds a volume of sample when added to the sample pad.
7 . The device of claim 3 , comprising at least two sample pads, wherein the at least two sample pads are stacked such that at least a portion of a first sample pad overlaps with at least a portion of a second sample pad.
8 . The device of claim 3 , wherein the filter system is housed on a lid of the device, and wherein a body of the device comprises at least one sample pad, the porous membrane containing one or more capture moieties held within a defined capture region of the porous membrane, the at least one wicking component, and the at least one absorbent pad.
9 . The device of claim 1 , further comprising one or more detection reagents added to or dried onto a component of the device.
10 . The device of claim 9 , wherein the one or more detection reagents are added to or dried onto a component of the device downstream of the filter system.
11 . The device of claim 9 , wherein the one or more detection reagents are added to or dried onto a reagent pad or a portion of the wicking component.
12 . The device of claim 1 , wherein at least one filter component comprises a high protein-binding membrane.
13 . The device of claim 12 , wherein the at least one filter component comprises a nylon membrane.
14 . The device of claim 1 , wherein the filter system comprises at least two filter components, wherein each filter component comprises pores and/or capture moieties for removal of contaminants.
15 . The device of claim 1 , further comprising one or more microfluidic channels, wherein at least one microfluidic channel transports the liquid sample to the capture region of the porous membrane.
16 . The device of claim 15 , further wherein at least one microfluidic channel transports the liquid sample to a control region of the porous membrane.
17 . The device of claim 15 , wherein the one or more microfluidic channels comprise a mechanism for air escape.
18 . The device of claim 1 , wherein the porous membrane comprising one or more capture moieties further contains one or more detection moieties.
19 . The device of claim 1 , further comprising at least one component to prevent the liquid sample from circumventing the filter system.
20 . A method of detecting one or more analytes in a liquid sample, the method comprising:
a. applying the liquid sample to the device of claim 1 , such that the analyte, if present in the sample, binds to the one or more capture moieties; and
b. detecting the one or more analytes, if present in the sample.
21 . The method of claim 20 , wherein one or more detection moieties are present within the device and/or are added to the device such that the one or more detection moieties binds to the analyte, thereby producing a detectable signal within the defined capture region of the porous membrane.
22 . The method of claim 21 , wherein the one or more detection moieties are contained within the porous membrane containing one or more capture moieties or the one or more detection moieties are added to the liquid sample prior to applying the liquid sample to the device.
23 . The method of claim 20 , wherein detecting the one or more analytes comprises assessing the detectable signal within the defined capture region.