Device, system and method for concentration of targets in samples using osmotic and capillary forces

Abstract

A device, a system, and a method for using the device and system is provided. The device, system, and method are useful for reducing volume of a sample while retaining an analyte of interest in the sample, thereby increasing concentration of the analyte in the reduced volume sample. The reduced volume sample may be analyzed for presence of the analyte of interest. The reduced volume sample may be analyzed for determining concentration of the analyte of interest.

Description

[0001] DEVICE, SYSTEM AND METHOD FOR CONCENTRATION OF TARGETS IN SAMPLES

[0002] USING OSMOTIC AND CAPILLARY FORCES

[0003] CROSS-REFERENCE TO RELATED APPLICATIONS

[0004] This application claims priority benefit to the filing date of U.S. Provisional Patent Application Serial No. 63 / 529,619, filed on July 28, 2023, the disclosure of which application is herein incorporated by reference in its entirety.

[0005] INTRODUCTION

[0006] A sample can be evaluated to determine presence of an analyte of interest. Determining presence of the analyte can be valuable in assessing environmental samples as well as biological samples. Fluidic samples may contain an analyte at low concentrations which may make the qualitative and / or quantitative analysis of the analyte inaccurate. Accurate detection of an analyte and / or quantitation of an analyte is important for detecting toxins, viruses, bacteria, and the like in environmental samples. Similarly, presence of an analyte in a biological fluid from a subject can inform on the health of the subject from which the sample is obtained. For example, presence of a virus or bacteria in the mucus or urine of a patient can be indicative of whether the patient has a viral or bacterial illness. Diagnostic assays, such as, lateral flow assays (LFAs) are used to determine whether an analyte of interest is present in a fluid sample. However, LFAs may not be sufficiently sensitive to detect an analyte of interest in a fluidic sample due to low concentration of the analyte. Thus, it is important to increase concentration of the analyte in the fluidic sample.

[0007] As such, concentration and / or enrichment of targets in samples is one of the key methods used to increase the sensitivity and limit of detection of a test / assay. Conventional methods employed for these processes include filtration and resuspension, positive and negative isolation using affinity / binding to particles, centrifugation among others. These methods either require active forces or driving mechanisms (centrifugation among others) or require multi-step protocols. For example, urine samples can be processed in multiple steps where assay chemistry / reagents including particles are typically isolated in different steps by the user. This leads to handling of the samples in an unsuitable manner with multiple points in the process that can lead to sample, target and / or reagent losses, exposure of the operator to the biohazard urine sample or harmful reagents and as a result, loss of efficiency in the sample preparation process. This also directly lowers assay quality for the intended diagnostic purpose of the sample.

[0008] Passive concentrators (that do not require active forces like centrifuges) based on semi-permeable osmotic membranes are available in the market typically for protein concentration (Minicon, Sartorius, BJP products among others). These methods typically use a membrane to allow passive osmosis of the sample solution through to an absorbent on the other side while selectively allowing for target concentration. However, these methods are prone to losses of target molecules to the surface of the membrane, or within the devices especially since localization of the target is not accomplished actively in these methods. Additionally, depending on the type of membrane, the losses can be even more pronounced for certain molecules to certain membranes. These concerns are even more pronounced in a decentralized setting with an untrained user where time is of essence.

[0009] Concentration of liquids like urine can be carried out using active and passive methods. Passive methods could include concentration using a semi-permeable Osmotic membrane that allows for selective passage of molecules through it based on their molecular weight, namely forward osmosis. Protein concentrators typically use this process to concentrate aqueous solutions / samples to enrich them with the target of concern. However, for rapid processing of samples (e.g., short time, large starting sample volume and high degree of concentration) passive methods can fall short of the required characteristics. FIG. 1A depicts concentration of a target molecule under standard forward osmotic conditions at the membrane surface. Samples can be concentrated by the passive flow; however, the localization of the target is limited due to the relatively large membrane surface area which is required for shorter processing times.

[0010] A key factor to reduce processing time is the ability to increase the contact surface of the sample with the membrane. However, this has an undesired effect where the target molecule is spread over the large surface area of the membrane. Therefore, there is a clear and unmet need to improve processing of a sample to increase analyte concentration.

[0011] SUMMARY

[0012] The present disclosure provides devices, systems, and methods that utilizes two types of forces to concentrate a sample and localize the target analyte to a relatively small area, thereby increasing recovery of the concentrated analyte. The concentrated sample may be analyzed for presence of the analyte of interest. The concentrated sample may be analyzed for determining amount of the analyte of interest.

[0013] Aspects of the invention include devices for reducing volume of a sample while retaining an analyte of interest in the sample is disclosed. Devices of interest include a chamber comprising side walls defining an opening; a mesh comprising a plurality of capillary channels disposed over the opening; a semi-permeable membrane disposed over the mesh and comprising pores permeable to water and substantially impermeable to analyte; and a hypertonic medium in contact with the semi-permeable membrane, wherein the device is configured for generating (a) a first concentration force driven by osmotic flow of water from a sample present in the chamber to the hypertonic medium across the mesh and the semi-permeable membrane, thereby reducing the volume of the sample, and (b) a second concentration force driven by a mesh-induced capillary action, wherein the second concentration force is orthogonal to the first concentration force, thereby moving solutes present in the sample towards a center of the chamber.

[0014] In some embodiments, devices of interest comprise an outlet positioned at a substantially central region of the device opposite the opening. In other embodiments, the outlet is flu idically connected to an extraction channel, wherein the extraction channel extends from the outlet to a sample extraction region accessible from the exterior of the device. In some embodiments, devices of interest comprise an inlet for entrance of the sample into the chamber, wherein the inlet is positioned adjacent the opening.

[0015] In some embodiments, devices of interest further comprise a lid in a fitting configuration with respect to the opening of the chamber. In other embodiments, devices of interest further comprise a sorbent retainer comprising the hypertonic medium. In still other embodiments, devices of interest further comprise a sorbent backing disposed over the sorbent retainer and under the lid. In yet other embodiments, devices of interest further comprise a sample. The sample may include a biological sample, an environmental sample, or a synthetic sample.

[0016] Aspects of the invention include systems for reducing volume of a sample while retaining an analyte of interest in the sample. In some embodiments, systems of interest comprise the device of the present disclosure and one or more of a sample analysis cartridge, a pipette, and instructions for reducing volume of a sample. The instructions may also include methods for analysis of the sample. Aspects of the invention include methods for reducing volume of a sample while retaining an analyte of interest in the sample. In some embodiments, methods of interest comprise processing the sample in the device of the present disclosure or the system of the present disclosure.

[0017] BRIEF DESCRIPTION OF THE FIGURES

[0018] FIG. 1 A depicts concentration of a target molecule at the membrane surface under standard forward osmotic conditions.

[0019] FIG. 1B depicts a method of the present disclosure that combines osmotic flowbased concentration with a localization process that allows for accumulation of the target molecule in a specific area.

[0020] FIG. 2A depicts an exemplary device 200 according to one embodiment of the present disclosure.

[0021] FIG. 2B depicts a cross-section of device 200 FIG. 2A and an enlarged view of the cross-section.

[0022] FIG. 3A depicts an exemplary device 300 according to one embodiment of the present disclosure.

[0023] FIG. 3B illustrates a cross-section of device 300 shown in FIG. 3A.

[0024] FIG. 3C illustrates an exploded view of device 300 shown in FIG. 3A.

[0025] FIG. 4 depicts an exemplary system 400 according to one embodiment comprising a test device 401 , a urine collection vessel 402, a test cartridge 403, and a pipette 404.

[0026] FIG. 5 depicts an exemplary process for reducing volume of a sample while retaining an analyte of interest in the sample by using a system depicted in FIG. 4.

[0027] FIG. 6 depicts an exemplary device 600 comprising a urine collection vessel 601.

[0028] FIG. 7 depicts an exemplary process for reducing volume of a sample while retaining an analyte of interest in the sample by use of a test device as provided herein and a urine collection vessel depicted in FIG. 6 according to one embodiment of the present disclosure.

[0029] FIG. 8A depicts an exemplary vacutainer urine collection vessel.

[0030] FIG. 8B depicts an exemplary test device which integrates with a test cartridge such as lateral flow test (LFT) card according to one embodiment of the present disclosure. FIG. 9 depicts an exemplary process for reducing volume of a sample while retaining an analyte of interest in the sample by use of a standard vacutainer urine collection vessel depicted in FIG. 8A and a test device depicted in FIG. 8B according to one embodiment of the present disclosure.

[0031] FIG. 10A depicts an exemplary test device and a sponge stick sampler. FIG. 10B depicts a sponge stick sampler inserted horizontally in a test device with an open lid with dashed lines for hidden outlines. FIG. 10C depicts a test device a sponge stick sampler inserted horizontally in a test device with a closed lid with dashed lines for hidden outlines.

[0032] FIG. 11 depicts an exemplary process for reducing volume of a sample while retaining an analyte of interest in the sample by use of a test device and a sponge stick sampler depicted in FIG. 10A according to one embodiment of the present disclosure.

[0033] FIG. 12A depicts an exemplary urine collection vessel.

[0034] FIG. 12B depicts an exemplary test device according to one embodiment of the present disclosure.

[0035] FIG. 13 depicts an exemplary process for reducing volume of a sample while retaining an analyte of interest in the sample by use of a urine collection vessel depicted in FIG. 12A and a test device depicted in FIG. 12B according to one embodiment of the present disclosure.

[0036] FIG. 14A depicts an exemplary urine collection pot with septum seal lid and funnel.

[0037] FIG. 14B depicts an exemplary test device according to one embodiment of the present disclosure.

[0038] FIG. 15 depicts an exemplary process for reducing volume of a sample while retaining an analyte of interest in the sample by use of a standard urine collection pot depicted in FIG. 14A and a test device depicted in FIG. 14B according to one embodiment of the present disclosure.

[0039] DETAILED DESCRIPTION

[0040] Aspects of the present disclosure include a device, a system, and a method for using the device and system. The device, system, and method are useful for reducing volume of a sample while retaining an analyte of interest in the sample. The reduced volume sample may be analyzed for presence of the analyte of interest. The reduced volume sample may be analyzed for determining concentration of the analyte of interest. Before the present devices, systems, and methods are described in greater detail, it is to be understood that the present disclosure is not limited to particular embodiments described, as such may, of course, vary. It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments only, and is not intended to be limiting.

[0041] Where a range of values is provided, it is understood that each intervening value, to the tenth of the unit of the lower limit unless the context clearly dictates otherwise, between the upper and lower limit of that range and any other stated or intervening value in that stated range, is encompassed within the present systems, devices and methods. The upper and lower limits of these smaller ranges may independently be included in the smaller ranges and are also encompassed within the systems, devices and methods, subject to any specifically excluded limit in the stated range. Where the stated range includes one or both of the limits, ranges excluding either or both of those included limits are also included in the systems, devices and methods.

[0042] Certain ranges are presented herein with numerical values being preceded by the term “about.” The term “about” is used herein to provide literal support for the exact number that it precedes, as well as a number that is near to or approximately the number that the term precedes. In determining whether a number is near to or approximately a specifically recited number, the near or approximating un-recited number may be a number which, in the context in which it is presented, provides the substantial equivalent of the specifically recited number.

[0043] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs. Although any methods and materials similar or equivalent to those described herein can also be used in the practice or testing of the present systems, devices and methods, representative illustrative systems, devices and methods are now described.

[0044] The present disclosure may be understood more readily by reference to the following detailed description of desired embodiments and the examples included therein. In the following specification and the claims which follow, reference will be made to a number of terms which shall be defined to have the following meanings.

[0045] The term “comprising” is used herein as requiring the presence of the named component and allowing the presence of other components. The term “comprising” should be construed to include the term “consisting essentially of” and “consisting of.” The “consisting essentially of” allows the presence of the named component(s), along with other component which do not change the function / structure of the named component(s). The “consisting of” allows the presence of the named component(s), along with any adhesives or other bonding means for attaching the listed component(s).

[0046] Numerical values should be understood to include numerical values which are the same when reduced to the same number of significant figures and numerical values which differ from the stated value by less than the experimental error of conventional measurement technique of the type described in the present application to determine the value.

[0047] All ranges disclosed herein are inclusive of the recited endpoint and independently combinable (for example, the range of “from 2 ml to 10 ml” is inclusive of the endpoints, 2 ml and 10 ml, and all the intermediate values). The endpoints of the ranges and any values disclosed herein are not limited to the precise range or value; they are sufficiently imprecise to include values approximating these ranges and / or values.

[0048] The modifier “about” used in connection with a quantity is inclusive of the stated value and has the meaning dictated by the context. When used in the context of a range, the modifier “about” should also be considered as disclosing the range defined by the absolute values of the two endpoints. For example, the range of from about “2 to about 10” also discloses the range “from 2 to 10.” The term “about” may refer to plus or minus 10% of the indicated number. For example, “about 10%” may indicate a range of 9% to 11 %, and “about 1” may mean from 0.9-1.1.

[0049] It should be noted that many of the terms used herein are relative terms. For example, the terms “top” and “bottom” are relative to each other in location and refer to surfaces where the top is always higher than the bottom relative to an absolute reference, i.e. , the surface of the earth. The terms “inlet” and “outlet” are relative to a fluid flowing through them with respect to a given structure, e.g., a fluid flows through the inlet into the structure and flows out of the structure through the outlet. The terms “upwards” and “downwards” are also relative to an absolute reference; upwards is always against the gravity of the earth while downwards is always towards the gravity of the earth.

[0050] The term “parallel” should be construed in its lay sense of two surfaces that maintain a generally constant distance between them, and not in the strict mathematical sense that such surfaces will never intersect when extended to infinity.

[0051] All publications and patents cited in this specification are herein incorporated by reference as if each individual publication or patent were specifically and individually indicated to be incorporated by reference and are incorporated herein by reference to disclose and describe the methods and / or materials in connection with which the publications are cited. The citation of any publication is for its disclosure prior to the filing date and should not be construed as an admission that the present invention is not entitled to antedate such publication by virtue of prior invention. Further, the dates of publication provided may be different from the actual publication dates which may need to be independently confirmed.

[0052] It is noted that, as used herein and in the appended claims, the singular forms “a”, “an”, and “the” include plural referents unless the context clearly dictates otherwise. It is further noted that the claims may be drafted to exclude any optional element. As such, this statement is intended to serve as antecedent basis for use of such exclusive terminology as “solely,” “only” and the like in connection with the recitation of claim elements or use of a “negative” limitation.

[0053] As will be apparent to those of skill in the art upon reading this disclosure, each of the individual embodiments described and illustrated herein has discrete components and features which may be readily separated from or combined with the features of any of the other several embodiments without departing from the scope or spirit of the present device, systems and methods. Any recited method can be carried out in the order of events recited or in any other order which is logically possible.

[0054] DEVICES FOR REDUCING SAMPLE VOLUME AND CONCENTRATING AN ANALYTE

[0055] As summarized above, aspects of the present disclosure include a device for reducing volume of a sample while retaining an analyte of interest in the sample, thereby increasing the concentration of the analyte relative to the concentration in the initial sample, is provided. In certain aspects, the device may include a chamber comprising side walls defining an opening. A mesh comprising a plurality of capillary channels may be disposed over the opening. A semi-permeable membrane may be disposed over the mesh. The semi-permeable membrane may include pores that are permeable to water and substantially impermeable to the analyte. A hypertonic medium may be disposed in the device such that it is in contact with the semi-permeable membrane. The device may be configured for generating (a) a first concentration force driven by osmotic flow of water from a sample present in the chamber to the hypertonic medium, across the mesh and the semi-permeable membrane, thereby reducing the volume of the sample, and (b) a second concentration force driven by a mesh-induced capillary action, wherein the second concentration force is orthogonal to the first concentration force, thereby moving solutes present in the sample towards a center of the chamber.

[0056] Sample Chamber

[0057] Devices of interest include a chamber comprising side walls defining an opening. In some embodiments, the chamber may be in shape of cylinder, sphere, ellipsoid, lentiform, hemisphere, ovoid, claviform, conical frustum, cuboid, cube, rectangular prism, triangular prism, pentagonal prism, hexagonal prism, nonagonal prism, octagonal prism, or truncated pyramid. In some embodiments, the cross-section of the chamber is in shape of circle, rectangle, square, oval, rhombus, pentagon, hexagon, nonagon, octagon, or polygon. In certain embodiments, the chamber may be substantially cylindrical in shape, e.g., a right angle cylinder, and may include a curved side wall extending between two opposite bases that are substantially parallel to each other and wherein an opening is formed in one of the bases.

[0058] In some embodiments, the cross-section of the chamber is in shape of circle and the diameter of chamber is, in some instances, about 5 cm or more, 10 cm or more, 15 cm or more, 20 cm or more, 30 cm or more, 40 cm or more, 40 cm or more, 50 cm or more, 60 cm or more, 70 cm or more, 80 cm or more, 90 cm or more, or 100 cm or more. For example, the diameter of the chamber is, for example, in the range of 5 cm to 500 cm, 5 cm to 400 cm, 5 cm to 300 cm, 5 cm to 200 cm, 5 cm to 150 cm, 5 cm to 100 cm, 5 cm to 90 cm, 5 cm to 80 cm, 5 cm to 70 cm, 5 cm to 60 cm, 5 cm to 50 cm, 5 cm to 40 cm, 5 cm to 30 cm, 5 cm to 25 cm, 5 cm to 20 cm, 5 cm to 10 cm, 7 cm to 20 cm, 8 cm to 20 cm, 10 cm to 20 cm, or 10 cm to 15 cm, etc.. The diameter of the chamber varies depending on several variables such as amount of a sample, a chamber volume, and a chamber height.

[0059] In some embodiments, the cross-section of the chamber is in shape of non-circle and width of the chamber across the largest cross-section is, in some instances, about 2 cm or more, about 3 cm or more, about 4 cm or more, about 5 cm or more, 10 cm or more, 15 cm or more, 20 cm or more, 30 cm or more, 40 cm or more, 40 cm or more, 50 cm or more, 60 cm or more, 70 cm or more, 80 cm or more, 90 cm or more, or 100 cm or more. For example, the width and / or length of the chamber is, for example, in the range of 5 cm to 500 cm, 5 cm to 400 cm, 5 cm to 300 cm, 5 cm to 200 cm, 5 cm to 150 cm, 5 cm to 100 cm, 5 cm to 90 cm, 5 cm to 80 cm, 5 cm to 70 cm, 5 cm to 60 cm, 5 cm to 50 cm, 5 cm to 40 cm, 5 cm to 30 cm, 5 cm to 25 cm, 5 cm to 20 cm, 5 cm to 10 cm, 7 cm to 20 cm, 8 cm to 20 cm, 10 cm to 20 cm, or 10 cm to 15 cm. The width / length of the chamber varies depending on several variables such as amount of a sample, a chamber volume, and a chamber height.

[0060] In some embodiments, the height of the chamber is, in some instances, about 1 cm or more, 2 cm or more, 3 cm or more, 4 cm or more, 5 cm or more, 10 cm or more, 20 cm or more, 30 cm or more, 50 cm or more, 100 cm or more. For example, the height of the chamber is, for example, in the range of 1 cm to 100 cm, 1 cm to 50 cm, 1 cm to 35 cm, 1 cm to 30 cm, 1 cm to 25 cm, 1 cm to 20 cm, 1 cm to 15 cm, 2 cm to 15 cm, 3 cm to 15 cm, 4 cm to 15 cm, 5 cm to 15 cm, 1 cm to 13 cm, 1 cm to 10 cm, 1 cm to 8 cm, 1 cm to 5 cm, 2 cm to 10 cm, 2 cm to 8 cm, 2 cm to 6 cm, 4 cm to 6 cm, etc. The height of the chamber varies depending on several variables such as amount of a sample, a chamber volume, and a chamber width.

[0061] In some embodiments, the diameter of the chamber is longer than the height of the chamber. For example, the diameter of the chamber is 1 .2 times or more, 1 .5 times or more, 1.8 times or more, 2.0 times or more, 2.3 times or more, 2.5 times or more, 3 times or more, 3.5 times or more, 3.8 times or more, 4 times or more, 4.5 times or more, 5 times or more, 5.5 times or more, 6 times or more, 6.5 times or more, 7 times or more, 7.5 times or more, 8 times or more, 8.5 times or more, 9 times or more, 9.5 times or more, 10 times or more longer than the height of the chamber. In other embodiments, the width / length of the chamber is longer than the height of the chamber. In some embodiments, the width and / or length of the chamber is longer than the height of the chamber. For example, the width and / or length of the chamber is 1 .2 times or more, 1 .5 times or more, 1 .8 times or more, 2.0 times or more, 2.3 times or more, 2.5 times or more, 3 times or more, 3.5 times or more, 3.8 times or more, 4 times or more, 4.5 times or more, 5 times or more, 5.5 times or more, 6 times or more, 6.5 times or more, 7 times or more, 7.5 times or more, 8 times or more, 8.5 times or more, 9 times or more, 9.5 times or more, 10 times or more longer than the height of the chamber.

[0062] In some embodiments, volume of the chamber is, in some instances, 5 ml or more, 8 ml or more, about 10 ml or more, 15 ml or more, 20 ml or more, 25 ml or more, 30 ml or more, 35 ml or more, 40 ml or more, 45 ml or more, 50 ml or more or 100 ml or more. In other embodiments, the volume of the chamber is, in some instances, in the range of 5 ml to 500 mL, 5 ml to 400 mL, 5 ml to 300 mL, 5 ml to 200 mL, 5 ml to 100 mL, 1 ml to 500 mL, 1 ml to 400 mL, 1 ml to 300 mL, 1 ml to 200 mL, 1 ml to 100 mL, 5 ml to 80 mL, 5 ml to 60 mL, 5 ml to 50 mL, 5 ml to 40 mL, 5 ml to 30 mL, 5 ml to 20 mL, 5 ml to 15 mL, 5 ml to 13 mL, or 1 ml to 10 ml. The volume of the chamber varies depending on several variables such as amount of a sample.

[0063] In certain embodiments, the chamber may be substantially cylindrical in shape and may include a curved side wall extending between two opposite bases that are substantially parallel to each other and wherein an opening is formed in one of the bases.

[0064] In some embodiments, the opening of the chamber is located on the top end of the chamber. In other embodiments, the opening is located on the top end of the chamber and extends to the side walls. In some embodiments, a mesh comprising a plurality of capillary channels is disposed over the opposite base of the opening of the chamber. In other embodiments, a semi-permeable membrane is disposed over the mesh such that it is in contact with the mesh. In still other embodiments, a hypertonic medium is disposed in the device such that it is in contact with the semi-permeable membrane.

[0065] In some embodiments, the opening of the chamber is located on the bottom end of the chamber. In other embodiments, the opening is located on the bottom end of the chamber and extends to the side walls. In some embodiments, a mesh comprising a plurality of capillary channels is disposed over the opposite base of the opening of the chamber. In other embodiments, a semi-permeable membrane is disposed over the mesh such that it is in contact with the mesh. In still other embodiments, a hypertonic medium is disposed in the device such that it is in contact with the semi-permeable membrane.

[0066] In some embodiments, the chamber is made of high-density materials such as brass, glass, acrylic, hard steel stainless steel, mild steel, titanium, copper, aluminum or combination thereof. In other embodiments, the chamber is made of lightweight materials, such as plastic.

[0067] Sample Outlet

[0068] Devices of interest include a base region of the sample chamber comprising an outlet. In some embodiments, the outlet is positioned at a substantially central region of the device opposite the opening of the sample chamber. In other embodiments, the outlet comprises a closed configuration and an open configuration. In certain embodiments, the closed configuration of the outlet comprises an air seal. In certain embodiments, the closed configuration comprises a pierceable seal. In certain embodiments, the closed configuration of the outlet comprises a diaphragm. In some embodiments, the outlet is flu idically connected to the extraction channel, wherein the extraction channel extends from the outlet to a sample extraction region accessible from the exterior of the device. In some embodiments, the extraction channel is positioned in the center of the base. A concentrated sample fluid can flow out through the extraction channel towards a sample extraction region accessible from the exterior of the device. In some embodiments, the sample extraction region is positioned on the exterior surface of the device.

[0069] When the outlet is in the closed configuration, air pressure exists in the extraction channel such that the sample fluid cannot flow through the extraction channel. When a sample extraction region which is covered by a pierceable seal is punctured by a sample collection device, the sample fluid can flow out through the extraction channel and the sample fluid is collected in the sample extraction region.

[0070] In some embodiments, the base of the chamber comprises one outlet. In other embodiments, the base of the chamber comprises two or more outlets. In certain cases, each outlet may be fluidically connected to each extraction channel respectively and the extraction channels are connected to one sample extraction region accessible from the exterior of the device. In certain cases, each outlet may be fluidically connected to one extraction channel and the extraction channel is connected to one sample extraction region accessible from the exterior of the device.

[0071] Sample Inlet

[0072] Devices of interest may further comprise an inlet for entrance of the sample into the chamber. In some embodiments, the inlet is the opening of the sample chamber. In some embodiments, the inlet is separate from the sample chamber opening. In some embodiments, the inlet is positioned adjacent the opening of the chamber. In some embodiments, the device of the present disclosure may comprise a lid in a fitting configuration with respect to the opening of the chamber. In certain cases, the inlet is positioned at the center of the lid. In certain cases, the inlet is positioned at a peripheral region of the lid. The inlet is fluidically connected to the sample chamber in the device. In some embodiments, a channel fluidically connects the inlet to the sample chamber.

[0073] In some embodiments, devices of interest may comprise a plurality of inlets for entrance of the sample into the chamber. In certain cases, a plurality of inlets is positioned at peripheral regions of the lid and fluidically connected to the chamber. In certain cases, a plurality of inlets is positioned at the center of the lid and fluidically connected to the chamber.

[0074] In certain embodiments, the inlet is configured to connect with an interfacing unit of a sample vessel to receive a sample without exposure to air. For example, the inlet is comprises a piercing member, e.g., a spike. The piercing member may include a sample entrance path to allow flow of sample from the punctured sample vessel to the sample chamber of the device. The sample vessel may comprise pierceable region on the bottom or top. In certain embodiments, the inlet comprises a piercing member extending from the inlet and configured for piercing a sample collection vessel pressed against the piercing member and providing the sample comprises pressing a bottom end or a top end of the sample collection vessel against the piercing member.

[0075] Mesh Component

[0076] Devices of interest include a mesh for providing a plurality of capillary channels. In some embodiments, the mesh is disposed over the opening of the chamber. In other embodiments, the mesh is in contact with sample present in the chamber and has sufficient porosity to allow the sample or a phase thereof (e.g. , the liquid phase) to contact a semipermeable membrane disposed over the mesh.

[0077] In some embodiments, the mesh extends to the side walls of the chamber. In certain embodiments, the mesh is substantially planar. In some embodiments, the shape of the substantially planar mesh matches the shape of cross section of the chamber, such as, circle, rectangle, square, oval, rhombus, pentagon, hexagon, nonagon, octagon, or polygon.

[0078] In some embodiments, the mesh is in shape of circle, the diameter of mesh is, in some instances, about 5 cm or more, 10 cm or more, 15 cm or more, 20 cm or more, 30 cm or more, 40 cm or more, 40 cm or more, 50 cm or more, 60 cm or more, 70 cm or more, 80 cm or more, 90 cm or more, or 100 cm or more. For example, the diameter of the mesh is, for example, in the range of 5 cm to 500 cm, 5 cm to 400 cm, 5 cm to 300 cm, 5 cm to 200 cm, 5 cm to 150 cm, 5 cm to 100 cm, 5 cm to 90 cm, 5 cm to 80 cm, 5 cm to 70 cm, 5 cm to 60 cm, 5 cm to 50 cm, 5 cm to 40 cm, 5 cm to 30 cm, 5 cm to 25 cm, 5 cm to 20 cm, 7 cm to 20 cm, 8 cm to 20 cm, 10 cm to 20 cm, or 10 cm to 15 cm. The diameter of the mesh varies depending on several variables such as amount of a sample, a chamber diameter, and the like. In other embodiments, mesh is not in shape of a circle and the width across the longest cross-section of the mesh is, in some instances, about 5 cm or more, 10 cm or more, 15 cm or more, 20 cm or more, 30 cm or more, 40 cm or more, 40 cm or more, 50 cm or more, 60 cm or more, 70 cm or more, 80 cm or more, 90 cm or more, or 100 cm or more. The width / length of the mesh varies depending on several variables such as amount of a sample, a chamber width / length, a chamber volume and the like.

[0079] In some embodiments, the mesh has a thickness in the range of about 50 pm to about 200 pm such as about 50 pm to about 190 pm, about 50 pm to about 180 pm, about 50 pm to about 170 pm, about 50 pm to about 160 pm, about 50 pm to about 150 pm, about 60 pm to about 200 pm, about 70 pm to about 200 pm, about 80 pm to about 200 pm, about 90 pm to about 200 pm, about 100 pm to about 200 pm, about 100 pm to about 150 pm, about 100 pm to about 140 pm, about 100 pm to about 130 pm, about 100 pm to about 125 pm. The thickness of the mesh varies depending on several variables such as amount of a sample, a chamber volume, and a chamber size.

[0080] In some embodiments, the mesh is made of polypropylene, polyester, nylon, spandex, woven, copper or combination thereof. In other embodiments, the mesh is a synthetic polymer. In other embodiments, the mesh forms a film including capillary channels for capillary action. In some embodiments, the mesh is a micromesh.

[0081] In some embodiments, the mesh comprises a plurality of capillary channels such that a mesh-induced capillary action generates a second concentration force. In some embodiments, the second concentration force is orthogonal to the first concentration force, thereby moving solutes present in the sample towards a center of the chamber.

[0082] In other embodiments, the mesh comprises a plurality of pores. In some embodiments, the pore size is less than 1 mm. The pore size varies depending on the size of solutes present in a sample.

[0083] Semi-permeable membrane

[0084] Devices of interest include semi-permeable membrane. In some embodiments, the semi-permeable membrane includes pores that are permeable to water and substantially impermeable to the analyte of interest.

[0085] A semi-permeable membrane may be disposed over the mesh. The semi- permeable membrane may include pores that are permeable to water and substantially impermeable to the analyte. In some embodiments, the semi-permeable membrane comprises a plurality of pores. In certain cases, the pores extending through the semi-permeable membrane may be large enough such that a fluid in the fluid sample (e.g., water) can pass through the pores, but may be small enough such that target analytes and / or solutes in the solution cannot pass through the pores. The pore size varies depending on the size of solutes present in a sample. For example, the pore size is in the range of 1 nm to 500 nm, 1 nm to 450 nm, 1 nm to 400 nm, 1 nm to 300 nm, 1 nm to 350 nm, 1 nm to 300 nm, 1 nm to 200 nm, 1 nm to 250 nm, 1 nm to 100 nm, 1 nm to 90 nm, 1 nm to 80 nm, 1 nm to 70 nm, 1 nm to 60 nm, 1 nm to 50 nm, 1 nm to 40 nm, 1 nm to 30 nm, 1 nm to 20 nm, or 1 nm to 10 nm.

[0086] In some embodiments, the semi-permeable membrane has a thickness in the range of about 5 pm to about 100 pm such as about 5 pm to about 90 pm, about 5 pm to about 80 pm, about 5 pm to about 70 pm, about 5 pm to about 60 pm, about 5 pm to about 50 pm, about 5 pm to about 45 pm, about 5 pm to about 40 pm, about 5 pm to about 35 pm, about 5 pm to about 30 pm, about 5 pm to about 25 pm, or about 5 pm to about 20 pm. In some embodiments, the semi-permeable membrane may be thinner than the mesh. The thickness of the semi-permeable membrane varies depending on several variables such as amount of a sample, a chamber volume, and a chamber size.

[0087] In some embodiments, the semi-permeable membrane extends to the side walls. In certain embodiments, horizontal cross-section of the semi-permeable membrane is in the shape of cross section of the chamber, such as, circle, rectangle, square, oval, rhombus, pentagon, hexagon, nonagon, octagon, or polygon.

[0088] In some embodiments, if horizontal cross-section of the semi-permeable membrane is in shape of circle, the diameter of the semi-permeable membrane is, in some instances, about 5 cm or more, 10 cm or more, 15 cm or more, 20 cm or more, 30 cm or more, 40 cm or more, 40 cm or more, 50 cm or more, 60 cm or more, 70 cm or more, 80 cm or more, 90 cm or more, or 100 cm or more. For example, the diameter of the semi-permeable membrane is, for example, in the range of 5 cm to 500 cm, 5 cm to 400 cm, 5 cm to 300 cm, 5 cm to 200 cm, 5 cm to 150 cm, 5 cm to 100 cm, 5 cm to 90 cm, 5 cm to 80 cm, 5 cm to 70 cm, 5 cm to 60 cm, 5 cm to 50 cm, 5 cm to 40 cm, 5 cm to 30 cm, 5 cm to 25 cm, 5 cm to 20 cm, 7 cm to 20 cm, 8 cm to 20 cm, 10 cm to 20 cm, or 10 cm to 15 cm. The diameter of the semi-permeable membrane varies depending on several variables such as amount of a sample, a chamber diameter, and the like.

[0089] In other embodiments, if horizontal cross-section of the semi-permeable membrane is in shape of non-circle, the width and / or length of the semi-permeable membrane is, in some instances, about 5 cm or more, 10 cm or more, 15 cm or more, 20 cm or more, 30 cm or more, 40 cm or more, 40 cm or more, 50 cm or more, 60 cm or more, 70 cm or more, 80 cm or more, 90 cm or more, or 100 cm or more. The width / length of the semi-permeable membrane varies depending on several variables such as amount of a sample, a chamber width / length, a chamber volume and the like.

[0090] In other embodiments, the semi-permeable membrane is flexible, elastic, or viscoelastic. In some embodiments, the semi-permeable membrane is a cellulose membrane, a polytetrafluoroethylene (PTFE) membrane, or a dialysis membrane.

[0091] In some embodiments, devices of interest may comprise a membrane holder that is configured to hold the semi-permeable membrane in place and provide mechanical support. In certain cases, the membrane holder includes a web, lattice, or cage that includes a relatively inflexible material, such as polystyrene, stainless steel, polyethylene terephthalate, nylon, ABS, polypropylene, polyurethane, polyetherimide, polyphenol sulfide, or another material. The membrane holder may enable the membrane to maintain substantially avoid bulging or buckling during use.

[0092] Hypertonic medium

[0093] Devices of interest include a hypertonic medium. In certain embodiments, the hypertonic medium may be disposed over the semi-permeable membrane such that it is in contact with the semi-permeable membrane. In some embodiments, the hypertonic medium is a sorbent polymer. In certain cases, a sorbent backing is further disposed over the hypertonic medium to provide mechanical support.

[0094] In some embodiments, the hypertonic medium comprises a hypertonic liquid or a hypertonic gel. In certain embodiments, hypertonic medium comprises absorbent polymer, absorbent hydrogel, or absorbent cellulose. In certain embodiments, hypertonic medium is a liquid or gel comprising a solute at a concentration higher than the concentration of the solute in the sample to create a net flow of aqueous phase from the sample to the hypertonic medium. For example, the concentration of the solute in the hypertonic medium may be at least 10X, at least 5X, or at least 2X of the solute in the sample. For example, the concentration of the solute in the hypertonic medium may be greater than or equal to 0.2 grams per milliliter (g / mL). In some embodiments, the concentration of the solute in the solution is less than or equal to 3 g / mL. In some embodiments, the molecular weight of the solute is in a range of 1 ,500 to 1 ,000,000 Daltons (Da). In some embodiments, the solute comprises at least one of a polymer, a detergent, a surfactant, or a micelle. In some embodiments, solute comprises at least one of polyethylene glycol (PEG), polystyrene sulfonate (PSS), polyacrylic acid (PAA), polyethyleneimine (PEI), pectin, or sodium dodecyl sulfate (SDS). In some embodiments, the solute is polyethylene glycol (PEG), e.g., PEG 1500. The PEG), e.g., PEG 1500 may be at a concentration of 2.0 grams per milliliter (g / mL).

[0095] In some embodiments, the hypertonic medium has a thickness in the range of about 1 mm to about 30 cm, such as about 1 mm to about 25 cm, about 1 mm to about 20 cm, about 5 mm to about 15 cm, about 1 cm to about 15 cm, about 1 cm to about 15 cm, about 1 cm to about 10 cm, about 1 cm to about 5 cm, about 1 cm to about 3 cm. In some embodiments, the hypertonic medium is substantially thicker than the mesh or semi-permeable membrane. The thickness of the hypertonic medium varies depending on several variables such as amount of a sample, a chamber volume, and a chamber size. In some embodiments, the hypertonic medium has a volume of at least 1 ml, e.g., 1 ml-30 ml, 1 ml to 20 mL, 5 ml to 50 mL, 5 ml to 25 mL, etc.

[0096] In some embodiments, horizontal cross-section of the hypertonic medium is in the shape of cross section of the chamber, such as, circle, rectangle, square, oval, rhombus, pentagon, hexagon, nonagon, octagon, or polygon.

[0097] In some embodiments, if horizontal cross-section of the hypertonic medium is in shape of circle, the diameter of the hypertonic medium is, in some instances, about 5 cm or more, 10 cm or more, 15 cm or more, 20 cm or more, 30 cm or more, 40 cm or more, 40 cm or more, 50 cm or more, 60 cm or more, 70 cm or more, 80 cm or more, 90 cm or more, or 100 cm or more. For example, the diameter of the hypertonic medium is, for example, in the range of 5 cm to 500 cm, 5 cm to 400 cm, 5 cm to 300 cm, 5 cm to 200 cm, 5 cm to 150 cm, 5 cm to 100 cm, 5 cm to 90 cm, 5 cm to 80 cm, 5 cm to 70 cm, 5 cm to 60 cm, 5 cm to 50 cm, 5 cm to 40 cm, 5 cm to 30 cm, 5 cm to 25 cm, 5 cm to 20 cm, 7 cm to 20 cm, 8 cm to 20 cm, 10 cm to 20 cm, or 10 cm to 15 cm. The diameter of the hypertonic medium varies depending on several variables such as amount of a sample, a chamber diameter, and the like.

[0098] In other embodiments, if horizontal cross-section of the hypertonic medium is in shape of non-circle, the width and / or length of the hypertonic medium is, in some instances, about 5 cm or more, 10 cm or more, 15 cm or more, 20 cm or more, 30 cm or more, 40 cm or more, 40 cm or more, 50 cm or more, 60 cm or more, 70 cm or more, 80 cm or more, 90 cm or more, or 100 cm or more. The width / length of the hypertonic medium varies depending on several variables such as amount of a sample, a chamber width / length, a chamber volume and the like.

[0099] In some embodiments, devices of interest further include a sorbent retainer comprising the hypertonic medium. In other embodiments, the sorbent retainer is filled with the hypertonic medium, such as sorbent polymer. The term “a sorbent retainer” or “a hypertonic medium holder” are used interchangeably throughout the present disclosure. In some embodiments, the sorbent retainer is configured to hold the hypertonic medium in a predetermined shape. In certain cases, the sorbent retainer includes a web, lattice, or cage that includes a relatively inflexible material, such as polystyrene, stainless steel, polyethylene terephthalate, nylon, ABS, polypropylene, polyurethane, polyetherimide, polyphenol sulfide, or another material. The sorbent retainer may cause the hypertonic medium to maintain the predetermined shape. In some embodiments, the sorbent retainer is pre-coated.

[0100] In some embodiments, devices of interest further include a lid in a fitting configuration with respect to the opening of the chamber. In certain cases, the lid comprises a hypertonic medium, a semi-permeable membrane, and a mesh.

[0101] In some embodiments, devices of interest further include a sorbent backing disposed over the sorbent retainer and under the lid.

[0102] In some embodiments, devices of interest may include gaskets and clip retainers.

[0103] In some embodiments, devices of interest further comprise an interfacing unit to receive a sample analysis cartridge. In some embodiments, the device comprises one or more interfacing units to receive one or more sample analysis cartridges. The sample analysis cartridge varies depending on the type of assays and is not limited to the examples described in the present disclosure.

[0104] In some embodiments, devices of interest may combine with a sample analysis cartridge. In some cases, the sample analysis cartridge may be integrated into the device of the present disclosure. In other cases, one or more sample analysis cartridges may be integrated into the device of the present disclosure. Therefore, the concentrate can be automatically transferred to the sample analysis cartridge by pressing the concentrate transfer button on the device.

[0105] FIG. 1 A shows concentration of a target molecule under standard forward osmotic conditions at the membrane surface. As such, the urine sample 101 including target molecule 102 is concentrated by forward osmosis driven by osmotic pressure. As used herein, the term “Forward osmosis (FO)” is a water separation process which uses a semi-permeable membrane and the natural energy of osmotic pressure to separate water from dissolved solutes. The osmotic pressure is used to transport water through the membrane while retaining all the dissolved solutes on the other side. While the conventional forward osmosis uses only passive flow 104 with semi-permeable membrane 106 and osmotic drive absorbent polymer 103, FIG. 1B depicts an operation of the present disclosure that combines osmotic flow-based concentration 104 with a localization process that allows for rapid congregation of the target molecules 102 in the specified area of interest. Capillary forces 105 which is driven by a thin mesh (not shown) are used to drive the molecules 102 towards a target area of interest. This method, hereby referred to as, “mesh-induced-continuous-capillary-flow” leverages the osmotic drive for localization and target concentration.

[0106] FIG. 2A illustrates an exemplary osmotic urine concentration device 200 showing a top lid 203 with an entrance 204 for the urine sample and a base module 201 comprising an outlet (not shown) and a sample extraction region 202. FIG. 2B shows a cross section of the device 200 showing the core osmotic engine that drives target concentration. FIG. 2B further illustrates one possible configuration of the osmotic engine. A larger volume of urine sample in a sample feeder pool 205 feeds into a narrow mesh 209 which shown here as a PP 125pm mesh under a significantly larger osmotic membrane 208 which shown as regenerated cellulose membrane 25 pm thick. 206 indicates sorbent module top. The osmotic drive from the absorbent polymer 207 allows for target concentration in the small volume trapped in the mesh 209 between the membrane 208 and the base module 210, while also driving localization of the target 211 (tuberculosis antigen lipoarabinomannan (LAM) molecules in red). This the osmotic flux drives the localization of target 211 by mesh-lnduced continuous capillary flow between a larger sample reservoir and a thickness-controlled sample layer. Localization of the target molecule 211 is highly sensitive to controlling this flow (among other parameters) without which the target is “lost in the matrix”. Once the concentration reaches a required target, the concentrate can be withdrawn from below via an extraction channel 213. Once the concentration is complete, the concentrate can be removed using different methods to be applied to a diagnostics assay (e.g.: Lateral flow test).

[0107] FIG. 3A shows an exemplary osmotic engine device 300 of the present disclosure according to one embodiment. FIG. 3B shows a cross section of the exemplary device 300 assembled with elements. FIG. 3C shows that the exemplary device 300 comprises (i) a chamber or a testing tray 301 , (ii) a mesh that is seated on top of the chamber (not shown) (iii) clip retainer 302, (iv) module base 303, (v) gasket #2 304, (vi) membrane 305, (vii) sorbent retainer 306 which is pre-coated / filled with sorbent polymer, (viii) sorbent backing 307, (ix) gasket #1 308, (x) module lid 309.

[0108] Samples

[0109] Devices of interest include a sample. As used herein, "sample", "test sample", or "biological sample" refers to a sample containing or suspected of containing an analyte. As used herein, "analyte", "target analyte", and "target molecule" are used interchangeably and refer to any substance being concentrated in the devices disclosed herein. In some embodiments, a sample of the present disclosure is derived from any suitable source. In other embodiments, the sample comprise a liquid, fluent particulate solid, or fluid suspension of solid particles. In certain embodiments, the sample may be a liquid sample or a liquid extract of a solid sample. In some cases, the fluid sample may be concentrated prior to use in an assay. A fluid sample may be concentrated about 1- fold, about 2-fold, about 3-fold, about 4-fold, about 5-fold, about 6-fold, about 10-fold, about 100-fold, or greater, prior to use.

[0110] In some embodiments, the source of the analyte molecule may be synthetic (e.g., produced in a laboratory), the environment (e.g., air, soil, etc.), fluid samples, e.g., water supplies, etc.), an animal, e.g., a mammal, a plant, or any combination thereof.

[0111] In some embodiments, a sample as the source of an analyte is a human bodily substance. The human bodily substance may be a liquid sample or a liquid extract of a solid sample. Non-limiting embodiments of the human bodily substance is bodily fluid, blood, serum, plasma, urine, saliva, sweat, sputum, semen, mucus, lacrimal fluid, tears, dermal fluid, lymph fluid, amniotic fluid, interstitial fluid, intestinal fluid, gastrointestinal fluid, lung lavage, spinal fluid, cerebrospinal fluid, feces, nasal mucus, virginal discharge, tissue, organ, or like.

[0112] In certain embodiments, a sample of the present disclosure is urine. As used herein, the term “urine” refers to a watery, typically yellowish fluid stored in the bladder and discharged through the urethra. Urine is one of the body's chief means of eliminating excess water and salt, and also contains nitrogen compounds such as urea and other waste substances removed from the blood by the kidneys. Collecting a urine sample is well known in the art. In exemplary embodiments, either a "first-catch" or a "mid-stream" sample of urine is collected in a completely sterile container. The first-catch urine sample is the first part of the urine that comes out. The mid-stream urine is for reducing the risk of the sample being contaminated with bacteria from hands, or the skin around the urethra or the tube that carries urine out of the body. In some embodiments, the collected urine sample may be stored in a fridge at 4 °C less than 24 hours in a sealed plastic bag. In certain embodiments, the urine sample is used for infections such as urinary tract infection (UTI), some sexually transmitted infections (STIs) such as chlamydia in men, or kidney damage, such as ACR test.

[0113] In certain embodiments, a sample of the present disclosure is whole blood. Samples for hematology are typically whole blood. The whole blood sample consists of red blood cells, white blood cells, and platelets suspended in a protective yellow liquid known as plasma. In some embodiments, samples for immunoassays and clinical chemistry assays are typically serum or plasma. In some embodiments, the whole blood sample is obtained from a subject. In some embodiments, the subject is a living subject, including an animal and a human. After a sample of whole blood is aspirated from a sample tube, a portion of the sample of whole blood must be removed from the sample of whole blood so that either serum or plasma can be separated from the portion for subsequent use in immunoassay testing or clinical chemistry testing.

[0114] In certain embodiments, a sample of the present disclosure is plasma. As used herein, the term “plasma” refers to the colorless fluid part of blood, lymph, or milk, in which corpuscles or fat globules are suspended. As such, plasma is the blood's liquid component and is made up of water, proteins, waste products, minerals, clotting factors, immunoglobulins, carbon dioxide and hormones. The method for separating plasma from blood is well known in the art. In exemplary embodiments, plasma is produced when whole blood is collected in tubes that are treated with an anticoagulant. The blood does not clot in the plasma tube, thereby the cells are removed by centrifugation. The supernatant, designated plasma is carefully removed from the cell pellet using a Pasteur pipette.

[0115] In certain embodiments, a sample of the present disclosure is serum. As used herein, the term “serum” refers to the watery, clear portion of an animal fluid or plant sap. As used herein, the term “blood serum” refers to an amber-colored, protein-rich liquid that separates out when blood coagulates. In certain embodiments, serum includes, but not limited to, blood serum, serous (or serosal) fluid secreted by the serous glands, and plant sap. The method for separating serum from blood is well known in the art. In exemplary embodiments, the blood serum is collected after whole blood is allowed to clot. The clot is removed by centrifugation, and the resulting supernatant, designated serum, is carefully removed using a Pasteur pipette.

[0116] In certain embodiments, a sample of the present disclosure is a cerebrospinal fluid. The term “cerebrospinal fluid (CSF)” refers to a clear fluid that surrounds and protects the brain and spinal cord. The analysis for cerebrospinal fluid may look for proteins, sugar (glucose), and other substances. The method for collecting cerebrospinal fluid is well known in the art. In exemplary embodiments, cerebrospinal fluid is usually obtained through a lumbar puncture (spinal tap). During the procedure, a needle is inserted usually between the 3rd and 4th lumbar vertebrae and the CSF fluid is collected for testing.

[0117] In certain embodiments, a sample of the present disclosure is saliva. As used herein, the term “saliva” refers to watery liquid secreted into the mouth by glands, providing lubrication for chewing and swallowing, and aiding digestion. Saliva consists of 99% water and 1 % protein and salts. The method of collecting saliva is well known in the art. In some embodiments, saliva sample can be refrigerated for up to a week before it needs to be added to the stabilizing fluid in the tube.

[0118] A wide range of volumes of the fluid sample may be analyzed. In exemplary embodiments, the sample volume is in the range of from 0.1 pL to 1000 mL, such as from 0.1 pL to 100 mL, from 0.1 pL to 50 mL, from 0.1 pL to 30 mL, from 0.1 pL to 10 mL, from 0.1 pL to 5 mL, from 0.1 pL to 3 mL, from 0.1 pL to 1 mL, from 0.1 pL to 900 pL, from 0.1 pL to 800 pL, from 0.1 pL to 700 pL, from 0.1 pL to 600 pL, from 0.1 pL to 500 pL, from 0.1 pL to 400 pL, from 0.1 pL to 300 pL, from 0.1 pL to 200 pL, from 0.1 pL to 100 pL, from 0.1 pL to 90 pL, from 0.1 pL to 80 pL, from 0.1 pL to 70 pL, from 0.1 pL to 60 pL, from 0.1 pL to 50 pL, from 0.1 pL to 40 pL, from 0.1 pL to 30 pL, from 0.1 pL to 20 pL, from 0.1 pL to 10 pL, from 0.1 pL to 5 pL, or the like. In further exemplary embodiments, the sample volume is about 500 pL, 800 pL, 1 ml, 5 ml, 10 ml, 50ml, 100 ml, 500ml, or the like.

[0119] In exemplary embodiments, one or more sample collection devices are, not limited to, syringes, sterile containers, standard urine collection vessels, standard vacutainer urine collection vessels, sponge stick samplers, microsampling devices, micro-needles, or other minimally invasive pain-free blood collection devices; blood collection tube(s); lancets; capillary blood collection tubes; other single fingertip-prick blood collection devices, buccal swabs, nasal / throat swabs, 16-gauge or other size needle, or the like.

[0120] SYSTEM FOR REDUCING SAMPLE VOLUME AND CONCENTRATING AN ANALYTE

[0121] Aspects of the present disclosure include a system for reducing volume of a sample while retaining an analyte of interest in the sample. More specifically, systems of interest comprise the device of the present disclosure and one or more of a sample analysis cartridge, a pipette, and an instruction for reducing volume of a sample and analysis of the sample.

[0122] Systems of interest comprise devices of the present disclosure which are described in the above section.

[0123] Sample analysis cartridge

[0124] Systems of interest further include one or more of a sample analysis cartridge. In certain embodiments, devices of interest comprise an interfacing unit to receive a sample analysis cartridge. In some embodiments, the device comprises one or more interfacing units to receive one or more sample analysis cartridges. The sample analysis cartridge varies depending on the type of assays and is not limited to the examples described in the present disclosure.

[0125] (1) Nucleic Acid analysis cartridge

[0126] Various amplification methods and components will be known to one of ordinary skill in the art and any convenient method can be used in the systems and methods disclosed herein (see, e.g., Zanoli and Spoto, Biosensors (Basel). 2013 Mar; 3(1): 18- 43; Gill and Ghaemi, Nucleosides, Nucleotides, and Nucleic Acids, 2008, 27: 224-243; Craw and Balachandrana, Lab Chip, 2012, 12, 2469-2486; which are herein incorporated by reference in their entirety). Nucleic acid amplification can comprise polymerase chain reaction (PCR), reverse transcription PCR (RT-PCR), quantitative PCR (qPCR), reverse transcription qPCR (RT-qPCR), nested PCR, multiplex PCR, asymmetric PCR, touchdown PCR, random primer PCR, hemi-nested PCR, polymerase cycling assembly (PCA), colony PCR, ligase chain reaction (LCR), digital PCR, methylation specific-PCR (MSP),co-amplification at lower denaturation temperature-PCR (COLD-PCR), allelespecific PCR, intersequence-specific PCR (ISS-PCR), whole genome amplification (WGA), inverse PCR, and thermal asymmetric interlaced PCR (TAIL-PCR). Typically, nucleic acid amplification is employed to increase the number of a target nucleic acid in the sample, e.g., to thereby facilitate detection of the target nucleic acid. As embodied herein, the nucleic acid amplification methods and system components can be configured to amplify a target nucleic acid using any of a variety or combination of suitable amplification techniques.

[0127] (2) Immunoassay cartridge

[0128] In some embodiments, one type of cartridge designed for assaying one or more analytes in a sample is a cartridge for an immunoassay. An immunoassay generally comprises contacting an antigen with an antibody specific for the antigen to form an antibody-antigen complex and detecting the antibody-antigen complex. In some embodiments, the antibody-antigen complex is an antibody-analyte complex. In other embodiments, the analyte is an antigen. In other embodiments, an antigen that may be bound by an antibody includes, but is not limited to, proteins, peptides, polysaccharides, lipids, or nucleic acids.

[0129] Cartridges may be designed to perform various types of immunoassays. In some embodiments, the immunoassay may be a labelled immunoassay. In labelled immunoassays, the antibody-analyte complex may be detected using a detectably labeled antibody. Detectable labels may be selected from a variety of such labels known in the art, but normally are radioisotopes, fluorophores, enzymes (e.g., horseradish peroxidase), or other moieties or compounds which either emit a detectable signal (e.g., radioactivity, fluorescence, color) or emit a detectable signal after exposure of the label to its substrate. Additional labels can include, but are not limited to, DNA probes and reporters, electrochemiluminescent tags, and magnetic particles. Various detectable label / substrate pairs (e.g., horseradish peroxidase / diaminobenzidine, avidin / streptavidin, luciferase / luciferin), methods for labelling antibodies, and methods for using labeled antibodies to detect an antigen are well known in the art. In other embodiments, the immunoassay may be an unlabeled immunoassay. Unlabeled immunoassays are performed without labels and include, but are not limited to, techniques such as immunodiffusion and nephelometry.

[0130] In some embodiments, the immunoassay may be a heterogeneous immunoassay. Heterogeneous immunoassays require separation of the antibody-analyte complex from the other components of the immunoassay prior to analysis. In other embodiments, the immunoassay may be a homogeneous immunoassay. Homogeneous immunoassays do not require separation of the antibody-analyte complex from the other components of the immunoassay prior to analysis.

[0131] In some embodiments, the immunoassay may be a competitive immunoassay. In competitive immunoassays, the analyte competes with a specific quantity of labeled antigen for the antibody. In other embodiments, the immunoassay may be a noncompetitive immunoassay. In noncompetitive immunoassays, excess labeled antibody is used to bind with the analyte.

[0132] Examples of well-known immunoassay variations include, but are not limited to, immunoassay, such as sandwich immunoassay (e.g., monoclonal-polyclonal sandwich immunoassays), enzyme detection, such as enzyme immunoassay (EIA) or enzyme- linked immunosorbent assay (ELISA) (e.g., direct, indirect, competitive, and sandwich ELISA), blotting techniques, lateral flow immunoassay, latex particle agglutination, IgM antibody assay competitive inhibition immunoassay (e.g., forward and reverse), enzyme multiplied immunoassay technique (EMIT), a competitive binding assay, bioluminescence resonance energy transfer (BRET), one-step antibody detection assay, homogeneous assay, heterogeneous assay, capture on the fly assay, Dot blot, Western blot, immunochromatography, immunodiffusion, complement fixation, nucleic acid lateral flow immunoassay (NAFLIA), Fluorescence polarization immunoassay (FPIA), indirect hemagglutination (I HA), indirect immunofluorescence (IIF), direct fluorescent antibody (DFA), rapid enzyme immune assay, immunohistochemistry, immunoblotting, Electrochemiluminsecent immunoassay (ECLIA), Chemiluminescence immunoassay (CLIA), fluorometric assays, immunophenotying, immunomagnetic assays, epithelial immunospot (ELISPOT) assays, immunonephelometric assays, and immunoturbidimetric assays.

[0133] (3) LFAs cartridge

[0134] In certain aspects, the cartridge may be configured for lateral flow assay (LFA). Lateral flow chromatographic immunoassay methods and cartridges have been described extensively. See, e.g., Gordon and Pugh, U.S. Pat. No. 4,956,302; H. Buck, et al., WO 90 / 06511 ; T. Wang, U.S. Pat. No. 6,764,825; W. Brown, et al., U.S. Pat. No. 5,008,080; Kuo and Meritt, U.S. Pat. No. 6,183,972, EP 00987551A3. In one type of LFA cartridge, the test strip is divided into four regions, which can be made of only one kind of material or several kinds of material (e.g., up to four different kinds of materials). The first region is for sample addition. It functions to remove viscous and particulate materials in the sample and also to condition the sample solution for the reactions in the following regions. The second region is a mobile-phase with a color conjugate. The color conjugate may be made from conjugation between a visible color marker (e.g., colored beads, colloidal gold, fluorescent dyes, etc.) and a detection antibody. The detection antibody can bind a specific antigen in the sample (e.g., an analyte or a positive control substance) and forms an antigen-color conjugate complex. The third region of the LFA cartridge is a solid-phase with immobilized capture antibody. The capture antibody can bind the antigen of the antigen-color conjugate complex and forms capture antibody-antigen-color conjugate complex sandwich. The fourth region is for solution absorption. It draws sample solution towards it continuously. In some cases, the analyte(s) and a positive control can be detected on various target lines, respectively, with various reporters. The reporters for each of the various target lines may be the same or different. Examples of suitable reporters include, but are not limited to, visible and fluorescent dyes, latex beads, enzymes, gold nanoparticles, silver nanoparticles, quantum dots, and the like.

[0135] In some cases, when a sample is applied to the diffusion strip of the lateral-flow chromatographic assay cartridge, the liquid in the sample carries the analyte through the diffusion strip in flow direction into an analysis zone where it can be captured by a capture ligand line. In some cases, the LFA cartridge includes a first capture ligand line for capturing a first analyte. In some cases, the LFA cartridge includes a second capture ligand line for capturing a second analyte. In some cases, the LFA cartridge includes first and second capture ligand lines having an amount of the analyte or another material prebound to the diffusion strip of the lateral-flow chromatographic assay cartridge. The reporter may be a diffusible material that can bind to the capture ligand lines in an amount proportional to the amount of bound ligand is present in each line. In response to illumination by the light source, a reporter bound to each of lines provides a signal that can be used to calculate a calibration curves and, in turn, determine the concentration of the analyte in the sample.

[0136] In some cases, the one or more LFA cartridges of the system include an assay region for producing an optically detectable assay result. In some cases, the one or more LFA cartridges of the system include a positive control region in addition to an assay region for producing an optically detectable marking. In some cases, the assay region includes a positive control line for determining that the assay was performed correctly.

[0137] (4) CBC analysis cartridge

[0138] The systems and methods disclosed herein are capable of detecting blood cells or blood cell types. Blood cells and blood cell types that may be detecting by the systems, devices and methods disclosed herein include, without limitation, red blood cells, hemoglobin, white blood cells (including neutrophils, lymphocytes, monocytes, eosinophils, and basophils), platelets, reticulocytes, and nucleated red blood cells. Various measurements of different blood components may be performed, including, but not limited to, cell count, cell size, cell complexity, granularity, hematocrit, mean corpuscular volume, mean corpuscular hemoglobin, and mean corpuscular hemoglobin concentration. In some embodiments, the above disclosed measurements may be performed using stain independent methods in the absence of histological staining.

[0139] (5) Clinical chemistry analysis cartridge

[0140] In certain cases, clinical chemistry may involve detection of electrochemical species or chromogenic reaction product generated by action of an enzyme on a substrate. For example, the substrate may be an analyte present in a sample and the enzyme may be specific for the analyte and may catalytically react with the analyte to generate an electrochemical species ora colored reaction product. In other cases, clinical chemistry may involve capturing the analyte using a first binding member to generate a first complex comprising the analyte and the first binding member; contacting the complex with a second binding member, that binds to the analyte, to generate a second complex comprising the analyte, the first binding member, and the second binding member. The second binding member is conjugated to an enzyme that generates an electrochemical species or chromogenic reaction product upon exposure to a suitable substrate.

[0141] Systems of interest include sample collection devices. The sample collection device is for transporting a concentrated sample from the device to a sample analysis cartridge. In exemplary embodiments, one or more sample collection devices are, not limited to, syringes, sterile containers, standard urine collection vessels, standard vacutainer urine collection vessels, sponge stick samplers, microsampling devices, micro-needles, or other minimally invasive pain-free blood collection devices; blood collection tube(s); lancets; capillary blood collection tubes; other single fingertip-prick blood collection devices, buccal swabs, nasal / throat swabs, 16-gauge or other size needles, or the like. In certain cases, the sample transfer device is a pipette.

[0142] Systems of interest include an instruction for reducing volume of a sample and analysis of the sample. Embodiments shown in FIGs. 4, 6, 8A-8B, 10A-10C, 12A-12B, and 14A-14B encompass exemplary devices and systems of the present disclosure. It is to be understood that this invention is not limited to particular embodiments described herein.

[0143] FIG. 4 illustrates an exemplary system 400 for reducing volume of a sample while retaining an analyte of interest in the sample. The system depicted in FIG. 4 comprises exemplary device 401 of the present disclosure, a standard urine collection vessel 402, a sample analysis cartridge such as LFT 403, and a pipette 404. Device 401 comprises a lid in a fitting configuration with respect to the opening of the chamber.

[0144] FIG. 6 illustrates an exemplary system / device 600 of the present disclosure. In FIG. 6, a sample analysis cartridge such as LFT 602 is integrated into device 600. The urine collection vessel 601 comprises an interfacing unit on the bottom to provide a sample to an inlet of the device 600.

[0145] FIG. 8A illustrates an exemplary standard vacutainer urine collection vessel. FIG. 8B shows an exemplary device / system of the present disclosure which comprises integrated lateral flow test (LFT) cartridge.

[0146] FIG. 10A illustrates an exemplary device / system which comprises integrated a sample analysis cartridge such as LFT and a sponge stick sampler which is configured to insert into the opening of the chamber of the device. FIG. 10B illustrates another exemplary device / system in a horizontal compression shape and a sponge stick sampler. The device is configured to receive a sponge stick sampler on the opening. The sponge stick sampler is inserted horizontally in the device and compressive pressure releases urine. The device depicted in FIG. 10B and FIG. 10C comprises a lid.

[0147] FIG. 12A illustrates an exemplary standard urine collection vessel with an interfacing unit (not shown) on the bottom side. The interfacing unit, as embodied herein, is a break seal. The standard urine collection vessel is placed into the device to break seal and then the sample flows in the device. FIG. 12B illustrates an exemplary device / system of the present disclosure which comprises integrated a sample analysis cartridge such as LFT. The device depicted in FIG. 12B is configured to receive a standard urine collection vessel directly into the opening of the device.

[0148] FIG. 14A illustrates an exemplary standard urine sample collection pot but with septum seal lid and funnel. FIG. 14B illustrates an exemplary device / system of the present disclosure which is in hemisphere shape. The device depicted in FIG. 14B comprises an interface unit to receive a sample analysis cartridge such as LFT. The device depicted in FIG. 14B is configured to receive the standard urine collection pot depicted in FIG. 14A directly into the opening of the device.

[0149] METHOD FOR REDUCING SAMPLE VOLUME AND CONCENTRATING AN ANALYTE

[0150] Aspects of the present disclosure include a method of reducing volume of a sample while retaining an analyte of interest. Methods of interest comprise processing the sample in the devices of the present disclosure or the systems of the present disclosure.

[0151] Methods of interest comprise further steps of providing a sample into the chamber of the device of the present disclosure; processing the sample in the device of the present disclosure; extracting concentrate from a sample extraction region of the device; dispensing the concentrate to a sample analysis cartridge; viewing a result on the cartridge.

[0152] In some embodiments, the providing a sample into the chamber of the device comprises uncovering the opening of the device and pouring the sample into the chamber. In some embodiments, the device of the present disclosure may comprise a lid on top of the chamber such that removing the lid is needed to process the sample. In certain embodiments, the uncovering the opening comprises removing or opening a lid comprising the hypertonic medium, semi-permeable membrane, and mesh.

[0153] In certain embodiments, the device of the present disclosure may not comprise a lid on top of the chamber. In that case, the device may be configured to have a region on top of the device to receive a sample collection vessel. In some embodiments, the region comprises an inlet for entrance of sample into the device. In certain embodiments, the inlet is configured to connect with an interfacing unit of a sample collection vessel to receive a sample without exposure to air. In that case, the sample collection vessel comprises an interfacing unit on the bottom side and the interfacing unit may be a break seal or an air pierceable region. In some embodiments, the inlet comprises a piercing member extending from the inlet and configured for piercing a sample collection vessel pressed against the piercing member. In certain cases, the providing a sample comprises pressing a bottom end or a top end of the sample collection vessel against the piercing member. As such, the inlet of the device, as one embodiment, is configured to puncture the break seal of the sample collection vessel such that the sample can flow into the device. In some embodiments, the providing a sample comprises introducing the sample into an inlet positioned adjacent the opening of the device. In certain cases, the inlet is positioned at a peripheral region of the lid and is fluidically connected to the chamber. In some embodiments, the device comprises a plurality of inlets positioned at peripheral regions of the lid and fluidically connected to the chamber. In certain cases, the inlet is positioned at the center of the lid. In some embodiments, the device comprises a plurality of inlets positioned at the center of the lid and fluidically connected to the chamber.

[0154] In certain embodiments, the providing a sample comprises transferring the sample from a sample collection vessel to the inlet(s) via a sponge stick sampler. In certain embodiments, the providing a sample comprises transferring the sample from a sample collection vessel to the inlet(s) via a Vacutainer®.

[0155] In some embodiments, a sample collection device is used to collect a sample from a subject and provide the sample to the device of the present disclosure. In certain cases, the sample collection device is inserted directly into the device to provide a sample. In certain cases, a sample in a sample collection vessel is poured into the inlet of the device. In exemplary embodiments, one or more sample collection devices are, not limited to, syringes, sterile containers, standard urine collection vessels, standard vacutainer urine collection vessels, sponge stick samplers, microsampling devices, micro-needles, or other minimally invasive pain-free blood collection devices; blood collection tube(s); lancets; capillary blood collection tubes; other single fingertip-prick blood collection devices, buccal swabs, nasal / throat swabs, 16-gauge or other size needles, or the like.

[0156] In some embodiments, methods of interest comprise holding the sample in the device for a period of 5 minutes to 45 minutes to allow for reduction in the volume of the sample. In certain cases, the period for holding the sample in the device is about 5 minutes, about 10 minutes, about 15 minutes, about 20 minutes, about 25 minutes, about 30 minutes, about 35 minutes, about 40 minutes, about 45 minutes, about 50 minutes, about 55 minutes, about 60 minutes or about 150 minutes. The period for holding the sample in the device to allow for reduction in the volume of the sample varies depending on amount of sample volume, test device type, test device size and the like.

[0157] In some embodiments, methods of interest comprise removing or extracting a portion of the reduced volume sample. In certain embodiments, the removing a portion of the reduced volume sample comprises piercing a seal positioned at the sample extraction region. Piercing the seal opens an air seal and allows sample flow from the chamber via the outlet to the extraction channel and removing the reduced volume sample via the pierced seal. In certain embodiments, the removing a portion of the reduced volume sample comprises pipetting the reduced volume sample.

[0158] In some embodiments, methods of interest further comprise operating a sample analysis cartridge. In some cases, the sample analysis cartridge is integrated in the device of the present disclosure. In other cases, the sample analysis cartridge may be inserted into an interfacing unit of the device and operated with the reduced volume sample. The sample analysis cartridge is fluidically connected with the sample extraction channel of the device such that the reduced volume sample can be flown into the sample analysis cartridge.

[0159] In certain cases, the device of the present invention and a sample analysis cartridge are not combined. In that case, the method further comprises removing or extracting the reduced volume sample from the sample extraction region of the device and providing the sample into the sample analysis cartridge by pipetting.

[0160] Embodiments shown in FIGs. 5, 7, 9, 11, 13 and 15 encompass the four core steps needed in the sample (e.g.: urine) processing i.e. , collection, transfer, processing, and dispensing on LFA. It is to be understood that this invention is not limited to particular embodiments described herein.

[0161] FIG. 5 shows an exemplary method of reducing volume of a sample while retaining an analyte of interest. The method comprises (i) sample collection step 501 , (ii) transferring step 502, (iii) processing step 503, (iv) dispensing step 504, and (v) viewing result step 505. More specifically, the method comprises steps of (i) preparing a standard urine collection vessel, collecting urine sample from a subject into the urine collection vessel, and sealing the urine collection vessel (501); (ii) preparing the device of the present disclosure and removing a device lid / cover (502a); removing the urine collection vessel lid (502b); pouring the urine sample into the device (502c); and closing a device lid / cover (502d); (iii) processing (503); (iv) removing or piercing seal tab from a sample extraction region (504a); and extracting concentrate using pipette (504b); (v) dispensing concentrate using pipette to a sample analysis cartridge (LFT) (505a); waiting for 15 minutes; and viewing result on the cartridge (LFT) (505b).

[0162] FIG. 7 shows an exemplary method of reducing volume of a sample while retaining an analyte of interest. The method comprises (i) sample collection step 701 , (ii) transferring step 702, (iii) processing step 703, (iv) dispensing step 704, and (v) viewing result step 705. More specifically, the method comprises steps of (i) preparing a standard urine collection vessel with interfacing unit on the bottom side, collecting urine sample from a subject into the urine collection vessel, and sealing the urine collection vessel (701 ); (ii) placing the urine collection vessel to the device of the present disclosure (702a); and screwing or clicking the urine collection vessel to insert into the device (702b); (iii) processing; waiting about 30 minutes; and observing fluid level and indicator to proceed to next step (703); (iv) pressing concentrate transfer button (704); (v) waiting about 15 minutes; and viewing result on the cartridge (LFT) (705).

[0163] FIG. 9 shows an exemplary method of reducing volume of a sample while retaining an analyte of interest. The method comprises (i) sample collection step 901 , (ii) transferring step 902, (iii) processing step 903, (iv) dispensing step 904, and (v) viewing result step 905. More specifically, the method comprises steps of (i) preparing a standard urine collection vessel; collecting urine sample from a subject into the urine collection vessel (901a); screwing lid on the vessel, wherein the lid is configured to use a vacutainer and sealing the urine collection vessel (901b); and pushing in vacutainer (901c); (ii) removing vacutainer once filled with urine sample (902a); and inserting the vacutainer into device (902b); (iii) processing (903a); waiting about 30 minutes; and observing indicator to proceed to next step (903b); (iv) pressing concentrate transfer button (904); (v) waiting about 15 minutes; and viewing result on the cartridge (LFT) (905).

[0164] FIG. 11 shows an exemplary method of reducing volume of a sample while retaining an analyte of interest. The method comprises (i) sample collection step 1101 , (ii) transferring step 1102, (iii) processing step 1103, (iv) dispensing step 1104, and (v) viewing result step 1105. More specifically, the method comprises steps of (i) preparing a sponge stick sampler; and urinating onto the sponge stick sampler or dipping the sponge stick sampler in urine sample (1101); (ii) placing the sponge stick sampler with sample into device (1102); (iii) processing (1103); and (iv) waiting and observing indicator to proceed to next step (1104); and (v) waiting about 15 minutes; and viewing result on the cartridge (LFT) (1105).

[0165] FIG. 13 shows an exemplary method of reducing volume of a sample while retaining an analyte of interest. The method comprises (i) sample collection step 1301 , (ii) transferring step 1302, (iii) processing step and dispensing step 1303, and (iv) viewing result step 1304. More specifically, the method comprises steps of (i) preparing a standard urine collection vessel with interfacing unit on the bottom side, collecting urine sample from a subject into the urine collection vessel, and sealing the urine collection vessel (1301); (ii) placing or inserting the urine collection vessel into the device to break seal (1302a); and starting the device running (1302b); (iii) processing; waiting about 45 minutes for device to process sample, wherein upper body of the device is turning during the process (1303a, 1303b); results ready when the device has reached end of travel (1303c); and (iv) viewing result on the cartridge (LFT) (1304).

[0166] FIG. 15 shows an exemplary method of reducing volume of a sample while retaining an analyte of interest. The method comprises (i) sample collection step 1501 , (ii) transferring step 1502, (iii) processing step 1503, (iv) dispensing step 1504, and (v) viewing result step 1505. More specifically, the method comprises steps of (i) preparing a standard urine collection pot but with septum seal lid and funnel; screwing the funnel onto a sample tube (1501a); urinating into the funnel (1501 b); unscrewing the sample tube from the funnel (1501 c); and screwing lid onto the sample tube (1501 d); (ii) inserting the sample tube into the device (1502); (iii) processing; waiting about 30 minutes; and observing indicator to proceed to next step (1503); (iv) pressing button on a sample analysis cartridge (LFT) (1504); (v) waiting about 15 minutes; and viewing result on the cartridge (LFT) (1505).

[0167] DIAGNOSTIC ASSAYS

[0168] Once the concentration is complete, the concentrated sample can be removed from the device of the present disclosure using different methods to be applied to a diagnostics assay. Aspects of systems and methods of the present disclosures involve the analysis of analytes present in the concentrated sample. The analysis of analytes may comprise segregating the analyte(s) into one or more sample detection regions in devices and analyzing the analyte(s). Segregation of the sample and / or the analyte(s) contained therein may occur through the movement of fluids containing the sample and / or the analyte(s) contained therein or the movement of beads, nanobeads, or microparticles that bind to the sample or analyte(s) contained therein into sample detection regions. In some embodiments, the analysis comprises qualitative detection. In some embodiments, the analysis comprises quantitative detection. In some embodiments, the analysis comprises multiplexing analysis. In some embodiments, the analysis comprises multi-parallel analysis. In some embodiments, the sample analysis is nucleic acid sequencing. In some embodiments, the nucleic acid sequencing is via nanopore translocation and analysis.

[0169] In some cases, assays of the present disclosure can be used to determine the presence or absence of an analyte in a sample or measure the amount of an analyte in a sample to identify or assess a disease or condition. Measurements of an analyte can be used, for example, but not by way of limitation, determine the likelihood of developing a disease or condition; diagnose, identify, or classify a disease or condition; estimate prognosis; determine the extent of a disease or condition; determine appropriate treatment; predict response of a disease or condition to treatment; monitor response of a disease or condition to treatment; determine treatment efficacy; and identify recurrence of a disease or condition.

[0170] In exemplary embodiments, systems of interest and methods of interest uses various assay cartridges, such as nucleic acid analysis cartridges, immunoassay cartridges, LFA cartridges, CBC analysis cartridge, Clinical chemistry analysis cartridge, and the like, which are described in the present disclosure.

[0171] UTILITY

[0172] The subject devices, systems, and methods described in this disclosure find use in a variety of applications where it is desirable to concentrate or enrich target molecules from an aqueous solution while allowing for specified localization. The present disclosure provides a passive force osmotic-pressure driven system that drives concentration and localization of a target molecule by combining Forward Osmosis with Localization of Target by Mesh-Induced Continuous Capillary Flow. The present disclosure provides the use of a mesh to support semi-permeable membrane and maintain a very narrow sample gap that enables flow-based target localization. The present disclosure provides continuous flow-based concentration.

[0173] In some aspects, devices, systems, and methods of interest use a disc-shape form factor allowing converging flow towards the center leading to an increase in the concentration factor. In other aspects, other form factors can also be leveraged to allow specific localization of a target in a device. In some embodiments, devices, systems, and methods of interest leverages a mesh-based support (i) to prevent formation of uneven wet membrane surface (common to RC membranes) which can lead to formation of local pockets of sample (when not reinforced with the mesh) and (ii) driven the localization of the target by capillary-assisted flow. Localization of the sample in such spaces (especially in very small final concentrate volumes where capillary forces take over) can lead to significant or near complete loss of target.

[0174] In one aspect, devices, systems, and methods of interest can be used for collection and processing of urine or other bio-samples towards a bioassay for lateral flow detection. In another aspect, devices, systems, and methods of interest can be used for collection and processing for concentration of urine samples towards sample preparation of bioassays or other bioanalytical processes. In still another aspect, devices, systems, and methods of interest can be used for collection and processing of other biological liquid samples, such as saliva, sputum, cerebrospinal fluid or blood among others towards a bioassay and / or sample preparation for other bioanalytical processes. In yet another aspect, devices, systems, and methods of interest can be used for collection and concentration of target molecules in a sample that can additionally use magnetic separation (immunoprecipitation among others; i.e. the particles concentrate the target in the already high concentration osmotic concentrate and the bound target can then be removed from the concentrator to be eluted by a solution to resuspend the target molecule for downstream detection). In further aspect, devices, systems, and methods of interest can be used for purification and / or concentration of target molecules from samples solution.

Claims

CLAIMSWE CLAIM:1 . A device for reducing volume of a sample while retaining an analyte of interest in the sample, the device comprising: a chamber comprising side walls defining an opening; a mesh comprising a plurality of capillary channels disposed over the opening; a semi-permeable membrane disposed over the mesh and comprising pores permeable to water and substantially impermeable to the analyte; and a hypertonic medium in contact with the semi-permeable membrane, wherein the device is configured for generating: a first concentration force driven by osmotic flow of water from a sample present in the chamber to the hypertonic medium across the mesh and the semi- permeable membrane, thereby reducing the volume of the sample, and a second concentration force driven by a mesh-induced capillary action, wherein the second concentration force is orthogonal to the first concentration force, thereby moving solutes present in the sample towards a center of the chamber.

2. The device of claim 1 , comprising an outlet positioned at a substantially central region of the device opposite the opening.

3. The device of claim 1 , wherein the outlet comprises a closed configuration and an open configuration.

4. The device of claim 3, wherein the closed configuration comprises an air seal.

5. The device of claim 3 or claim 4, wherein the closed configuration comprises a pierceable seal.

6. The device of claim 3 or claim 4, wherein the closed configuration comprises a diaphragm.

7. The device of any one of claims 3-6, wherein the outlet is flu idically connected to an extraction channel, wherein the extraction channel extends from the outlet to a sample extraction region accessible from the exterior of the device.

8. The device of any one of claims 1 -7, comprising an inlet for entrance of the sample into the chamber, wherein the inlet is positioned adjacent the opening.

9. The device of any one of claims 1-8, wherein the mesh is a polypropylene mesh, a nylon mesh, or a polypropylene / nylon mesh.

10. The device of any one of claims 1-9, wherein the mesh has a thickness in the range of about 50 pm to about 200 pm.

11. The device of any one of claims 1-10, wherein the semi-permeable membrane is cellulose membrane or a polytetrafluoroethylene (PTFE) membrane.

12. The device of any one of claims 1-11 , wherein the hypertonic medium comprises a hypertonic liquid or a hypertonic gel.

13. The device of any one of claims 1-12, wherein the hypertonic medium comprises absorbent polymer, absorbent hydrogel, or absorbent cellulose.

14. The device of any one of claims 1-13, wherein the chamber is in shape of cylinder, conical frustum, cuboid, cube, rectangular prism, triangular prism, pentagonal prism, or truncated pyramid.

15. The device of any one of claims 1-14, further comprising a lid in a fitting configuration with respect to the opening of the chamber.

16. The device of any one of claims 1-15, further comprising a sorbent retainer comprising the hypertonic medium.

17. The device of any one of claims 1 -16, comprising a sorbent backing disposed over the sorbent retainer and under the lid.

18. The device of any one of claims 1-17, further comprising the sample.

19. The device of claim 18, wherein the sample is a biological sample, an environmental sample, or a synthetic sample.

20. The device of claim 19, wherein the sample is a biological sample and comprises urine, blood, serum, plasma, saliva, sweat, sputum, semen, mucus, lacrimal fluid, lymphatic fluid, amniotic fluid, interstitial fluid, gastrointestinal fluid, lung lavage, cerebrospinal fluid, or vaginal discharge.21 . A system for reducing volume of a sample while retaining an analyte of interest in the sample, the system comprising: the device of any one of claims 1-20 and one or more of: a sample analysis cartridge; a pipette; an instruction for reducing volume of a sample and analysis of the sample.

22. A method of reducing volume of a sample while retaining an analyte of interest, the method comprising processing the sample in the device of any one of claims 1-20 or the system of claim 21.

23. The method of claim 22, comprising providing a sample into the chamber of the device.

24. The method of claim 23, wherein the providing a sample comprises uncovering the opening of the device and pouring the sample into the chamber.

25. The method of claim 24, wherein the uncovering the opening comprises removing a lid comprising the hypertonic medium, semi-permeable membrane and mesh.

26. The method of claim 23, wherein the providing the sample comprises introducing the sample into an inlet positioned adjacent the opening of the device.

27. The method of claim 26, wherein the inlet is positioned at a peripheral region of the lid and is fluidically connected to the chamber.

28. The method of claim 26, wherein the device comprises a plurality of inlets positioned at peripheral regions of the lid and fluidically connected to the chamber.

29. The method of any one of claims 23-28, wherein providing the sample comprises transferring the sample from a sample collection vessel to the inlet(s) via a sponge stick sampler.

30. The method of any one of claims 26-29, wherein the inlet comprises a piercing member extending from the inlet and configured for piercing a sample collection vessel pressed against the piercing member and providing the sample comprises pressing a bottom end or a top end of the sample collection vessel against the piercing member.31 . The method of any one of claims 25-27, wherein providing the sample comprises transferring the sample from a sample collection vessel to the inlet(s) via a Vacutainer®.

32. The method of any one of claims 21-30, comprising holding the sample in the device for a period of 5 minutes to 45 minutes to allow for reduction in the volume of the sample.

33. The method of claim 31 , comprising removing a portion of the reduced volume sample.

34. The method of claim 32, wherein removing a portion of the reduced volume sample comprises piercing a seal positioned at the sample extraction region, wherein piercing the seal opens an air seal and allows sample flow from the chamber via the outlet to the extraction channel and removing the reduced volume sample via the pierced seal.

35. The method of claim 32 or 33, wherein removing a portion of the reduced volume sample comprises pipetting the reduced volume sample.

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