Devices and methods for multi-step, integrated sample processing

Abstract

Described herein are devices for processing samples to enrich an analyte. In some cases, the device comprises: a sample chamber having a bottom comprising a pierceable element, the sample chamber fluidically connected through the pierceable element to a reaction chamber, a piercing element that pierces the pierceable element, a column fluidically connected to the reaction chamber, wherein the column comprises a bed that binds constituents of the sample other than the analyte thereby producing an analyte enriched sample, and a collection element that collects the analyte enriched sample. Methods of using the devices disclosed herein to enrich an analyte in a sample are also provided. In certain embodiments, the sample is urine.

Description

DEVICES AND METHODS FOR MULTI-STEP, INTEGRATED SAMPLE PROCESSINGCROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims benefit under 35 U.S.C. § 119(e) of provisional application serial No. 63 / 703,445, filed October 4, 2024, which application is hereby incorporated by reference in its entirety.INTRODUCTION

[0002] Processing biological samples, such as urine, often requires pre-treatment and conditioning of the sample in a single or multi-step processing. Devices that allow such processing of biological samples can facilitate appropriate handling of biological samples.SUMMARY

[0003] Certain aspects of the disclosure provide devices that can facilitate easier and faster processing for detection, isolation, or enrichment of target analytes from biological samples. Improved sample processing can also enhance detection capability and provide for minimal user involvement or interactions. Reduced user interaction with sample processing also mitigates chemical and biohazard risks, such as contamination and allow for traceability of samples. Moreover, sample processing devices can allow automation and better control over timing of different processing steps. In certain aspects, the disclosure provides devices for integrated handling of a multi-step process of biological samples, such as urine. These multi-step process provides for accurate metering, time and user control transfer of reagents, incubation with reagents, purification, and collection of processed samples. Thus, using the devices disclosed herein, a user can carry out sample processing steps with minimal interaction with the devices, biological sample, and / or reagents.

[0004] The devices disclosed herein provide certain advantages. For example, in some cases, the devices do not require a power source or specialized instrumentation and, hence, are suitable for use in low resource situations. Also, several processing steps that usually require multiple containers and user interactions are performed in one device with minimal user interaction. A user typically does not need to monitor timing of several steps. For example, in certain embodiments, a user is given a visual clue about completion of a step and need for further user involvement. Moreover, in some cases, a processed sample is ready to use for analyte detection and a user does not need to measure the final volume, further avoiding errors.The transfer steps can be within the device and, hence, are reproducible and provide higher reliability of sample processing. Furthermore, with minimal user interaction, the devices disclosed herein remove user errors and variability thereby providing accurate and reproducible results.

[0005] The devices disclosed herein allow metering of samples, time controlled transfer of a sample to a first incubation step, transfer of the incubated sample to a purification column, for example, via inversion of the device or a movement of a piston bore, processing of the sample in the purification column, and collection of the processed sample in a collection element. The sample so collected can be used in a detection method, such as in a lateral flow immunoassay.

[0006] Thus, certain aspects of the disclosure provide a sample processing and analyte enriching device. In some cases, the device comprises:

[0007] a sample chamber having a bottom comprising a pierceable element, the sample chamber fluidically connected through the pierceable element to a reaction chamber,

[0008] a piercing element that pierces the pierceable element,

[0009] a column fluidically connected to the reaction chamber, wherein the column comprises a bed that binds constituents of the sample other than the analyte thereby producing an analyte enriched sample, and

[0010] a collection element that collects the analyte enriched sample.

[0011] In certain embodiments, the sample chamber and the column are located on the same side of the reaction chamber. In further embodiments, the column is located in between the reaction chamber and the collection element.

[0012] In some cases, a sample processing and analyte enriching device, when in operation, comprises:

[0013] i) in a sample loading position: a sample chamber comprising a bottom comprising a pierceable element, a piercing element that, when actuated, pierces the pierceable element thereby causing the sample loaded into the sample chamber to flow under gravity into a reaction chamber, and

[0014] ii) in an analyte enrichment position: a column fluidically connected to the reaction chamber such that the sample from the reaction chamber flows through the column under gravity, wherein the column comprises a bed that binds constituents of the sample other than the analyte thereby producing an analyte enriched sample, wherein the analyte enriched sample collects in a collection element,

[0015] wherein the device in the analyte concentration position is in a physically inverted orientation as compared to the device in the sample loading position.

[0016] In further aspects, a sample processing and analyte enriching device comprises:

[0017] a sample chamber fluidically connected through a channel to a reaction chamber, wherein the channel allows a timed transfer of a sample from the sample chamber to the reaction chamber,

[0018] a column fluidically connected to the reaction chamber, wherein the column comprises a bed that binds constituents of the sample other than the analyte thereby producing an analyte enriched sample, and

[0019] a collection element that collects the analyte enriched sample.

[0020] In even further aspects, the disclosure provides a method for processing a sample to enrich an analyte, the method comprising:

[0021] loading a sample in a sample chamber of a device, the sample chamber comprising a bottom comprising a pierceable element,

[0022] actuating a piercing element to pierce the pierceable element thereby causing the sample loaded into the sample chamber to flow under gravity into a reaction chamber,

[0023] incubating the sample in the reaction chamber,

[0024] inverting the device to allow the sample from the reaction chamber to flow through a column that is fluidically connected to the reaction chamber, wherein the column comprises a bed that binds constituents of the sample other than the analyte, and wherein the sample from the reaction chamber flows through the column under gravity to produce an analyte enriched sample.BRIEF DESCRIPTION OF THE FIGURES

[0025] FIGS. 1A-1B describe an exemplary device. FIG. 1A provides a schematic representation of the exemplary device and FIG. IB provides a photograph of the exemplary device depicted in FIG. 1A.

[0026] FIGS. 2A-2D depict operation of the exemplary device of FIGS. 1A-1B.

[0027] FIGS. 3A-3B depict an exemplary device.

[0028] FIGS. 4A-4D depict an exemplary device as well as the operation of the device through various steps.

[0029] FIGS. 5A-5D depict an exemplary device as well as the operation of the device through various steps.

[0030] FIGS. 6A-6E depict an exemplary device as well as the operation of the device through various steps up to the preparation of a sample with enriched target analyte.

[0031] FIGS. 7A-7C depict various exemplary options for sample collection and detection using the exemplary devices of FIGS. 6A-6E.

[0032] FIGS. 8A-8D depict an exemplary device as well as the operation of the device through various steps.DETAILED DESCRIPTION

[0033] Certain aspects of the present disclosure provide devices for multi-step processing of biological samples, such as urine. Certain other aspects of the disclosure provide methods of processing biological samples, such as urine, using the devices disclosed herein. The devices and methods disclosed herein allow faster processing of biological samples with minimal user interaction such that the processing is simplified and the risk is reduced of chemical and biological contamination typically caused by conventional methods.

[0034] Before the present devices 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 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.

[0035] 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 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 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 devices and methods.

[0036] 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.

[0037] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. In case of conflict, the present document, including definitions, will control. Preferred methods and materials are described below, although methods and materials similar or equivalent to those described herein can be used in practice or testing of the present disclosure. All publications, patent applications, patents and other references mentioned herein are incorporated by reference in their entirety. The materials, methods, and examples disclosed herein are illustrative only and not intended to be limiting.

[0038] 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.

[0039] The terms "comprise(s)," "include(s)," "having," "has," "can," "contain(s)," and variants thereof, as used herein, are intended to be open-ended transitional phrases, terms, or words that do not preclude the possibility of additional acts or structures. The singular forms "a," "an" and "the" include plural references unless the context clearly dictates otherwise. The present disclosure also contemplates other embodiments "comprising," "consisting of" and "consisting essentially of," the embodiments or elements presented herein, whether explicitly set forth.

[0040] The term "comprising" is used herein as requiringthe 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 components 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).

[0041] 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.

[0042] Forthe recitation of numeric ranges herein, each intervening numberthere between with the same degree of precision is explicitly contemplated. For example, for the range of 6-9, the numbers 7 and 8 are contemplated in addition to 6 and 9, and for the range 6.0-7.0, the number 6.0, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9, and 7.0 are explicitly contemplated.

[0043] All ranges disclosed herein are inclusive of the recited endpoint and independently combinable (for example, the range of "from 2 to 10" is inclusive of the endpoints, 2 and 10, 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.

[0044] 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.

[0045] It should be noted that many of the terms used herein are relative terms. For example, the terms "upper" and "lower" are relative to each other in location, i.e., an upper component is located at a higher elevation than a lower component in a given orientation, but these terms can change if the component is flipped. 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 through the outlet out of the structure.

[0046] The terms "horizontal" and "vertical" are used to indicate direction relative to an absolute reference, i.e., ground level. However, these terms should not be construed to require structures to be absolutely parallel or absolutely perpendicular to each other. For example, the first vertical structure and the second vertical structure are not necessarily parallel to each other. The terms "top" and "bottom" are used to 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 "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.

[0047] 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.

[0048] 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. Thecitation 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.

[0049] 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.

[0050] 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 devices and methods. Any recited method can be carried out in the order of events recited or in any other order which is logically possible.DEVICES

[0051] As summarized above, certain aspects of the present disclosure provide sample processing and analyte enriching devices.

[0052] In some embodiments, a sample processing and analyte enriching device comprises:

[0053] a sample chamber having a bottom comprising a pierceable element, the sample chamber fluidically connected through the pierceable element to a reaction chamber,

[0054] a piercing element that pierces the pierceable element,

[0055] a column fluidically connected to the reaction chamber, wherein the column comprises a bed that binds constituents of the sample other than the analyte thereby producing an analyte enriched sample, and

[0056] a collection element that collects the analyte enriched sample.

[0057] The terms "fluidic connection," "fluidically connected," and the like as used herein indicate that the two components of a device are directly or indirectly connected so that a fluid present in one component can be transferred to the other component. For example, a sample chamber is fluidically connected to a reaction chamber indicates that the sample chamber can have a direct fluidic connection to the reaction chamber or intervening connections or chambers.

[0058] The term "pierceable element" as used herein refers to a region of a bottom of a sample chamber that can be pierced by a piercing element in the device. The piercing element can be pierced using a piercing element with minimal efforts by a user, for example, a force applied onthe piercing element by a finger push that is similar to the force applied in turning on an electric switch.

[0059] Accordingly, in some cases, the pierceable element can be made from a readily breakable plastic film or an aluminum foil. Any other material suitable for such use can be used in the devices disclosed herein and such embodiments are within the purview of the disclosure.

[0060] In some cases, a sample chamber comprises a bottom that is partially pierceable, i.e., only a portion of the bottom is made from a pierceable material. Alternatively, a sample chamber can comprise a bottom that is entirely pierceable, i.e., the bottom of the sample chamber is entirely made from a pierceable material.

[0061] A piercing element pierces the pierceable element in the floor of the sample chamber. A piercing element can comprise a pointed tip that, when moved with sufficient force, pierces the pierceable element.

[0062] In some cases, the piercing element is movable between a non-piercing position and a piercing position, wherein in the non-piercing position the piercing element does not touch the pierceable element and in the piercing position the piercing element pierces the pierceable element. Alternatively, in some cases, the piercing element is stationary and the sample chamber is movable between a non-piercing position and a piercing position. In the non-piercing position, the piercing element does not touch the pierceable element and in the piercing position the piercing element pierces the pierceable element.

[0063] In some cases, a piercing element is operably connected to a piercing actuation element that moves the piercing element from a non-piercing position to a piercing position. In some cases, a piercing actuation element can be moved by a user to move an operably connected piercing element from a non-piercing position to a piercing position. In some cases, a piercing actuation element can be a lid of the sample chamber, which when moved to close the sample chamber also moves the piercing element from the non-piercing position to the piercing position.

[0064] When the pierceable element is pierced, a sample loaded in the sample chamber transfers under the force of gravity into the reaction chamber. In some cases, the fluidic connection between the sample chamber and the reaction chamber is a timer channel.

[0065] A "timer channel" as used herein refers to a channel or fluidic connection that provides a certain fluidic resistance and allows a fluid to move from one end of the channel to the other at a desirable rate. For example, if 1 ml of urine sample is added to the sample chamber and a user desires that the 1 ml sample chamber is added to the reaction chamber in 2 mins, thedimensions and structure of a timer channel is such that the urine from the sample chamber transfers into the reaction chamber at the rate of 0.5 ml / min. As a skilled artisan can envision, the functionality of the timer channel depends on the dimensions of the channel, material used to make the channel, as well as the viscosity of the sample to be analyzed.

[0066] In certain aspects, instead of a sample chamber having a bottom having a pierceable element, the devices disclosed herein comprise a sample chamber having a timer channel that fluidically connects the sample chamber to the reaction chamber. An example of a device comprising such timer channel is provided in FIG. 6A.

[0067] In certain embodiments, in a device disclosed herein, the sample chamber and the column are located on the same side of the reaction chamber. For example, when the device is oriented such that the reaction chamber is towards the bottom of the device, the sample chamber as well as the column are above the reaction chamber. Thus, in this orientation, a sample loaded in a sample chamber can move under the force of gravity into the reaction chamber; however, the sample from the reaction chamber does not transfer through the column. Examples of devices in such orientation are provided in FIGS. 1A-1B, 2A, 3A-3B, 4A, 5A, and 6A.

[0068] The same device when oriented such that the reaction chamber is towards the top of the device, the sample chamber as well as the column are below the reaction chamber. Thus, in this orientation, a sample loaded in a reaction chamber can move under the force of gravity through the column. Examples of devices in such orientation are provided in FIGS. 2D, 4D, 5C-5D, and 6D-6E.

[0069] Accordingly, certain aspects of the disclosure provide a sample processing and analyte enriching device that, when in operation, comprises:

[0070] i) in a sample loading position: a sample chamber comprising a bottom comprising a pierceable element, a piercing element that, when actuated, pierces the pierceable element thereby causing the sample loaded into the sample chamber to flow under gravity into a reaction chamber, and

[0071] ii) in an analyte enrichment position: a column fluidically connected to the reaction chamber such that the sample from the reaction chamber flows through the column under gravity, wherein the column comprises a bed that binds constituents of the sample other than the analyte thereby producing an analyte enriched sample, wherein the analyte enriched sample collects in a collection element,

[0072] wherein the device in the analyte concentration position is in a physically inverted orientation as compared to the device in the sample loading position.

[0073] In some cases, the reaction chamber comprises one or more reagents that process the sample to render it suitable for analyte analysis. In an exemplary embodiment, the one or more reagents can comprise a surfactant that increases the availability of an analyte in the sample. For example, a surfactant can increase the availability of lipoarabinomannan (LAM) in a urine sample so that LAM can be detected in the surfactant treated urine sample.

[0001] The surfactant that can be used in the devices disclosed herein can be any suitable surfactant, including but not limited to, a non-ionic surfactant, an anionic surfactant, a cationic surfactant, an amphoteric surfactant, or a zwitterionic surfactant.

[0002] A non-ionic surfactant has no ionic charge. Certain non-limiting examples of nonionic surfactants include alcohol ethoxylates, nonylphenoxy polyethylenoxy alcohols, ethylene oxide / propylene oxide block copolymers, polyethylene glycol hexadecyl ether, and polysorbate 20.

[0003] An anionic surfactant has negatively charged hydrophilic end of the molecule. The negatively charged part of an anionic molecule can be sulfonates, sulfates, or carboxylates. These are usually neutralized by positively charged metal cations such as sodium or potassium. Non-limiting examples of anionic surfactants include sodium dodecyl sulfate, sodium alkylbenzene sulfonates, sodium stearate, and potassium alcohol sulfates.

[0004] A cationic surfactant has a positively charged hydrophilic end of the molecule. The positively charged end is typically derived from nitrogen compounds. Non-limiting examples of cationic surfactants include Cs to Cio alkyl hydroxyethyl dimethylammonium chloride and Cs to Cio alkylamidodimethyl propylamine.

[0005] An amphoteric surfactant can change its charge depending on the pH of the medium. Therefore, an amphoteric surfactant can be a cationic, anionic, or non-ionic surfactant depending on the pH. Certain non-limiting examples of amphoteric surfactants include alkylamidopropylamine N-oxide (APAO), alkyldimethylamine N-oxide (AO), alkylbetaine (Bt), alkylamidopropylbetaine (APB), cocamidopropyl betaine, cocoamphoacetate, and cocoa mphodiacetate.

[0006] A zwitterionic surfactant Zwitterionic surfactants has two distinct and opposite charges on the molecule at either adjacent or non-adjacent sites. The presence of both positive and negative charges renders the molecule overall neutral at neutral pH. Non-limiting examples of zwitterionic surfactants include betaines, such as laurylamidopropyldimethylbetaine;sulfobetaines, such as lauryl hydroxysultaine and myristyl sulfobetaine; amine oxides, such as lauryl dimethylamine oxide; amido betaines; and imidazoline betaines.

[0007] In certain embodiments, the surfactant used in the methods disclosed herein is: sodium dodecyl sulfate and its variants, polyoxyethylenesorbitan monolaurate and its variants, polyethylene glycol hexadecyl ether, alcohol ethoxylate, secondary alcohol ethoxylate and its variants, myristyl sulfobetaine and its variant and the like.

[0008] In a specific embodiment, the surfactant used in the methods disclosed herein is: myristyl sulfobetaine, polyethylene glycol hexadecyl ether, polysorbate 20, secondary alcohol ethoxylate - 9, secondary alcohol ethoxylate - 7, sodium dodecyl sulfate, alcohol ethoxylate - 9, or a combination thereof.

[0074] After a sample is appropriately treated with one or more reagents in the reaction chamber, a device disclosed herein is inverted by a user. A user can be provided a visual clue or indication that the sample treatment in the reaction chamber is complete.

[0075] Once a device disclosed herein is inverted so that the reaction chamber is above the column, the treated sample flows through the column. The column comprises a bed that binds constituents of the sample other than the analyte thereby producing an analyte enriched sample.

[0076] In certain embodiments, the treated sample rapidly flows through the column. Thus, a sample can flow through the column in less than 60 seconds, particularly, less than 50 seconds, such as less than 40, 30, 20, 10, or 5 seconds.

[0077] Various alternatives of a column bed that binds certain constituents of a sample other than the analyte are well known in the art and such embodiments are within the purview of the disclosure.

[0078] In an exemplary embodiment, the bed comprises beads that bind to constituents other than the analyte. Such beads can comprise an inert substrate coated with binding material that binds to constituents other than the analyte.

[0079] When the sample travels through the column, the column bed binds to constituents other than the analyte and an analyte enriched sample is collected in a collection element.

[0080] In some cases, a device disclosed herein provides a visual clue to a user that an analyte enriched sample is collected in the collection element. This may include a transparent window through which a user can see the collected sample. A visual clue may be a color change in a collection element indicative of a sample collection. Once a user is provided such clue, the usercan proceed with further method steps, such as removing the collection element and analyzing the collected analyte enriched sample.

[0081] In some cases, a collection element is a collection chamber. A collection element can also comprise, with or without a collection chamber, one or more capillary channels that absorb an analyte enriched sample.

[0082] In some cases, the collection element is removable from the rest of the device. The term "removable from the rest of the device" as used herein indicates that the collection element can be separated from the rest of the device without structurally damaging any component of the device. In some cases, the collection element can operably engage with elements or structures on the rest of the device such that the collection element can be removed by disengaging the elements. For example, a collection element can be "clicked out" of an engaging structure on the device so that the collection element can be removed without structurally damaging any component of the device.

[0083] The collection element so separated from the rest of the device can be used to analyze the analyte enriched sample.

[0084] In some cases, an analyte enriched sample can be analyzed in an assay device that receives the analyte enriched sample from a fluidic connection to the collection element of the device. Certain such embodiments are described in FIG. 7B and are discussed in Example 5.

[0085] In some cases, the fluidic connection between the reaction chamber and the column comprises a dissolvable block, such as a dissolvable film, that dissolves at a certain rate thereby opening after a predetermined time a fluidic connection between a reaction chamber and a column. In some cases, a dissolvable block comprises poly vinyl alcohol (PVA) film that is engineered to release a fluid from a reaction chamber into a column in a certain time range. A dissolvable block can be designed to allow movement of a sample from a reaction chamber into a column over 1 second to about 2 mins or more. Such dissolvable block could be used for aqueous solutions as well as alcoholic solutions, such as ethanol based solutions. In further embodiments, the dissolvable block requires application of heat before it dissolves.

[0086] Thus, in certain embodiments where such dissolvable block is used in a reaction chamber, after a sample is loaded into a reaction chamber, the device is inverted. The dissolvable block prevents the movement of sample from the reaction chamber into the column. As the dissolvable block dissolves over a predetermined time, the reagents in the reagent chamber act upon the sample. Around the time the dissolvable block dissolves and creates a fluidic connection between the reaction chamber and the column, the reagents react with the sampleappropriate ly. As the dissolvable block dissolves, the reagent treated sample flows through the column and collects in a collection element.

[0087] In further embodiments, similar dissolvable block can also be present in the fluidic connection between the column and the collection element. Thus, a sample that passes through the column is held within the column before it transfers to the collection element.Methods

[0088] The devices disclosed herein can be used for processing a biological sample, for example, a urine sample for enriching an analyte.

[0089] The term "biological sample" as used herein includes any sample comprising non-cellular constituents. In some embodiments, a biological sample is a biological fluid sample. A suitable biological sample processed in the devices disclosed herein can be blood, urine, saliva, sweat, sputum, semen, mucus, lacrimal fluid, lymph fluid, amniotic fluid, interstitial fluid, lung lavage, cerebrospinal fluid, feces, or the like. A biological sample can also be a slurry of an otherwise solid tissue, such as a liver or a kidney, homogenized in a buffer to prepare a slurry comprising cells and non-cellular constituents. A biological sample can also comprise cultured cells in a suitable buffer. Additional examples of biological samples that could be processed in the devices disclosed herein can be readily identified by a person of ordinary skill in the art and such embodiments are within the purview of the disclosure.

[0090] A biological sample can be obtained from any suitable subject. "Subject" as used herein refers to any animal, including, but not limited to, a mammal (e.g., cow, pig, camel, llama, horse, goat, rabbit, sheep, hamsters, guinea pig, cat, dog, rat, and mouse, a non-human primate (for example, a monkey, such as a cynomolgus or rhesus monkey, chimpanzee, etc.) and a human). In some embodiments, the subject may be a human or a non-human. In some embodiments, the subject is a human.

[0091] A suitable biological sample that can be processed in the methods disclosed herein is identified elsewhere in this disclosure or are well-known in the art and methods of analyzing such samples are within the purview of the disclosure.

[0092] Any structural elements of the devices described elsewhere in this disclosure, for example, those described under "Devices" above, are applicable to the methods disclosed herein. For example, the structures of devices and various elements of the devices as discussed elsewhere in this disclosure also applicable to the methods disclosed herein.

[0093] Certain aspects of the disclosure provide a method of enriching a sample, for example, a urine sample, for an analyte in the devices described herein. In some cases, the analyte is LAM.

[0094] Accordingly, certain aspects of the disclosure provide a method for processing a sample to enrich an analyte, the method comprising:

[0095] loading a sample in a sample chamber of a device, the sample chamber comprising a bottom comprising a pierceable element,

[0096] actuating a piercing element to pierce the pierceable element thereby causing the sample loaded into the sample chamber to flow under gravity into a reaction chamber,

[0097] incubating the sample in the reaction chamber,

[0098] inverting the device to allow the sample from the reaction chamber to flow through a column that is fluidical ly connected to the reaction chamber, wherein the column comprises a bed that binds constituents of the sample other than the analyte, and wherein the sample from the reaction chamber flows through the column under gravity to produce an analyte enriched sample.

[0099] As noted above, in some cases, instead of a sample chamber having a bottom having a pierceable element, the devices disclosed herein comprise a sample chamber having a timer channel that fluidically connects the sample chamber to the reaction chamber. An example of such timer channel is provided in FIG. 6A.

[0100] Accordingly, in certain embodiments, the disclosure provides a method of processing a sample to enrich an analyte, the method comprising:

[0101] loading a sample in a sample chamber of a device,

[0102] allowing the sample loaded into the sample chamber to flow into a reaction chamber through a timer channel,

[0103] incubating the sample in the reaction chamber,

[0104] inverting the device to allow the sample from the reaction chamber to flow through a column that is fluidically connected to the reaction chamber, wherein the column comprises a bed that binds constituents of the sample other than the analyte, and wherein the sample from the reaction chamber flows through the column under gravity to produce an analyte enriched sample.

[0105] The terms "enrich an analyte," "analyte enriched sample," and the like indicate that the sample is processed to increase the relative amount of the analyte as compared to other components in the sample. Thus, the actual concentration of the analyte in the sample may notchange and may even reduce; however, the concentration of the analyte relative to other components in the sample is increased.

[0106] The amount of a sample, such as a urine sample, added to a sample chamber can vary from 0.1 ml to 10 ml, such as about 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 ml.

[0107] After a user actuates a piercing element to pierce the pierceable element and cause the sample loaded into the sample chamber to flow to a reaction chamber, the sample can be incubated in the reaction chamber.

[0108] A sample can be incubated in the reaction chamber for an appropriate time, ranging, for example, from 1 second to 1 hour or more. For example, a urine sample can be incubated in a reaction chamber comprising a surfactant for at least or about: between 1 second and 60 seconds, such as 5 seconds, 15 seconds, 30 seconds, 45 seconds, or 60 seconds. A urine sample can be incubated in a reaction chamber comprising a surfactant for at least or about: between 1 minute and 60 minutes, such as 5 minutes, 15 minutes, 30 minutes, 45 minutes, or 60 minutes. In a specific embodiment, a urine sample can be incubated in a reaction chamber comprising a surfactant for at least 1 minute.

[0109] An appropriate incubation time can depend on the temperature of incubation. At colder temperatures, the incubation time needs to be longer as compared to incubation at warmer temperatures.

[0110] A mixture of a urine sample can be incubated in a reaction chamber comprising a surfactant at a temperature of: between 4°C and 100°C, such as 10°C, 15°C, 20°C, 25°C, 30°C, 35°C, 40°C, 50°C, 60°C, 70°C, 80°C, 90°C or 95°C.

[0111] After a suitable incubation time in a reaction chamber, a user can invert the device to allow the sample from the reaction chamber to flow through a column. The column is fluidically connected to the reaction chamber. Also, the column comprises a bed that binds constituents of the sample other than the analyte. Thus, when the sample from the reaction chamber flows through the column under gravity, it produces an analyte enriched sample, which is collected in a collection element.

[0112] In certain embodiments, the methods disclosed herein comprise allowing a treated sample to rapidly flow through the column. Thus, a sample can be allowed to flow through the column in less than 60 seconds, particularly, less than 50 seconds, such as less than 40, 30, 20, 10, or 5 seconds.

[0113] As discussed above, a collection element can be a collection chamber with or without one or more capillary channels. Accordingly, in some cases, the method comprises collecting theanalyte enriched sample in a collection element, such as a collection chamber or in one or more capillary channels of a collection chamber.

[0114] In some cases, a device disclosed herein provides a visual clue to a user that an analyte enriched sample is collected in the collection element. Thus, in certain embodiments of the methods disclosed herein, once a user observes a clue, the user proceeds with further method steps, such as removing the collection element and analyzing the collected analyte enriched sample.

[0115] In some cases, the collection element is removable from the rest of the device. Thus, in some cases, the methods comprise removing the collection element from the rest of the device.

[0116] In certain embodiments, the devices are used for pre-treatment of urine to enrich and release LAM antigen. The enriched analyte can be used in downstream detection assays, for example, lateral flow immunoassay, such as lateral flow immunoassay for LAM. Thus, in some cases, the devices disclosed here in can be used in methods to enrich LAM in a urine sample.

[0117] An analyte enriched sample collected in a collection element can be analyzed for the analyte. In certain embodiments of the methods disclosed herein, the detection of the target antigen is performed using an assay based on binding of the target antigen to a binding agent. The binding agent can be any agent that specifically binds to the target antigen. Certain nonlimiting examples of such binding agents include antibody or antigen-binding fragment thereof, specifically binding protein partners, aptamers, and the like. Additional examples of binding agents are well-known in the art and such embodiments are within the purview of the disclosure.

[0118] Any suitable binding agent-based assay can be used to detect the target antigen according to the methods disclosed herein. Certain non-limiting examples of a binding-agent based assay include lateral flow immunoassay, enzyme-linked immunosorbent assays (ELISA), radioimmunoassay (RIS), real-time immunoquantitative PCR (iqPCR), and the like. Additional examples of binding agent-based assays are well-known in the art and such embodiments are within the purview of the disclosure.EXAMPLESExample 1: An exemplary sample processing and analyte enriching device

[0119] FIGS. 1A-1B depict an exemplary sample processing and analyte enriching device 100. The device comprises a sample chamber 101 comprising a bottom comprising a pierceable element 102. The pierceable element can be a fragile foil that can be pierced with the piercing element 103 having the pointed tip 104.

[0120] The device comprises the lid 105 having the flexible element 106 that can be used to close the sample chamber.

[0121] The sample chamber is fluidically connected to the reaction chamber 108 through the pierceable element 102 and the channel 107. The reaction chamber 108 comprises one or more reagents 109.

[0122] The device further comprises the column 110. The column can comprise a bed comprising binding material, such as beads 111. The column is fluidically connected to the collection element 112, which comprises one or more capillary channels 113.

[0123] FIG. IB shows a photograph of a prototype of the exemplary device of FIG. 1A.Example 2 - An exemplary method of processing a sample using a device disclosed in Example 1

[0124] The device described in Example 1 can be used in a method of processing a sample to enrich an analyte. This method is depicted in FIGS. 2A to 2D using the device 200.

[0125] A user can load the sample 214 for example, using the pipette 215, into the sample chamber 201. The user can then close the lid 205 using the flexible element 206 so that, when closed, the lid pushes down the piercing element 203 so that the pointed tip 204 of the piercing element 203 pierces through the foil. This is shown in FIG. 2B.

[0126] When the pierceable element 202 is pierced, the sample flows down through the channel 207 and collects in the reaction chamber 208. The reaction chamber comprises one or more reagents 209. When the sample is incubated in the reaction chamber for an appropriate time under appropriate conditions, the one or more reagents act upon the sample. For example, the one or more reagents can render available an analyte in the sample. This is shown in FIG. 2C, left panel.

[0127] After appropriate incubation period, a user can invert the device 200, as shown in the middle panel of FIG. 2C so that the column 210 is now below the reaction chamber 208. In this position, the sample treated with the one or more reagents now flows under gravity through the column 210. The column comprises a bed comprising a binding material, such as beads, 211. The beads 211 specifically bind to components of the sample other than the analyte. Thus, when the sample flows through the column 211, the components of the sample otherthan the analyte are captured in the column thereby releasing the sample that is enriched in the analyte. The analyte enriched sample 216 is collected in the collection element 212. The collection element comprises one or more capillary channels 213. The capillary channels 213 absorb the analyte enriched sample. The collection element 212 is removable from the rest of the device. Thus,the collection element 212 can be removed to transfer the analyte enriched sample collected therein, which can then be analyzed as desired.Example 3: An exemplary sample processing and analyte enriching device

[0128] FIGS. 3A-3B depict an exemplary sample processing and analyte enriching device 300. The device comprises the sample chamber 301 having a bottom comprising a pierceable element 302. The sample chamber has the lid 305 comprising the flexible element 306. In this embodiment, when a user pushes down on the lid 305, the sample chamber 301 is pushed down towards the piercing element 303, which comprises piercing spikes that pierce the pierceable element 302. The reaction chamber 308 comprises one or more reagents 309. The column 310 is divided into three subunits. When the device is inverted, the sample from the reaction chamber moves under gravity through the column 301 and collects in the collection element 312. The collection element 312 can be removed from the rest of the device to transfer the analyte enriched sample collected therein, which can then be analyzed as desired.

[0129] In the exemplary device described in this Example, the column is split into multiple sections to allow ease of bead handling and improved performance. The collection element 312 allows for passive capture and metering of the final processed sample while also providing a visual indicator.Example 4: An exemplary sample processing and analyte enriching device

[0130] In the exemplary device 400 in FIG. 4A, a user can load the sample 414 for example, using the pipette 415, into the sample chamber 401. The user can push down the piercing element 403 using the lid 405 so that the pointed tip 404 of the piercing element 403 pierces through the pierceable element 402. This is shown in FIG. 4B.

[0131] When the pierceable element 402 is pierced, the sample flows down through the channel 407 and collects in the reaction chamber 408. The reaction chamber comprises one or more reagents 409. When the sample is incubated in the reaction chamber for an appropriate time under appropriate conditions, the one or more reagents act upon the sample. For example, the one or more reagents can render available an analyte in the sample. This is shown in FIG. 4B, right panel.

[0132] After appropriate incubation period, a user can invert the device 400, as shown in the middle panel of FIG. 4C so that the column 410 is now below the reaction chamber 408. In this position, the sample treated with the one or more reagents now flows under gravity through thecolumn 410. The column comprises a bed comprising a binding material, such as beads, 411. The beads 411 specifically bind to components of the sample other than the analyte. Thus, when the sample flows through the column 411, the components of the sample otherthan the analyte are captured in the column thereby releasing the sample that is enriched in the analyte. The analyte enriched sample 416 is collected in the collection element 412. The collection element 412 is removable from the rest of the device. Thus, the collection element 412 can be removed to transferthe analyte enriched sample collected therein, which can then be analyzed as desired.Example 5: An exemplary sample processing and analyte enriching device

[0133] FIGS. 5A to 5D describe a further exemplary sample processing and analyte enriching device of the disclosure. The device 500 of FIGS. 5A to 5D comprises the sample chamber 501, which is fluidical ly connected to the sample overflow chamber 502. A user can introduce the sample 514 using the pipette 515. The sample is loaded into the sample chamber 501. The user then uses the plunger 516 to push the sample into the sample overflow chamber 502. From the sample overflow chamber502, the sample trickles down through the channel 507 tothe reaction chamber 508, which comprises one or more reagents 509. This is shown in FIG. 5B, first and second panel from the left.

[0134] After appropriate incubation of the sample in the reaction chamber 508, as shown in FIGS. 5B, third and fourth panels from the left, a user can invert the device, as shown in FIG. 5C. In this orientation, the sample from the reaction chamber trickles down over the column 510. The column comprises the beads 511, which bind certain components of the treated sample. The analyte enriched sample after it passes over the column 510, is collected in the collection element 512. The collection element 512 is removal from the rest of the device. Thus, the collection element 512 can be removed to transfer the analyte enriched sample collected therein, which can then be analyzed as desired.Example 6: An exemplary sample processing and analyte enriching device

[0135] FIGS. 6A to 6E describe a further exemplary sample processing and analyte enriching device of the disclosure. The device 600 of FIG. 6A to 6E comprises the sample chamber 601, which is fluidically connected to the reaction chamber 608. A user can introduce the sample 614 using the pipette 615. The sample is loaded into the sample chamber 601. From the sample chamber 501, the sample trickles down through the channel 607 to the reaction chamber 608, which comprises one or more reagents 609. This is shown in FIGS. 6A to 6C.

[0136] After appropriate incubation of the sample in the reaction chamber 608, as shown in FIG. 6D, a user can invert the device. In this orientation, the sample from the reaction chamber 608 trickles down over the column 610. The column comprises the beads 611, which bind certain components of the treated sample. The analyte enriched sample after it passes over the column 610, is collected in the collection element 612.

[0137] The analyte enriched sample collected in the collection element 512 can be removed in variety of ways to analyze the sample as desired. Certain such embodiments are described in FIGS. 7A to 7C. For example, as shown in FIG. 7A, the analyte enriched sample can be absorbed in the absorbent swab 717 and the sample is transferred to the lateral flow assay device 718 for further analysis. This is shown in FIG. 7A.

[0138] Alternatively, as shown in FIG. 7B, the lateral flow assay device can be in fluidic contact with the analyte enriched sample to load the analyte enriched sample into the lateral flow assay device 718.

[0139] Further, the analyte enriched sample can be transferred to the lateral flow assay device 718 using the dropper 719.Example 7: An exemplary sample processing and analyte enriching device

[0140] FIGS. 8A to 8D describe a further exemplary sample processing and analyte enriching device of the disclosure. The device 800 of FIGS. 8A to 8D comprises the sample chamber 801, which is f I uid ically connected to the sample overflow chamber 816. A sample is loaded into the sample chamber 801. As shown in FIG. 8B, left panel, a predetermined amount of sample is loaded into the sample chamber 801 and any excess sample is collected in the sample overflow chamber 816. The sample trickles down through the channel 807 to the reaction chamber 808, which comprises one or more reagents 809.

[0141] Once a sample is loaded into the sample chamber 808, the device is inverted as shown in FIG. 8C, left panel. The sample is incubated in this position for a predetermined period of time, which is based on the timed dissolution time of the first dissolvable block 814. After an appropriate incubation of the sample in the reaction chamber 508, the first dissolvable block 814 dissolves and opens the fluidic connection to the column. The sample then flows through the column as shown in FIG. 8C, right panel. An analyte enriched sample is collected at the bottom of the column and further movement of the sample is blocked by the second dissolvable block 815, as shown in FIG> 8D, left panel. After a predetermined period of time, the second dissolvable block 815 dissolves and the analyte enriched sample is collected in the collectionelement 512 as shown in FIG. 8D, right panel. The collected sample can be loaded into the collection element 813, which can be separated from the rest of the device. The analyte enriched sample can then be analyzed as desired.

[0142] For reasons of completeness, various aspects of the invention are set out in the following numbered clauses:

[0143] Clause 1. A sample processing and analyte enriching device, the device comprising:

[0144] a sample chamber having a bottom comprising a pierceable element, the sample chamber fluidically connected through the pierceable element to a reaction chamber,

[0145] a piercing element that pierces the pierceable element,

[0146] a column fluidically connected to the reaction chamber, wherein the column comprises a bed that binds constituents of the sample other than the analyte thereby producing an analyte enriched sample, and

[0147] a collection element that collects the analyte enriched sample.

[0148] Clause 2. A sample processing and analyte enriching device, the device comprising:

[0149] i) in a sample loading position: a sample chamber comprising a bottom comprising a pierceable element, a piercing element that, when actuated, pierces the pierceable element thereby causing the sample loaded into the sample chamber to flow under gravity into a reaction chamber, and

[0150] ii) in an analyte enrichment position: a column fluidically connected to the reaction chamber such that the sample from the reaction chamber flows through the column under gravity, wherein the column comprises a bed that binds constituents of the sample other than the analyte thereby producing an analyte enriched sample, wherein the analyte enriched sample collects in a collection element,

[0151] wherein the device in the analyte concentration position is in a physically inverted orientation as compared to the device in the sample loading position.

[0152] Clause 3. A sample processing and analyte enriching device, the device comprising:

[0153] a sample chamber fluidically connected through a channel to a reaction chamber, wherein the channel allows a timed transfer of a sample from the sample chamber to the reaction chamber,

[0154] a column fluidically connected to the reaction chamber, wherein the column comprises a bed that binds constituents of the sample other than the analyte thereby producing an analyte enriched sample, and

[0155] a collection element that collects the analyte enriched sample.

[0156] Clause 4. The device of any one of clauses 1 to 3, wherein the sample chamber and the column are located on the same side of the reaction chamber.

[0157] Clause 5. The device of any one of clauses 1 to 4, wherein the column is located in between the reaction chamber and the collection element.

[0158] Clause 6. The device of any one of clauses 1 to 4, wherein the collection element comprises one or more capillary channels.

[0159] Clause 7. The device of clause 6, wherein the one or more capillary channels absorb the analyte enriched sample via capillary action.

[0160] Clause 8. The device of any one of the preceding clauses, where in the collection element is removable from the rest of the device.

[0161] Clause 9. The device of any one of the preceding clauses, wherein the reaction chamber comprises a surfactant.

[0162] Clause 10. The device of clause 9, wherein the surfactant increases the availability in the sample of the analyte.

[0163] Clause 11. The device of clause 10, wherein the analyte is lipoarabinomannan (LAM).

[0164] Clause 12. The device of any one of clauses 9 to 11, wherein the surfactant is a nonionic surfactant, an anionic surfactant, a cationic surfactant, an amphoteric surfactant, or a zwitterionic surfactant.

[0165] Clause 13. The device of clause 12, wherein the surfactant is the non-ionic surfactant.

[0166] Clause 14. The device of clause 13, wherein the non-ionic surfactant is polyethylene glycol hexadecyl ether, polysorbate 20, secondary alcohol ethoxylate - 9, secondary alcohol ethoxylate - 7, alcohol ethoxylate - 9, or a combination thereof.

[0167] Clause 15. The device of clause 12, wherein the surfactant is a zwitterionic surfactant.

[0168] Clause 16. The device of clause 15, wherein the zwitterionic surfactant is myristyl sulfobetaine.

[0169] Clause 17. The device of clause 12, wherein the surfactant is the ionic surfactant.

[0170] Clause 18. The device of clause 16, wherein the ionic surfactant is sodium dodecyl sulfate.

[0171] Clause 19. The device of any one of the preceding clauses, wherein the column comprises beads that specifically bind to constituents of the sample other than the analyte.

[0172] Clause 20. The device of any one of the preceding clauses, wherein the piercing element is movable between a non-piercing position and a piercing position, wherein in the non-piercing position the piercing element does not touch the pierceable element and in the piercing position the piercing element pierces the pierceable element.

[0173] Clause 21. The device of clause 20, wherein the piercing element is operably connected to a piercing actuation element that moves the piercing element from the nonpiercing position to the piercing position.

[0174] Clause 22. The device of clause 21, wherein the piercing actuation element is a lid that simultaneously closes the sample chamber and moves the piercing element from the nonpiercing position to the piercing position.

[0175] Clause 23. The device of any one of the preceding clauses, wherein the bed that binds constituents of the sample other than the analyte comprises beads.

[0176] Clause 24. The device of any one of the preceding clauses, wherein the collection element is fluid ical ly connected to an assay device that detects the analyte.

[0177] Clause 25. The device of clause 24, wherein the assay device is a lateral flow immunoassay device.

[0178] Clause 26. The device of any one of the preceding clauses, wherein the fluidic connection between the reaction chamber and the column comprises a first dissolvable block that dissolves in a predetermined time to open the fluidic connection between the reaction chamber and the column.

[0179] Clause 27. The device of any one of the preceding clauses, wherein the fluidic connection between the column and the collection element comprises a second dissolvable block that dissolves in a predetermined time to open the fluidic connection between the reaction chamber and the column.

[0180] Clause 28. A method processing a sample to enrich an analyte, the method comprising:

[0181] loading a sample in a sample chamber of a device, the sample chamber comprising a bottom comprising a pierceable element,

[0182] actuating a piercing element to pierce the pierceable element thereby causing the sample loaded into the sample chamber to flow under gravity into a reaction chamber,

[0183] incubating the sample in the reaction chamber,

[0184] inverting the device to allow the sample from the reaction chamber to flow through a column that is fluidical ly connected to the reaction chamber, wherein the column comprises a bed that binds constituents of the sample other than the analyte, and wherein the sample fromthe reaction chamber flows through the column under gravity to produce an analyte enriched sample.

[0185] Clause 29. The method of clause 28, further comprising collecting the analyte enriched sample in a collection element.

[0186] Clause 30. The method of clause 28 or 29, wherein the collection element is a collection chamber.

[0187] Clause 31. The method of clause 28 or 29, wherein the collection element comprises one or more capillary channels.

[0188] Clause 32. The method of clause 30, comprising absorbing the analyte enriched sample via capillary action in the one or more capillary channels.

[0189] Clause 33. The method of any one of clauses 28 to 32, further comprising removing the collection element from the rest of the device.

[0190] Clause 34. The method of any one of clauses 28 to 33, wherein the reaction chamber comprises a surfactant.

[0191] Clause 35. The method of clause 34, wherein the surfactant increases the availability in the sample of the analyte.

[0192] Clause 36. The method of clause 35, wherein the analyte is lipoarabinomannan (LAM).

[0193] Clause 37. The method of any one of clauses 34 to 36, wherein the surfactant is a non-ionic surfactant, an anionic surfactant, a cationic surfactant, an amphoteric surfactant, or a zwitterionic surfactant.

[0194] Clause 38. The method of clause 37, wherein the surfactant is the non-ionic surfactant.

[0195] Clause 39. The method of clause 38, wherein the non-ionic surfactant is polyethylene glycol hexadecyl ether, polysorbate 20, secondary alcohol ethoxylate - 9, secondary alcohol ethoxylate - 7, alcohol ethoxylate - 9, or a combination thereof.

[0196] Clause 40. The method of clause 37, wherein the surfactant is a zwitterionic surfactant.

[0197] Clause 41. The method of clause 40, wherein the zwitterionic surfactant is myristyl sulfobetaine.

[0198] Clause 42. The method of clause 37, wherein the surfactant is the ionic surfactant.

[0199] Clause 43. The method of clause 42, wherein the ionic surfactant is sodium dodecyl sulfate.

[0200] Clause 44. The method of any one of clauses 28 to 43, wherein the column comprises beads that specifically bind to constituents of the sample other than the analyte.

[0201] Clause 45. The method of any one of clauses 28 to 44, comprising actuating an actuation element that is operably connected to the piercing element to move the piercing element from a non-piercing position to a piercing position.

[0202] The preceding merely illustrates the principles of the present disclosure. It will be appreciated that those skilled in the art will be able to devise various arrangements which, although not explicitly described or shown herein, embody the principles of the invention and are included within its spirit and scope. Furthermore, all examples and conditional language recited herein are principally intended to aid the reader in understanding the principles of the invention and the concepts contributed by the inventors to furthering the art and are to be construed as being without limitation to such specifically recited examples and conditions. Moreover, all statements herein reciting principles, aspects, and embodiments of the invention as well as specific examples thereof, are intended to encompass both structural and functional equivalents thereof. Additionally, it is intended that such equivalents include both currently known equivalents and equivalents developed in the future, i.e., any elements developed that perform the same function, regardless of structure. The scope of the present invention, therefore, is not intended to be limited to the exemplary embodiments shown and described herein. Rather, the scope and spirit of present invention is embodied by the appended claims.

Claims

CL IMSWE CLAIM:

1. A sample processing and analyte enriching device, the device comprising: a sample chamber having a bottom comprising a pierceable element, the sample chamber fluidically connected through the pierceable element to a reaction chamber, a piercing element that pierces the pierceable element, a column fluidically connected to the reaction chamber, wherein the column comprises a bed that binds constituents of the sample other than the analyte thereby producing an analyte enriched sample, and a collection element that collects the analyte enriched sample.

2. A sample processing and analyte enriching device, the device comprising: i) in a sample loading position: a sample chamber comprising a bottom comprising a pierceable element, a piercing element that, when actuated, pierces the pierceable element thereby causing the sample loaded into the sample chamber to flow under gravity into a reaction chamber, and ii) in an analyte enrichment position: a column fluidically connected to the reaction chamber such that the sample from the reaction chamber flows through the column under gravity, wherein the column comprises a bed that binds constituents of the sample other than the analyte thereby producing an analyte enriched sample, wherein the analyte enriched sample collects in a collection element, wherein the device in the analyte concentration position is in a physically inverted orientation as compared to the device in the sample loading position.

3. A sample processing and analyte enriching device, the device comprising: a sample chamber fluidically connected through a channel to a reaction chamber, wherein the channel allows a timed transfer of a sample from the sample chamber to the reaction chamber, a column fluidically connected to the reaction chamber, wherein the column comprises a bed that binds constituents of the sample other than the analyte thereby producing an analyte enriched sample, anda collection element that collects the analyte enriched sample.

4. The device of any one of claims 1 to 3, wherein the sample chamber and the column are located on the same side of the reaction chamber.

5. The device of any one of claims 1 to 4, wherein the column is located in between the reaction chamber and the collection element.

6. The device of any one of claims 1 to 4, wherein the collection element comprises one or more capillary channels.

7. The device of claim 6, wherein the one or more capillary channels absorb the analyte enriched sample via capillary action.

8. The device of any one of the preceding claims, where in the collection element is removable from the rest of the device.

9. The device of any one of the preceding claims, wherein the reaction chamber comprises a surfactant.

10. The device of claim 9, wherein the surfactant increases the availability in the sample of the analyte.

11. The device of claim 10, wherein the analyte is lipoarabinomannan (LAM).

12. The device of any one of claims 9 to 11, wherein the surfactant is a non-ionic surfactant, an anionic surfactant, a cationic surfactant, an amphoteric surfactant, or a zwitterionic surfactant.

13. The device of claim 12, wherein the surfactant is the non-ionic surfactant.

14. The device of claim 13, wherein the non-ionic surfactant is polyethylene glycol hexadecyl ether, polysorbate 20, secondary alcohol ethoxylate -9, secondary alcohol ethoxylate - 7, alcohol ethoxylate - 9, or a combination thereof.

15. The device of claim 12, wherein the surfactant is a zwitterionic surfactant.

16. The device of claim 15, wherein the zwitterionic surfactant is myristyl sulfobetaine.

17. The device of claim 12, wherein the surfactant is the ionic surfactant.

18. The device of claim 16, wherein the ionic surfactant is sodium dodecyl sulfate.

19. The device of any one of the preceding claims, wherein the column comprises beads that specifically bind to constituents of the sample other than the analyte.

20. The device of any one of the preceding claims, wherein the piercing element is movable between a non-piercing position and a piercing position, wherein in the non-piercing position the piercing element does not touch the pierceable element and in the piercing position the piercing element pierces the pierceable element.

21. The device of claim 20, wherein the piercing element is operably connected to a piercing actuation element that moves the piercing element from the non-piercing position to the piercing position.

22. The device of claim 21, wherein the piercing actuation element is a lid that simultaneously closes the sample chamber and moves the piercing element from the nonpiercing position to the piercing position.

23. The device of any one of the preceding claims, wherein the bed that binds constituents of the sample other than the analyte comprises beads.

24. The device of any one of the preceding claims, wherein the collection element is fluidically connected to an assay device that detects the analyte.

25. The device of claim 24, wherein the assay device is a lateral flow immunoassay device.

26. The device of any one of the preceding claims, wherein the fluidic connection between the reaction chamber and the column comprises a first dissolvable block that dissolves in a predetermined time to open the fluidic connection between the reaction chamber and the column.

27. The device of any one of the preceding claims, wherein the fluidic connection between the column and the collection element comprises a second dissolvable block that dissolves in a predetermined time to open the fluidic connection between the reaction chamber and the column.

28. A method processing a sample to enrich an analyte, the method comprising: loading a sample in a sample chamber of a device, the sample chamber comprising a bottom comprising a pierceable element, actuating a piercing element to pierce the pierceable element thereby causing the sample loaded into the sample chamber to flow under gravity into a reaction chamber, incubating the sample in the reaction chamber, inverting the device to allow the sample from the reaction chamber to flow through a column that is fluidically connected to the reaction chamber, wherein the column comprises a bed that binds constituents of the sample other than the analyte, and wherein the sample from the reaction chamber flows through the column under gravity to produce an analyte enriched sample.

29. The method of claim 28, further comprising collecting the analyte enriched sample in a collection element.

30. The method of claim 28 or 29, wherein the collection element is a collection chamber.

31. The method of claim 28 or 29, wherein the collection element comprises one or more capillary channels.

32. The method of claim 30, comprising absorbing the analyte enriched sample via capillary action in the one or more capillary channels.

33. The method of any one of claims 28 to 32, further comprising removing the collection element from the rest of the device.

34. The method of any one of claims 28 to 33, wherein the reaction chamber comprises a surfactant.

35. The method of claim 34, wherein the surfactant increases the availability in the sample of the analyte.

36. The method of claim 35, wherein the analyte is lipoarabinomannan (LAM).

37. The method of any one of claims 34 to 36, wherein the surfactant is a non-ionic surfactant, an anionic surfactant, a cationic surfactant, an amphoteric surfactant, or a zwitterionic surfactant.

38. The method of claim 37, wherein the surfactant is the non-ionic surfactant.

39. The method of claim 38, wherein the non-ionic surfactant is polyethylene glycol hexadecyl ether, polysorbate 20, secondary alcohol ethoxylate -9, secondary alcohol ethoxylate - 7, alcohol ethoxylate - 9, or a combination thereof.

40. The method of claim 37, wherein the surfactant is a zwitterionic surfactant.-SO-41. The method of claim 40, wherein the zwitterionic surfactant is myristyl sulfobetaine.

42. The method of claim 37, wherein the surfactant is the ionic surfactant.

43. The method of claim 42, wherein the ionic surfactant is sodium dodecyl sulfate.

44. The method of any one of claims 28 to 43, wherein the column comprises beads that specifically bind to constituents of the sample other than the analyte.

45. The method of any one of claims 28 to 44, comprising actuating an actuation element that is operably connected to the piercing element to move the piercing element from a non-piercing position to a piercing position.

Images