Systems and methods for digitized, multiplexed, extracellular vesicle-derived biomarker diagnostic chip laboratories

Abstract

In one embodiment, a method for separating and detecting one or more biomarkers of interest on a dielectrophoresis device includes receiving a first biological sample containing one or more biomarkers of interest onto a dielectrophoresis (DEP) electrode array, applying a DEP force through specific electrodes of the DEP electrode array, and separating and detecting one or more biomarkers of interest on the DEP electrode array. Wherein the intensity, direction, and time period of the DEP force are directed to one or more biomarkers of interest of the first biological sample, and determining an amount of the one or more biomarkers of interest of the first biological sample via a digital sensor.

Description

Technical Field

[0001] The present disclosure relates generally to lab-on-a-chip diagnostic platforms, and in particular to detecting extracellular vesicle biomarkers using lab-on-a-chip diagnostics. Background Art

[0002] There is a growing demand for diagnostic markers for early disease detection, which can increase the chances of survival. Early disease detection requires sensitive diagnostic tools to detect small amounts of biomarkers that indicate disease. Few methods have the sensitivity and specificity required to detect diseases such as cancer and neurodegenerative diseases before considerable progression occurs. Therefore, it may be useful to develop highly sensitive lab-on-a-chip tests for early disease detection.

[0003] Extracellular vesicles (EVs) are membranous nanoparticles that facilitate intercellular communication systems via their biomolecular components (e.g., proteins, lipids, carbohydrates, and nucleic acids). EVs are dense information compartments that are continuously released from the original cells and contain biomarkers that mimic their original cells. EVs are present in biological fluids (e.g., blood, urine, cerebrospinal fluid, etc.), and EV-associated markers have a longer half-life and higher stability than free circulating biomarkers. Therefore, EVs provide an accessible source of biomarkers that are continuously released from living cells in vivo.

[0004] Detection methods for cancer and neurodegenerative diseases rely on expensive, time-consuming and often invasive methods (e.g., tissue biopsy, computed tomography, magnetic resonance imaging, endoscopy, etc.), many of which have no tests available for early detection. Currently, liquid biopsy tests rely on free circulating markers released during tumor cell death, rather than ongoing cellular processes such as EV secretion. Therefore, EVs represent a valuable biological compartment that can be used to detect disease-associated biomarkers early and minimally invasively from their parent cells (e.g., tumor cells, neurons affected by neurodegeneration, inflammatory cells, etc.). Existing EV-based detection methods either use infected "strong" concentration methods such as centrifugation and size exclusion, or rely on dilute circulating concentrations. Many existing EV marker test methods use fluorescence-based detection, which has limited sensitivity and specificity. Therefore, it may be useful to improve existing technologies by using electronic methods for effective biomarker concentration and sensitive, specific biomarker detection. Summary of the invention

[0005] In certain embodiments, a biological sample can be loaded onto a solid-state detection device, where DEP can be used to concentrate and / or separate EVs 102. The detection device (e.g., a lab-on-a-chip device) can utilize integrated microfluidics (conventional / pneumatic and / or digital) to partition the concentrated EV volume and automate multi-marker antibody-based capture. As an example and not limitation, a sensor can be used to directly quantify the corresponding biomarker concentration, where the data from the sensor can be processed by one or more algorithms to produce a comprehensive result.

[0006] In certain embodiments, a method for separating and detecting one or more biomarkers of interest on a dielectrophoresis (DEP) device may include receiving a first biological sample containing one or more biomarkers of interest 104 onto a DEP electrode array of the DEP device. In certain embodiments, one or more processors of the DEP device may instruct the DEP device to apply a DEP force through one or more electrodes of the DEP electrode array. As an example and not limitation, one or more processors of the DEP device may instruct the DEP device to apply a DEP force of a specific intensity, direction, time period, or other configurable setting specifically for one or more biomarkers of interest (e.g., EV) of the biological sample. In certain embodiments, one or more sensors of the DEP device may determine the amount of one or more target biomarkers of interest of the biological sample.

[0007] In certain embodiments, a method for detecting a biomarker of interest on a DEP device may include receiving a first biological sample containing one or more biomarkers of interest onto one or more arrays of a DEP device. In certain embodiments, a current may be applied through one or more electrode arrays to separate the one or more biomarkers from a plurality of biomarkers in the biological sample. In certain embodiments, the DEP device may filter a specific biomarker of interest from a plurality of biomarkers in the biological sample into one or more chambers of the DEP device via one or more microfluidics. In certain embodiments, one or more specific biomarkers may be labeled with one or more markers and then detected via one or more sensors, where the amount of each biomarker may be determined and displayed.

[0008] In certain embodiments, a method for detecting at least one biomarker of interest on a device may include receiving a first biological sample and transferring the first biological sample to a first chamber via a microfluidic channel. As an example and not limitation, a first treatment may be performed on the first biological sample in the first chamber. In certain embodiments, the first biological sample may be transferred from the first chamber to the second chamber via a microfluidic channel. As an example and not limitation, a second treatment may be performed on the first biological sample in the second chamber. In certain embodiments, the first biological sample may be transferred from the second chamber to the third chamber via a microfluidic channel. As an example and not limitation, a third treatment may be performed on the first biological sample in the third chamber.

[0009] In certain embodiments, a method for separating at least one biomarker of interest on a solid-state device may include receiving a first biological sample on a solid-state device. The solid-state device may then transfer the first biological sample to a DEP chamber. As an example and not limitation, one or more processes may be performed on the first biological sample in the DEP chamber. In certain embodiments, the solid-state device may transfer the first biological sample through a microfluidic (MF) channel to a labeling chamber, where one or more biomarkers of interest of the first biological sample may be labeled based on one or more features.

[0010] In certain embodiments, the solid-state device can transfer one or more biomarkers of interest from the labeling chamber to the sensor chamber. In certain embodiments, the solid-state device can digitally determine the amount of the one or more biomarkers of interest in the sensor chamber based on the amount of labeling within the first biological sample.

[0011] In certain embodiments, a method 1800 for separating at least one biomarker of interest on a solid-state device. In step 1810, one or more processors of the solid-state device may provide instructions to receive a first biological sample into the solid-state device. As an example and not limitation, the first biological sample may include one or more biomarkers of interest. As another example and not limitation, the first biological sample may be input via a microfluidic (MF) channel.

[0012] In certain embodiments, one or more processors of the solid-state device may provide instructions to transfer the first biological sample to the DEP chamber via MF and apply an AC current to the first biological sample through one or more electrodes of the DEP chamber.

[0013] In certain embodiments, one or more processors of the solid-state device may provide instructions to one or more components of the solid-state device to transfer the first biological sample from the DEP chamber to the labeling chamber from the MF.

[0014] In a particular embodiment, one or more processors of the solid-state device may provide instructions to one or more components of the solid-state device to assign a tag to each of one or more biomarkers of interest of the first biological sample. As an example and not limitation, each tag may define a specific label among a plurality of labels. In a particular embodiment, a first tag may be assigned to a first biomarker of interest of the first biological sample based on one or more specific features. In a particular embodiment, a second tag may be assigned to a second biomarker of interest of the first biological sample based on one or more specific features.

[0015] In certain embodiments, one or more processors of the solid-state device may provide instructions to one or more components of the solid-state device to transfer a first biological sample to one or more detector chambers via MF, wherein each detector chamber is programmed for a specific biomarker of interest.

[0016] In particular embodiments, one or more processors of the solid-state device may provide instructions to one or more components of the solid-state device to digitally determine the amount of one or more biomarkers of interest based on the assigned labels via one or more detectors within one or more detector chambers.

[0017] There are certain technical challenges to early cancer detection. One technical challenge may include reliance on free circulating markers released during tumor cell death. A solution to this challenge proposed by embodiments disclosed herein may be a solid-state device for isolating specific biomarker concentrations. Another technical challenge may include detecting biomarker concentrations. A solution to this challenge proposed by embodiments disclosed herein may be a solid-state device for isolating specific biomarkers, but also digitally quantifying biomarker concentrations.

[0018] Certain embodiments disclosed herein may provide one or more technical advantages. A technical advantage of an embodiment may include utilizing dielectrophoresis (DEP) on a solid-state device. Another technical advantage of an embodiment may include utilizing multiple chambers on a solid-state device, each of which may perform a specific treatment on a biological sample. Certain embodiments disclosed herein may not provide, provide some or all of the above technical advantages. In view of the drawings, description, and claims of the present disclosure, one or more other technical advantages may be apparent to those skilled in the art.

[0019] The embodiments disclosed herein are only examples, and the scope of the present disclosure is not limited to them. Specific embodiments may include all or some of the following or exclude the following: components, elements, features, functions, operations or steps of the embodiments disclosed herein. The embodiments according to the present invention are particularly disclosed in the appended claims relating to methods, devices or systems, wherein any feature mentioned in one claim category, such as a method, can also be claimed in another claim category, such as a system. The dependencies or references returned to the appended claims are selected only for formal reasons. However, any subject matter caused by intentionally returning to any previous claim (especially multiple dependencies) can also be claimed, so that any combination of claims and their features can be disclosed and claimed, regardless of the dependencies selected in the appended claims. The subject matter that can be claimed includes not only the combination of features as set forth in the appended claims, but also any other combination of features in the claims, wherein each feature mentioned in the claims can be combined with any other feature in the claims or a combination of other features. In addition, any of the embodiments and features described or depicted herein can be claimed in a separate claim and / or in any combination with any embodiment or feature described or depicted herein or any of the features of the appended claims. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] Figure 1A Example diagrams of exemplary surface chemistries and capture methods are shown.

[0021] Figure 1B Example diagrams of exemplary surface chemistries and capture methods are shown.

[0022] Figure 1C Example diagrams of exemplary surface chemistries and capture methods are shown.

[0023] Figure 1D Example diagrams of exemplary surface chemistries and capture methods are shown.

[0024] Figure 1E Example diagrams of exemplary surface chemistries and capture methods are shown.

[0025] Figure 2 An example flow diagram of sample preparation and separation and / or concentration chambers is shown.

[0026] Figure 3 An example diagram showing a cross-sectional view of the dielectrophoresis concept.

[0027] Figure 4 An example diagram of a dielectrophoresis electrode array is shown.

[0028] Figure 5AAn example isometric diagram of a generalized fluidic delivery method is shown.

[0029] Figure 5B An example isometric diagram of a generalized fluidic delivery method is shown.

[0030] Figure 6 An example diagram showing the device architecture for interrogating multiple EV-derived biomarkers from one or more biological samples using fluidic multiplexing.

[0031] Figure 7 An example diagram showing the device architecture for interrogating multiple EV-derived biomarkers from one or more biological samples using fluidic multiplexing.

[0032] Figure 8 An example diagram showing the device architecture for interrogating multiple EV-derived biomarkers from one or more biological samples using fluidic multiplexing.

[0033] Fig. 9 An example schematic diagram showing the open circuit potential of a working electrode is shown.

[0034] Fig.10 An example schematic diagram showing an exemplary electrode configuration is shown.

[0035] Fig.11 An example diagram showing an electrode configuration in which an embodiment may operate.

[0036] Fig.12 An example schematic diagram showing an exemplary electrode configuration is shown.

[0037] Fig.13 An example diagram showing an electrode configuration in which an embodiment may operate.

[0038] Fig.14 An example method of separating and detecting biomarkers of interest via a dielectrophoresis device is shown.

[0039] Fig.15 An example method for detecting a biomarker of interest on a dielectrophoresis device is shown.

[0040] Fig.16 Example methods for detecting at least one biomarker of interest on a device are shown.

[0041] Fig.17 An example method for separating at least one biomarker of interest on a solid-state device is shown.

[0042] Fig.18 An example method for separating at least one biomarker of interest on a solid-state device is shown.

[0043] Fig.19 A diagram of an exemplary computer system is shown. DETAILED DESCRIPTION

[0044] Figure 1A An example diagram 100 of an exemplary surface chemistry and capture method is shown. As used herein, "extracellular vesicle (EV)" may refer to a membranous nanoparticle secreted from a cell that can facilitate intercellular communication via its biomolecular components. In certain embodiments, one or more EVs 102 may contain a biomarker 104 on the surface of the EV 102. The previously discussed properties of the EV 102 may provide an opportunity for early detection of biomarkers corresponding to early-stage diseases. Therefore, combined detection of the presence of multiple cancer-related biomarkers from EV 102, as well as analysis with advanced machine learning (ML) algorithms, can facilitate sensitive and specific diagnosis of early-stage cancer and other diseases from biological fluids. Disclosed herein are compositions, methods, and exosome detection devices for biomarker detection.

[0045] In certain embodiments, a biological sample can be loaded onto an electrode array, where dielectrophoresis (DEP) can be used to concentrate and / or separate EVs 102. A detection device (e.g., a lab-on-a-chip device) can utilize integrated microfluidics (conventional / pneumatic and / or digital) to partition the concentrated EV volume and automate multi-marker antibody-based capture. As an example and not limitation, an electrochemical sensor can be used to directly quantify the corresponding biomarker concentration, where the data from the sensor can be processed by one or more algorithms to produce a comprehensive result.

[0046] In certain embodiments, the biological fluid may be collected via standard point of care procedures, including but not limited to vacuum tube-based blood draws (e.g., for blood), spinal taps (e.g., for cerebrospinal fluid), and urine collection devices (e.g., for urine). In certain embodiments, the biological fluid may be processed to remove interfering cells. As an example and not limitation, processing of the biological fluid may involve centrifugation, membrane-based filtration, and / or other standard preparation procedures to prepare the biological sample for testing. The resulting sample may be plasma, serum, CSF, and / or urine.

[0047] In certain embodiments, a prepared biological sample may flow through an array of powered electrodes that apply dielectrophoretic (DEP) forces to particles within the biological sample. The strength and direction of the DEP forces may be specific to a variety of biological particles within the sample, thereby allowing EVs 102 to be separated and further concentrated from the biological sample. As used herein, "dielectrophoresis (DEP)" may refer to the phenomenon of forces being applied to dielectric particles, molecules, or macromolecular structures in aqueous or organic solutions when they are subjected to a non-uniform electric field. In certain embodiments, the time period for which the DEP forces are applied may be specific to a particular biomarker of interest within the biological sample.

[0048] In certain embodiments, the isolated and concentrated EVs may be mixed with immunochemical reagents and allowed to incubate. Following immunochemistry, the corresponding sample or samples may be loaded onto a sensor array. As an example and not limitation, the original volume of EV 102 may or may not be split into multiple droplets at any point during this process, depending on the necessary workflow. In certain embodiments, the sensor array may contain one or more digital sensors, each of which can determine the amount of one or more biomarkers of interest (e.g., EV 102).

[0049] In certain embodiments, one or more EVs 102 may be floating in a solution, wherein the biomarker 104 may be located on the surface of the EV 102. By way of example and not limitation, the labeling of the biomarker 104 may include, but is not limited to, binding of an antibody, an antibody conjugated to an enzyme, an antibody conjugated to a metal nanoparticle, an antibody conjugated to a cleavable single-stranded DNA barcode, an antibody conjugated to a metal nanoparticle and conjugated to a cleavable single-stranded DNA barcode, an antibody conjugated to an enzyme and conjugated to a cleavable single-stranded DNA barcode, an antibody conjugated to an enzyme and conjugated to a cleavable single-stranded DNA barcode, and / or an antibody conjugated to an enzyme and conjugated to a cleavable single-stranded DNA barcode. Cleavable linkages to the antibody may include, but are not limited to, proteolytic, chemical, electrochemical, and photolytically labile compounds. By way of example and not limitation, the EV 102 may contain a specific biomarker of interest on the surface of the EV 102, wherein a detector surface may be functionalized to capture the EV 102 as well as the biomarker 104, wherein the detectable event may be used to quantify the presence of the biomarker. Table 1 below summarizes the various configurations, EV tags, tag release mechanisms, surface capture mechanisms, and detected events.

[0050] In certain embodiments, the biomarkers 104 can be labeled with one or more tags and / or markers.

[0051] Table 1: Exemplary configurations of EV tags, tag release mechanisms, surface capture mechanisms, and detected events

[0052]

[0053]

[0054] As shown in Table 1, "ssDNA" may refer to single-stranded DNA, and "MNP" may refer to metal nanoparticles. Figure 1A As shown, graph 100 represents configuration "A", as shown in Table 1, where there is no EV tag or tag release mechanism, the surface capture mechanism is a biomarker-specific antibody, and the event detected by binding event 110 is antibody-biomarker binding.

[0055] In certain embodiments, a binding event 110 can be detected by a sensor between a biomarker 104 and a capture molecule 120 of an EV 102. By way of example and not limitation, the capture molecule 120 can be captured by an antibody. As used herein, a "binding event" 110 can refer to a detectable event used to quantify the presence of a particular biomarker 104. By way of example and not limitation, when a binding event 110 occurs, the observed event can be a change in the capacitance of the surface chemistry 122 or an increase in the number of charge carriers, both of which can be measured electronically. Because the sensitivity of a particular biomarker 104 can vary, the detection device can be adjusted to a particular range of interest.

[0056] like Figure 1A As shown in FIG. 100 , the capture molecule 120 may be an antibody. As an example and not limitation, a method of capturing an EV-derived biomarker (e.g., biomarker 104) and / or an EV-derived biomarker tag may include exposing an electrode surface 124 connected to an antibody 120 (e.g., a capture molecule) directly to an EV 102 having the biomarker 104, such that the biomarker 104 attached to the EV 102 directly binds to the antibody 120 and is captured by it.

[0057] In certain embodiments, methods of capturing EV-derived biomarkers and / or EV-derived biomarker tags may include utilizing the chemical reactivity of surface chemistry 122 to covalently attach antibodies to a surface, wherein the antibodies 120 may directly bind to their EV-derived biomarkers 104 .

[0058] In a particular embodiment, one or more surfaces 124 of the field effect transistor can be functionalized using standard surface chemical treatments so that chemical functional groups can be physically and / or chemically bound to metals, metal oxides, glassy carbon, graphene, graphene nanoribbons, carbon nanotubes, semiconductors, and / or dielectric surfaces through covalent bonds. As used herein, "surface chemistry" 122 may refer to physical and chemical phenomena occurring at the interface between two phases. Surface chemistry 122 may include molecules having multiple chemical and / or physical reaction sites, wherein one site may interact physically and / or chemically with the surface, and another site is used to directly conjugate nucleic acids, proteins, and / or other molecules of interest. Examples of surface chemistry 122 may include, but are not limited to, molecules, nanoparticles, and / or biomolecules containing functional groups such as silanes, thiols, disulfides, phosphonates, phosphonic acids, diazos, olefins, carboxylic acids, alkynes, alkanes, amines, ketones, esters, aldehydes, alcohols, amides, imines, hydrazines, ethers, nitriles, aromatic hydrocarbons, halides, and azides. By way of example and not limitation, the surface chemistry 122 can be directly conjugated to nucleic acids, proteins, glycans, lipids, and / or other molecules used in capturing and / or detecting biomarkers of interest. In certain embodiments, the insulated gate field effect transistor (IGFET), extended gate field effect transistor (EGFET), and / or ion sensitive field effect transistor (ISFET) structures can be functionalized with a gate oxide instead of a metal gate.

[0059] In certain embodiments, methods of making one or more electrodes for detecting one or more EV-derived biomarkers may include functionalizing a metal and / or metal oxide surface (hereinafter referred to as a "surface") with one or more chemical groups, and may be referred to herein as surface chemistry 122. As an example and not limitation, the chemical reactivity of the surface may be utilized to covalently attach single-stranded DNA to the surface. As another example and not limitation, the intrinsic surface reactivity may be utilized to directly attach the functionalized single-stranded DNA to the surface.

[0060] In certain embodiments, the working electrode surface 124 can function as a detector for one or more biomarkers 104. As used herein, "working electrode surface" can refer to one or more electrodes in an electrochemical system on which a reaction of interest occurs. In certain embodiments, one or more EVs 102 can then be brought into close proximity with a metal gate of one or more working electrode surfaces 124 during capture and / or detection of the biomarker 104 mediated by biomolecules and / or molecules conjugated to the gate surface (e.g., DNA-based hybridization, antibody binding, and / or enzymatic reactions). As an example and not limitation, one or more EVs 102 can be concentrated by DEP and brought into close proximity with a metal gate (e.g., capture molecules 120) of one or more working electrode surfaces 124. In certain embodiments, one or more EVs 102 within a biological sample can be labeled via one or more methods, wherein the labeled EVs can be attached to the surface of the working electrode surface 124. As an example and not limitation, one or more EVs 102 and / or biomarkers 104 can be labeled with one or more labels corresponding to one or more specific features of the EVs 102 and / or biomarkers 104. As another example and not limitation, one or more EVs 102 and / or biomarkers 104 may be labeled with one or more specific markers, where each marker may correspond to a specific feature of the EVs 102 and / or biomarkers 104. It is understood that the terms labeling, tagging, and marking may be used interchangeably to describe the process of identifying one or more specific features of one or more EVs 102 and / or biomarkers 104. Although the present disclosure discusses labeling, tagging, and / or marking specific features of one or more EVs 102 and / or biomarkers 104, the present disclosure contemplates any suitable method for identifying and characterizing the one or more specific features. In a particular embodiment, the tags on one or more EVs 102 may be cleaved from the corresponding biomarkers 104, where the working electrode surface 124 may capture only the tags at the surface and not the EVs 102 via surface chemistry 122. As an example and not limitation, methods of releasing tags specific to EV-derived biomarkers may include linkers that can be cleaved chemically, photolytically, and / or electrochemically. The linker between the biomarker 104 capable of chemical cleavage and the detection biomolecule and / or molecule may include, but is not limited to, esters, carbamates, dialkoxydiphenylsilane azo, diazo, acylhydrazones, nitrobenzenesulfonamides, acylsulfonamides, and / or disulfides. The linker between the biomarker 104 capable of photolytic cleavage and the detection biomolecule and / or molecule may include, but is not limited to, nitrophenyl ethyl ether and / or benzoyl ester. The linker between the biomarker 104 capable of electrochemical cleavage and the detection biomolecule and / or molecule may include, but is not limited to, aryl esters and imines.

[0061] In certain embodiments, digital sensing may occur when an EV biomarker of interest 104 is detected by a change in ion concentration near the gate surface of the working electrode surface 124, mediated by, for example, an antibody binding event, a nucleic acid hybridization event, and / or an enzymatic reaction. As an example and not limitation, the interaction between the biomarker 104 and its capture antibody 120 on the conjugated gate surface modulates the electrical properties of the gate of the field effect transistor, resulting in a change in the voltage or current across the transistor, depending on the surrounding circuit configuration. This electrical signal can then be amplified and digitized using an analog-to-digital converter.

[0062] In certain embodiments, when a force (eg, DEP force, AC current) is applied to a biological sample within one or more chambers of a solid-state DEP device, the applied force can cause a change in the ion concentration of the biological sample.

[0063] In certain embodiments, the digital output of one or more biomarkers 104 may be analyzed by one or more machine learning algorithms. As an example and not limitation, the one or more machine learning algorithms may include supervised learning algorithms, unsupervised learning algorithms, semi-supervised learning algorithms, deep learning algorithms, and / or reinforcement learning algorithms. In certain embodiments, the deep learning algorithm may include any artificial neural network (ANN) that can be used to learn deep representations and abstractions from a large amount of data. For example, the deep learning algorithm may include ANNs such as multilayer perceptrons (MLPs), autoencoders (AEs), convolutional neural networks (CNNs), recurrent neural networks (RNNs), long short-term memory (LSTMs), gated recurrent units (GRUs), restricted Boltzmann machines (RBMs), deep belief networks (DBNs), bidirectional recurrent deep neural networks (BRDNNs), generative adversarial networks (GANs) and deep Q networks, neural autoregressive distribution estimation (NADEs), adversarial networks (ANs), attention models (AMs), deep reinforcement learning, etc.

[0064] In certain embodiments, the digitized output of one or more biomarkers 104 may be analyzed by one or more classification machine learning algorithms or functions, which may include any algorithm that can utilize a supervised learning model (e.g., logistic regression, naive Bayes, stochastic gradient descent (SGD), k-nearest neighbors, decision trees, random forests, support vector machines (SVM), etc.) to learn from the data input to the supervised learning model and make new observations or classifications based thereon.

[0065] Although the present disclosure refers to the above machine learning algorithms, the present disclosure contemplates any suitable machine learning algorithms. In certain embodiments, one or more machine learning algorithms can analyze a variety of data, thereby allowing detection of early stage cancer (e.g., stage I, stage II) and / or other diseases of interest.

[0066] Figure 1B An example diagram 130 of an exemplary surface chemistry 122 and capture method is shown. In certain embodiments, the EV 102 may contain one or more biomarkers 104 on the surface of the EV 102. In certain embodiments, the tag 140 may include, for example, an antibody, wherein the tag 140 may bind to the biomarker 104 on the EV 102, and wherein the tag 140 may be captured by the antibody 120 on the surface. As another example and not limitation, the tag 140 may include an antibody and an MNP, wherein the capture antibody 120 may bind to the tag antibody, thereby causing a MNP proximity event. In certain embodiments, as Figure 1B As shown, Figure 130 may represent configuration "B" of Table 1, wherein label 140 may indicate an antibody, there is no label release mechanism, the surface capture mechanism is binding of a second (capture) antibody to the EV tag antibody, and the event detected at binding event 110 is antibody-antibody binding.

[0067] In a specific embodiment, Figure 1B As shown, graph 130 may represent configuration "D" of Table 1. As an example and not limitation, tag 140 may indicate antibody-MNP without a tag release mechanism, the surface capture mechanism may be binding of a second capture antibody to EV tag antibody 140, and the event detected at binding event 110 may be a MNP proximity event.

[0068] In particular embodiments, event 110 may be detected via working electrode surface 124 and surface chemistry 122 .

[0069] Figure 1C An example diagram 150 of an exemplary surface chemistry 122 and capture method is shown. In certain embodiments, the EV 102 may contain one or more biomarkers 104 and one or more tags 140 on the surface of the EV 102. In certain embodiments, as Figure 1C As shown, graph 150 may represent configuration "C" of Table 1, wherein tag 140 may include an antibody-enzyme conjugate without a tag release mechanism, the surface capture mechanism may be binding of a secondary antibody 120 to an EV tag antibody, and the event detected at event 160 may be enzyme activity.

[0070] In particular embodiments, event 160 may be detected via working electrode surface 124 and surface chemistry 122 .

[0071] Figure 1D An example diagram 170 of an exemplary surface chemistry 122 and capture method is shown. In certain embodiments, the EV 102 may contain one or more biomarkers 104 and one or more tags 140 on the surface of the EV 102. In certain embodiments, as Figure 1D As shown, diagram 170 can represent configuration "E-1" of Table 1, where the tag 140 can be an antibody-ssDNA barcode with a chemical tag release mechanism. As an example and not limitation, the tag release mechanism can be represented by step 180, where the tag can be cleaved from the EV 102. In this example, the biomarker 104 and the tag 140 can be measured in the absence of the EV 102. As an example and not limitation, the tag 140 can represent an antibody-DNA and / or antibody-DNA-MNP combination. In the example of configuration "E-1", the surface capture mechanism can be a DNA barcode hybridized with a complementary ssDNA, where the detected binding event 110 can be a DNA hybridization.

[0072] In a specific embodiment, Figure 1D As shown, the diagram 170 can represent the configuration "E-2" of Table 1, wherein the label 140 can be an antibody-ssDNA barcode with a photo label release mechanism. In the example of configuration "E-2", the surface capture mechanism can be hybridization of the DNA barcode with complementary ssDNA, wherein the detected binding event 110 can be a DNA hybridization.

[0073] In a specific embodiment, Figure 1D As shown, the diagram 170 can represent the configuration "E-3" of Table 1, where the tag 140 can be an antibody-ssDNA barcode with an electrochemical tag release mechanism. In the example of configuration "E-3", the surface capture mechanism can be hybridization of the DNA barcode with complementary ssDNA, where the detected binding event 110 can be a DNA hybridization.

[0074] In a particular embodiment, the diagram 170 can represent the configuration "G-1" of Table 1, where the tag 140 can be an antibody-ssDNA barcode-MNP with a chemical tag release mechanism. In the example of configuration "G-1", the surface capture mechanism can be hybridization of the DNA barcode to the complementary ssDNA, where the detected binding event 110 can be the proximity of the MNP.

[0075] In a particular embodiment, the diagram 170 can represent the configuration "G-2" of Table 1, where the tag 140 can be an antibody-ssDNA barcode-MNP with a photosensitive tag release mechanism. In the example of configuration "G-2", the surface capture mechanism can be hybridization of the DNA barcode with complementary ssDNA, where the detected binding event 110 can be the proximity of the MNP.

[0076] In a particular embodiment, the diagram 170 can represent the configuration "G-3" of Table 1, where the tag 140 can be an antibody-ssDNA barcode-MNP with an electrochemical tag release mechanism. In the example of configuration "G-3", the surface capture mechanism can be hybridization of the DNA barcode to the complementary ssDNA, where the detected binding event 110 can be the proximity of the MNP.

[0077] In particular embodiments, event 110 may be detected via working electrode surface 124 and surface chemistry 122 .

[0078] Figure 1E An example diagram 190 of an exemplary surface chemistry 122 and capture method is shown. In certain embodiments, the EV 102 may contain one or more biomarkers 104 and one or more tags 140 on the surface of the EV 102. In certain embodiments, as Figure 1E As shown, Figure 190 can represent configuration "F-1" of Table 1, where the EV tag can be an antibody-ssDNA barcode with a chemical tag release mechanism. As an example and not limitation, the tag release mechanism can be represented by step 180, where the tag can be cleaved from the EV 102. In this example, the tag 140 can be measured in the absence of the EV 102. As an example and not limitation, the tag 140 can represent an antibody-DNA combination and / or an antibody-DNA-enzyme combination. In the example of configuration "F-1", the surface capture mechanism can be a DNA barcode hybridized with a complementary ssDNA, where the detected event 160 can be a polymerase activity using a DNA polymerase.

[0079] In a particular embodiment, the diagram 190 can represent the configuration "F-2" of Table 1, where the tag 140 can be an antibody-ssDNA barcode with a photosensitive tag release mechanism. In the example of configuration "F-2", the surface capture mechanism can be hybridization of the DNA barcode with complementary ssDNA, where the detected event 160 can be polymerase activity using a DNA polymerase.

[0080] In a particular embodiment, the diagram 190 can represent configuration "F-3" of Table 1, where the tag 140 can be an antibody-ssDNA barcode with electrochemical tag release. In the example of configuration "F-3", the surface capture mechanism can be hybridization of the DNA barcode to the complementary ssDNA, where the detected event 160 can be polymerase activity using a DNA polymerase.

[0081] In a particular embodiment, the diagram 190 can represent the configuration "H-1" of Table 1, where the tag 140 can be an antibody-ssDNA barcode-enzyme with a chemical tag release mechanism. In the example configuration "H-1", the surface capture mechanism can be a DNA barcode hybridized with a complementary ssDNA, where the detected event 160 can be a polymerase activity using a DNA polymerase.

[0082] In a particular embodiment, the diagram 190 can represent the configuration "H-2" of Table 1, where the tag 140 can be an antibody-ssDNA barcode-enzyme with a photosensitive tag release mechanism. In the example configuration "H-2", the surface capture mechanism can be hybridization of the DNA barcode with complementary ssDNA, where the detected event 160 can be polymerase activity using a DNA polymerase.

[0083] In a particular embodiment, the diagram 190 can represent the configuration "H-3" of Table 1, where the tag 140 can be an antibody-ssDNA barcode-enzyme with an electrical tag release mechanism. In the example configuration "H-3", the surface capture mechanism can be hybridization of the DNA barcode to the complementary ssDNA, where the detected event 160 can be polymerase activity using a DNA polymerase.

[0084] In particular embodiments, event 160 may be detected via working electrode surface 124 and surface chemistry 122 .

[0085] Figure 2 An example flow chart 200 of sample preparation and separation and / or concentration chambers is shown. In certain embodiments, Figure 2 Flowchart 200 may describe a chamber and fluid channel architecture for sample preparation and separation / concentration chambers. In certain embodiments, flowchart 200 may begin at step 202, where a biological sample is placed within the system and transferred to a sample preparation chamber 204. As an example and not limitation, the sample preparation chamber may be subjected to a plasma purification process. As another example and not limitation, the sample preparation chamber may be subjected to conductivity modification. As an example and not limitation, separation and / or concentration may be achieved using DEP.

[0086] In certain embodiments, flow chart 200 may continue with transferring the biological sample to DEP chamber 208 , wherein DEP chamber waste 206 may be output from sample preparation chamber 204 , and DEP reagent input 209 may be input to sample preparation chamber 204 .

[0087] In certain embodiments, biological samples may be transferred from sample preparation chamber 204 to separation / concentration chamber 210. In certain embodiments, a variety of fluid reagents may be supplied to and from one or more fluid chambers using bulk and / or consumable reagent containers. In certain embodiments, a fluid guiding manifold and a power that is hydraulic, pneumatic, or electrostatic in nature may be used to supply a variety of fluid reagents to and from one or more fluid chambers. In instances where electrostatic forces are used, methods of electrowetting fluids using electrodes may be used, wherein electrodes may be positioned at the top or bottom of fluid channels and chambers, and wherein electrodes may be coated with a thin film selected to achieve hydrophobicity or hydrophilicity to support electrowetting.

[0088] In certain embodiments, DEP chamber waste 206 may be output from separation / concentration chamber 210, and DEP reagent input 209 may be input to separation / concentration chamber 210. In certain embodiments, a biological sample may be transferred from separation / concentration chamber 210 to labeling chamber 214. As an example and not limitation, one or more processing (e.g., labeling biomarkers) may occur within labeling chamber 214. In certain embodiments, DEP reagent input 209 may consist of one or more bulk reagents, such as bulk reagent Z' 218, bulk reagent B' 220, and / or bulk reagent A' 222.

[0089] Figure 3An example diagram 300 of a cross-sectional view of the dielectrophoresis concept is shown. In a particular embodiment, separation of one or more EVs 102 can be achieved in a detection device by using DEP, where DEP is a net force acting on particles with asymmetric polarizability in the presence of a radio frequency (RF) field. As shown in Figure 300, a plasma 320 of a biological sample can enter a DEP chamber (e.g., a separation and / or concentration chamber 210). As an example and not limitation, the plasma 320 can be composed of EVs 102 and various other biological particles 312, 314, 316. In this example, the EVs 102 can be measured as spherical objects of approximately 30-10,000 nm. As an example and not limitation, for any given combination of voltage, frequency, electrode spacing, and / or electrode geometry, a sub-range of the group of particles can be net attracted to the one or more electrodes. This phenomenon can be referred to as positive DEP. Particles that experience positive DEP will hereinafter be referred to as gravitational bands. As used herein, "gravitational bands" can refer to sub-ranges of EV 102 sizes. For the avoidance of doubt, it is clear that the gravity band is an adjustable range of EV 102 dimensions.

[0090] In certain embodiments, the one or more EVs 102 may experience an attractive force that is a function of the RF frequency, voltage, plasma conductivity, EV particle size, and EV particle charge when an alternating current (AC) waveform is passed through the positive electrode 304 and the negative electrode 306. It is understood that for any given voltage combination, a sub-range of particle sizes may produce a net attractive force on the one or more electrodes.

[0091] In this example, the positive electrode 304 and / or the negative electrode 306 may be made of metal or any other suitable material. In certain embodiments, the positive electrode 304 and / or the negative electrode 306 may be separated by the dielectric material 302. In certain embodiments, the EV 102 in the gravitational band may be attracted to one or more positive electrodes 304 and / or negative electrodes 306 on the surface of the dielectric material 302.

[0092] Figure 4An example diagram 400 of a dielectrophoresis electrode array is shown. In a particular embodiment, the one or more positive electrodes 304 and / or the one or more negative electrodes 306 may be arranged in an interdigitated configuration. As an example and not limitation, the interdigitated configuration may create one or more regions with steep electric field gradients. In this example, the length of the electrode 404 may be measured in hundreds (100) microns to millimeters, wherein the width of the electrode 402 may be less than one (1) micron. Similarly, the spacing and thickness of the electrodes may be less than one (1) micron. As an example and not limitation, sputtering may be used to form the electrode material. As another example and not limitation, alternative electrode deposition techniques may be used, including but not limited to atomic layer deposition, chemical vapor deposition, physical vapor deposition, electroforming, or any other suitable method. In a particular embodiment, the metal material of the positive electrode 304 and / or the negative electrode 306 may be any transition metal or noble metal. In practice, noble metals are advantageous, such as silver, gold, platinum, tungsten, ruthenium, rhodium, palladium, osmium, iridium, and / or any other suitable material.

[0093] In a specific embodiment, a polymer film is coated on the surface of the one or more positive electrodes 304 and / or one or more negative electrodes 306 and the dielectric 302. The polymer film is a multi-component mixture that is spin-cast onto the surface of the semiconductor wafer / chip. This material does not interfere with the electrical properties of the one or more electrodes, has a porosity that is adjusted for the attraction band, and is adjusted to be non-adsorbent. After casting, the film can be cured using UV light or thermal energy.

[0094] Figure 5A An example isometric diagram 500 of a generalized fluidic delivery method for a detection device is shown. In certain embodiments, the detection device can be a consumable chip (e.g., a printed circuit board) into which one or more biological samples (e.g., fluids) can be injected via one or more syringes 510 into the detection device.

[0095] In certain embodiments, one or more syringes 510 of the detection device can be actuated by one or more motors 520, wherein the motor 520 can contain one or more gears. In certain embodiments, the detection device can move a fluid (e.g., a biological sample) through a manifold 530 to a plurality of connected solenoid-actuated valves 540. It is understood that the solenoid-actuated valves can be arranged in parallel, in series, or in any other suitable configuration.

[0096] In a particular embodiment, the detection device can move a fluid through one or more reservoirs 550, wherein the one or more reservoirs 550 can be made of a centrifuge tube or any other suitable material. In a particular embodiment, the one or more syringes 510 of the detection device can be actuated by one or more motors 520, wherein the motor 520 can contain one or more gears. In a particular embodiment, the detection device can move a fluid (e.g., a biological sample) to a plurality of connected solenoid-actuated valves 540 through a manifold 530. It is understood that the solenoid-actuated valves can be arranged in parallel, in series, or in any other suitable configuration. In a particular embodiment, the detection device can move a fluid through one or more reservoirs 550, wherein the one or more reservoirs 550 can be made of a centrifuge tube or any other suitable material.

[0097] In certain embodiments, the detection device can move the fluid through one or more separation and / or labeling chambers 560, where labeling of the fluid and / or separation of the fluid can occur. As an example and not limitation, the one or more separation and / or labeling chambers 560 can label specific chemical groups.

[0098] In certain embodiments, the detection device can move fluid through one or more sensor chambers 570. By way of example and not limitation, the detection device can receive one or more biological samples via one or more reservoirs 550, where the biological samples can be transferred to one or more labeling and / or separation chambers 560 and then to one or more sensor chambers 570. Although this disclosure discusses a particular order in which biological samples are processed within a detection device, this disclosure contemplates any suitable order in which biological samples are processed within a detection device.

[0099] In certain embodiments, a specific bulk reagent may be dedicated to one or more specific chambers 560. As an example and not limitation, a chamber 560 may include a detector chamber, wherein the detector chamber may receive one or more "detector" reagents. As another example and not limitation, a specific detector reagent may be input to one or more specific chambers 560, or a specific detector reagent may be shared by all chambers 560. As an example and not limitation, each chamber in the one or more chambers 560 may indicate a specific biomarker. For example, one chamber 560 may indicate a specific biomarker, such as "biomarker 1", where another chamber 560 may be labeled "biomarker 2". In this example, one or more chambers 560 may cleave specific tags, where the tags may be chemically bound to one or more sensors and ultimately digitally quantified. In certain embodiments, the one or more chambers 560 may receive fluid (e.g., biological sample) input and output waste from each specific chamber.

[0100] Figure 5BAn example isometric diagram 580 of a generalized fluidic delivery method is shown. In certain embodiments, Figure 5A 500 of the detection device discussed above may be enclosed within a cartridge 590. As an example and not limitation, the cartridge 590 may house the one or more syringes 510, the one or more motors 520, the manifold 530, the one or more solenoid-actuated valves 540, the one or more reservoirs 550, the one or more chambers 560, and the one or more sensors 570.

[0101] Figure 6 An example diagram 600 of a device architecture for interrogating multiple EV-derived biomarkers 104 from one or more biological samples using fluidic multiplexing is shown. In particular embodiments, separation of biological samples can be performed in a single chamber, where a sample subvolume of the biological sample can be passed along the way to a biomarker-specific label and detection chamber (e.g., chamber 560). As an example and not limitation, one or more reservoirs 550 containing biological samples can be input into the device via a digital microfluidic sample input 610, where the one or more biological samples can be conveyed, flowed, or transferred to a dielectrophoresis (DEP) chamber 620. It is understood that the DEP chamber 620 can be the only chamber within the device of FIG. 600, or the DEP chamber 620 can be one of many DEP chambers 620 within the device of FIG. 600.

[0102] In certain embodiments, the DEP chamber 620 can process the one or more biological samples, wherein the biological samples can be transferred to the digital microfluidic (DMF) channel 630 until the biological samples reach one or more of the labeling chambers. It is understood that the DMF channel 630 can also be a microfluidic channel opposite to the digital microfluidic (DMF) channel. Figure 6As shown in FIG. 600 , the one or more biological samples may be processed and labeled in multiple chambers, such as a labeling chamber “1” 640, a labeling chamber “2” 642, and / or a labeling chamber “N” 644. In certain embodiments, the biological sample may be transferred from the one or more labeling chambers 640, 642, 644 to one or more sensor chambers 650, 652, and / or 654 via a DMF channel 630. In certain embodiments, a single, common detector chemistry may be used for each sensor chamber 650, 652, and / or 654. In this example, the chemistry may be common so that the specificity of the detected substance can be determined by the label, wherein after the one or more biological samples pass through the labeling chambers 640, 642, and / or 644, only the labeled exosomes of the biological sample may be detected. In this example, each of the N labeling chambers may only label one unique and specific biomarker. In particular embodiments, in one or more labeling chambers (e.g., labeling chambers 640, 642, 644), EV and / or biomarker-specific tags can be cleaved from EV 102 and subsequently chemically bound to one or more sensors (e.g., sensor 570), where the tags can be quantified digitally or by a suitable method.

[0103] Figure 7 An example diagram 700 of a device architecture for interrogating multiple EV-derived biomarkers 104 from one or more biological samples using fluidic multiplexing is shown. In certain embodiments, separation of one or more biological samples can be performed in a single chamber, where subvolumes of the one or more biological samples can be passed along the way to biomarker-specific labels and detection chambers. As an example and not limitation, one or more biological samples can be input into the device via a DMF sample input 610, where the one or more biological samples can be passed along the way to a DEP chamber 620. It is understood that the DEP chamber 620 can be the only chamber within the device of FIG. 700, or the DEP chamber 620 can be one of many DEP chambers 620 within the device of FIG. 700. It is understood that although the present disclosure discusses passing the one or more biological samples along the way to one or more chambers (e.g., DEP chamber 620), the present disclosure contemplates any suitable method for passing, transferring, and / or transporting the one or more biological samples along the way to one or more chambers and / or channels.

[0104] In certain embodiments, the DEP chamber 620 can process the one or more biological samples, wherein the biological samples can be transferred to the digital microfluidics (DMF) channel 630 until the biological samples reach one or more labeling chambers. Figure 7As shown in FIG. 700 , the one or more biological samples may be processed and assigned one or more labels in multiple chambers, such as label chamber “1” 640 and / or label chamber “N” 644. In a specific embodiment, the one or more biological samples may be transferred from one or more label chambers 640, 644 to one or more detector arrays 710, 720, 730 via a DMF channel 630. In a specific embodiment, each of the multiple detector arrays may be programmed to detect a specific biomarker. As an example and not limitation, the detector array 710 may be specifically programmed for a specific biomarker (such as “biomarker 1”). Similarly, the detector array 720 may be programmed for “biomarker 2”, while the detector array 730 may be programmed for “biomarker N”. In this example, each detector array may be programmed to run with a different detector chemistry, wherein each detector chamber may detect the attachment of a unique and specific designated biomarker. In particular embodiments, EV and / or biomarker specific tags may be cleaved from EV 102 in labeling chamber "1" 640, where the tags may be chemically bound to one or more sensors (e.g., detector arrays 710, 720, 730) and digitally quantified.

[0105] Figure 8 An example diagram 800 of a device architecture for interrogating multiple EV-derived biomarkers 104 from one or more biological samples using fluidic multiplexing is shown. In certain embodiments, separation of one or more biological samples can be performed in a single chamber, where sub-volumes of the one or more biological samples can be passed along the way to biomarker-specific labels and detection chambers. As an example and not limitation, one or more biological samples can be input into the device via a DMF sample input 610, where the one or more biological samples can be passed along the way to a DEP chamber 620. It is understood that the DEP chamber 620 can be the only chamber within the device of FIG. 800, or the DEP chamber 620 can be one of many DEP chambers 620 within the device of FIG. 800.

[0106] In certain embodiments, the DEP chamber 620 can process the one or more biological samples, wherein the one or more biological samples can be transferred to a digital microfluidic (DMF) channel 630 until the one or more biological samples reach one or more labeling chambers. As an example and not limitation, the DMF channel 630 can be a microfluidic channel. Figure 8As shown in FIG. 800 , the one or more biological samples may be processed and labeled by a single labeling chamber (such as labeling chamber “1” 640) in which the biological samples may be labeled and indicated. As an example and not limitation, the indicated sample 810 may be transferred from labeling chamber “1” 640 to one or more detector arrays 710, 720, 730 via a DMF channel 630 or a microfluidic channel. Figure 7 As discussed in , each of the multiple detector arrays can be programmed to detect a specific biomarker. As an example and not limitation, detector array 710 can be specifically programmed for a specific biomarker, such as "Biomarker 1". Similarly, detector array 720 can be programmed for "Biomarker 2", while detector array 730 can be programmed for "Biomarker N". Figure 8 In the example of FIG800, the volume of the designated EV (eg, designated sample 810) can pass through all detection chambers (eg, detector arrays 710, 720, 730), thereby improving the efficiency of using the biological sample. Figure 6-7 As discussed in , EV and / or biomarker specific tags can be lysed from EV 102 in labeling chamber "1" 640 and subsequently chemically bound to one or more sensors, where the tags can be digitally quantified.

[0107] Fig. 9 An example schematic diagram 900 showing the open circuit potential of a working electrode is shown. In certain embodiments, the circuit architecture of the device shown in FIG. 900 can be used to measure changes in the floating voltage of a working electrode 940 relative to a reference electrode 920 held at a potential by a counter electrode 930. By way of example and not limitation, the circuit of FIG. 900 can include a voltage source 950, a fluid cell 970, a digital-to-analog converter (DAC) 952, analog-to-digital converters 960, 962, diodes 954, 956, 958, a reference electrode 920, a counter electrode 930, and a working electrode 940. In certain embodiments, a method of preparing an electronic circuit for detecting one or more events as previously discussed can be comprised of a separate reference electrode (e.g., reference electrode 920), a counter electrode 930, and a working electrode 940 to detect immunochemical potential changes, immunochemical impedance changes, and / or immunochemical current changes (e.g., using an open circuit potential configuration, an electrochemical impedance spectroscopy configuration, a cyclic voltammetry configuration, or an ammeter configuration. Fig. 9 Figure 900 and Fig.10 1000 ). As an example and not limitation, the reference electrode 920, the counter electrode 930, and / or the working electrode 940 may be made of any metal, conductive metal oxide, or suitable conductive material (e.g., graphene), including but not limited to Pt, Au, Ag, Ag / AgCl, Zn, Ti, W, Pd, Ru, Pb, Cu, In, InxOy, ITO, AZO, ICO, graphene, etc.

[0108] In certain embodiments, each of the reference electrode 920, the counter electrode 930, and / or the working electrode 940 can be made of different materials. In certain embodiments, any one of the reference electrode 920, the counter electrode 930, and / or the working electrode 940 can be functionalized, while the remaining electrodes are not functionalized. As an example and not limitation, the working electrode 940 can be functionalized, while the reference electrode 920 and the counter electrode 930 can not be functionalized. In certain embodiments, the potential, impedance, and / or current at each of the reference electrode 920, the counter electrode 930, and / or the working electrode 940 can be monitored with one or more ADCs (e.g., ADC 960, 962), where the current can be stored to determine biomarker quantification.

[0109] Fig.10 An example schematic diagram 1000 showing an exemplary electrode configuration is shown. In certain embodiments, the circuit architecture of the device shown in FIG. 1000 can be used to measure the change in the floating voltage of the working electrode 940 relative to the reference electrode 920 held at a potential by the counter electrode 930. As an example and not limitation, the circuit of FIG. 1000 may include a voltage source 950, a digital-to-analog converter (DAC) 952, analog-to-digital converters 960, 962, 1020, operational amplifiers 1032, 1034, 1036, 1038, a reference electrode 920, a counter electrode 930, and a working electrode 940. In certain embodiments, a switch 1010 may be added to the open circuit potential, which may allow amperometric measurements and / or electrochemical impedance spectroscopy (EIS) to be performed.

[0110] Fig.11 An example diagram 1100 of an electrode configuration in which an embodiment may operate is shown. In a particular embodiment, a bottom printed circuit board (PCB) 1110 may be composed of a gasket 1132 and a working electrode 940. As an example and not limitation, a length 1150 of the bottom PCB 1110 may measure approximately 12.7 millimeters, and a width 1140 of the bottom PCB 1110 may measure approximately 12.7 millimeters. In a particular embodiment, a top PCB 1120 may be composed of a gasket 1132, one or more vents 1130, a reference electrode 920, and a counter electrode 1130. As an example and not limitation, a length 1150 of the top PCB 1120 may measure approximately 12.7 millimeters, and a width 1140 of the top PCB 1120 may measure approximately 12.7 millimeters. Although the present disclosure discusses a particular geometry, the present disclosure contemplates any suitable geometry, including elliptical, rectangular, and / or non-planar electrode configurations.

[0111] In a specific embodiment, a detection device (e.g., a device) for quantifying EV-derived biomarkers in a biological sample may include a device platform capable of accommodating a barrel. In a specific embodiment, a detection device (e.g., a device) for quantifying EV-derived biomarkers in one or more biological samples (e.g., blood, plasma, serum, cerebrospinal fluid, lymph, saliva, urine, feces, cell lysates, cell culture fluid) may include a device platform capable of accommodating a barrel and delivering samples, reagents, light and / or electrical pulses to multiple fluids and electrical channels on the barrel. As an example and not limitation, the device platform may include an electronic board with logic and / or power circuits for driving and sensing electronic devices, sensors and / or electrodes. As another example and not limitation, the device platform may include a fluid control system consisting of a fluid reservoir, a pipeline, and a pneumatic, hydraulic or electric fluid controller, wherein the fluid control controller may be manual and / or electronically actuated.

[0112] In a specific embodiment, a fluidics cartridge can be made of a molded top material and a molded bottom material, wherein the space between the top and bottom molded materials can accommodate one or more printed circuit boards and / or one or more microelectronic chips. As an example and not a limitation, the top molded material and the bottom molded material can be made of plastic or any other suitable material. In a specific embodiment, one or more adhesive layers can form a fluidic seal between the chamber and / or channel of the fluidics cartridge. In a specific embodiment, a "spacer" layer can define the boundary of a fluid chamber and / or channel together with the one or more printed circuit boards. As an example and not a limitation, the spacer can be made of plastic or any other suitable material. In a specific embodiment, the fluidics cartridge can include ports dedicated to the fluid inlet and outlet of each channel. In a specific embodiment, a component including the wall of one or more fluid chambers and / or channels can be coated with a non-biological fouling film. In a specific embodiment, one or more valves can be included to guide a variety of samples and / or reagents to various inlet and outlet ports and / or reservoirs and to guide a variety of samples and / or reagents from various inlet and outlet ports and / or reservoirs.

[0113] In certain embodiments, a detection device for quantifying EV-derived biomarkers in a biological sample may include a fluidic cartridge having multiple chambers and / or channels. In certain embodiments, a detection device for quantifying EV-derived biomarkers in a biological sample may include a printed circuit and / or one or more microelectronic silicon-based integrated chips (e.g., complementary metal oxide semiconductor (CMOS) or silicon-based integrated chips), wherein one or more chips may be configured to produce a DEP cavity (e.g., DEP chamber 620). As an example and not limitation, a DEP cavity may separate and capture particles of adjustable size and / or charge range from one or more biological samples (e.g., plasma samples).

[0114] In a particular embodiment, an apparatus for quantifying EV-derived biomarkers 104 in a plasma sample may include a printed circuit board and / or a silicon-based integrated chip with electrodes, wherein the printed circuit board and / or the silicon-based integrated chip can electrostatically control one or more fluidic motion systems. In a particular embodiment, an apparatus for quantifying EV-derived biomarkers in a plasma sample may include a fluid supply (e.g., sample input 610) and / or a waste management system. In a particular embodiment, an apparatus for quantifying EV-derived biomarkers in a biological sample may include one or more reaction chambers. As an example and not limitation, the one or more reaction chambers may include a DEP structure, wherein the DEP structure may support the marking and / or cleavage of EV-derived biomarker labels.

[0115] In certain embodiments, the apparatus for quantifying EV-derived biomarkers in a biological sample may include one or more printed circuits and / or microelectronic chips containing electrodes for electrically capturing and detecting labeled EVs and / or cleaved EV biomarker tags. In certain embodiments, the apparatus for quantifying EV-derived biomarkers 104 in a biological sample may include a plurality of electronic devices and / or sensors, wherein the electronic devices and / or sensors may report a digitized sensor response for the presence of the quantified biomarker 104 and / or biomarker tag.

[0116] In certain embodiments, the EV biosensor device can perform DEP particle filtration to separate EV 102 from other biological components, wherein the EV-biosensor device can be composed of one or more silicon chips. As an example and not limitation, the top of the one or more silicon chips can be composed of a metal layer, wherein the metal layer can contain an interdigitated alternating polarity electrode arrangement. In certain embodiments, the EV-biosensor device can contain an off-chip RF AC waveform generator, wherein the RF AC waveform generator can apply one or more specific waveforms on one or more electrodes (e.g., reference electrode 920, counter electrode 930, working electrode 940). In certain embodiments, the EV-biosensor device can include an anti-biofouling surface coating or other coating on the electrode and / or non-electrode surface within the DEP chamber 620.

[0117] Fig.12 An example schematic diagram 1200 showing an exemplary electrode configuration is shown. In a particular embodiment, the circuit architecture of the detection device shown in FIG. 1200 with the switch in the "on" position can be used to measure the change in the open circuit potential of the working electrode 940 relative to the reference electrode 920 held at a potential by the counter electrode 930. Alternatively, the circuit architecture of the device shown in FIG. 1200 with the switch in the "off" position can be used to measure the current flowing through the working electrode 940 in response to a defined voltage on the reference electrode 920. As an example and not limitation, the circuit of FIG. 1300 can include a digital-to-analog converter (DAC) 952, an analog-to-digital converter (ADC) 960, 962, 1020, operational amplifiers 1210, 1220, buffers 1230, 1240, a switch 1010, a reference electrode 920, a counter electrode 930, and a working electrode 940. In certain embodiments, the method of preparing an electronic circuit to detect one or more events as discussed above can be composed of an independent reference electrode (e.g., reference electrode 920), a counter electrode 930, and a working electrode 940 to detect immunochemical potential changes, immunochemical impedance changes, and / or immunochemical current changes using an open circuit potential configuration, an electrochemical impedance spectroscopy configuration, a cyclic voltammetry configuration, or an ammeter configuration. As an example and not limitation, the reference electrode 920, the counter electrode 930, and / or the working electrode 90 can be made of any metal, conductive metal oxide, or suitable conductive material.

[0118] In certain embodiments, each of the reference electrode 920, the counter electrode 940, and / or the working electrode 940 can be made of different materials. In certain embodiments, any one of the reference electrode 920, the counter electrode 930, and / or the working electrode 940 can be functionalized, while the remaining electrodes are not functionalized or are functionalized differently. In certain embodiments, the potential, impedance, and / or current at each of the reference electrode 920, the counter electrode 930, and / or the working electrode 940 can be monitored with one or more ADCs (e.g., ADC 960, 962, 1020), where the current can be stored to determine biomarker quantification.

[0119] Fig.13 An example diagram 1300 of an electrode configuration in which an embodiment may operate is shown. In a particular embodiment, the PCB 1330 may contain a bottom surface 1380, which may be composed of one or more working electrodes 940. As an example and not limitation, the length 1312 of the bottom surface 1380 may measure approximately twenty (20) millimeters. As another example and not limitation, the width 1310 of the bottom surface 1380 may measure approximately ten (10) millimeters. In a particular embodiment, the top surface 1382 and the bottom surface 1380 may be separated by one or more transfer adhesive layers 1392, which have one or more fluid ports 1340, 1350 and electrical pathways 1320.

[0120] In a particular embodiment, the top surface 1382 may be comprised of an electrical pathway 1320, one or more reference electrodes 920, and one or more counter electrodes 930. By way of example and not limitation, the length 1312 of the top surface 1382 may measure approximately twenty (20) millimeters, and the width 1310 of the top surface 1382 may measure approximately ten (10) millimeters. Although the present disclosure discusses the approximate length 1312 and width 1310 of the top surface 1382 and the bottom surface 1380, the present disclosure contemplates any suitable length 1312 and width 1310. Although the present disclosure discusses particular electrode configurations (e.g., the positioning of the counter electrode 930, the reference electrode 920, the working electrode 940), the present disclosure contemplates any suitable configuration of the counter electrode 930, the reference electrode 920, and / or the working electrode 940.

[0121] Fig.14An example method 1400 for separating and detecting biomarkers of interest via a dielectrophoresis device is shown. The method may begin at step 410, where a DEP device may receive a first biological sample containing one or more biomarkers of interest 104 onto a DEP electrode array of the DEP device. At step 1420, one or more processors of the DEP device may instruct the DEP device to apply a DEP force through one or more electrodes of the DEP electrode array. As an example and not limitation, one or more processors of the DEP device may instruct the DEP device to apply a DEP force of a specific intensity, direction, time period, or other configurable setting specifically for one or more biomarkers of interest 104 (e.g., EV 102) of the biological sample. In a particular embodiment, the DEP force may be an AC waveform. As an example and not limitation, the DEP force may be adjusted according to the size of the one or more target biomarkers 104.

[0122] In certain embodiments, the DEP electrode array may be arranged in an interdigitated configuration. In certain embodiments, the DEP electrode array may include a working electrode 940, a reference electrode 920, and / or a counter electrode 930. As an example and not limitation, the DEP electrode array may include one or more positive electrodes and one or more negative electrodes.

[0123] In certain embodiments, at step 1430, one or more sensors of the DEP device may determine the amount of one or more target biomarkers of interest 104 of the biological sample. As an example and not limitation, the sensor may quantify the total number of tags, labels, and / or markers corresponding to a particular biomarker 104. In certain embodiments, the amount of one or more particular biomarkers 104 may be output and presented via a GUI of one or more computing devices.

[0124] Where appropriate, specific implementations may be repeated Fig.14 Although the present disclosure describes and illustrates one or more steps of the method. Fig.14 The specific steps of the method occur in a specific order, but the present disclosure contemplates the steps occurring in any suitable order. Fig.14 Although the present disclosure describes and illustrates exemplary methods for separating and detecting one or more biomarkers of interest on a DEP device, including Fig.14 The present disclosure contemplates any suitable method for separating and detecting one or more biomarkers of interest on a DEP device, including any suitable steps, which may include, where appropriate Fig.14 In addition, although the present disclosure describes and illustrates the implementation of Fig.14specific components, devices or systems for specific steps of the method, but the present disclosure contemplates performing Fig.14 Any suitable combination of any suitable components, devices or systems of any suitable steps of the method.

[0125] Fig.15 An example method 1500 for detecting biomarkers of interest on a dielectrophoresis device is shown. The method may begin at step 1510, where a DEP device may receive a first biological sample containing one or more biomarkers of interest 104 onto one or more arrays of the DEP device. At step 1520, one or more processors of the DEP device may be instructed to apply a current through one or more electrode arrays. As an example and not limitation, one or more processors of the DEP device may instruct the DEP device to apply a DEP force of a specific intensity, direction, time period, or other configurable setting specifically for one or more biomarkers of interest 104 (e.g., EV 102) of the biological sample. In a particular embodiment, the DEP force may be an AC waveform. As an example and not limitation, the DEP force may be adjusted based on the size of the one or more target biomarkers 104.

[0126] In certain embodiments, the DEP electrode array may be arranged in an interdigitated configuration. In certain embodiments, the DEP electrode array may include a working electrode 940, a reference electrode 920, and / or a counter electrode 930. As an example and not limitation, the DEP electrode array may include one or more positive electrodes and one or more negative electrodes.

[0127] In certain embodiments, one or more biomarkers 104 may be isolated from a volume of a biological sample in response to a specific configuration of electrical current applied to one or more electrode arrays.

[0128] At step 1530, one or more processors of the DEP device may instruct the DEP device to filter one or more biomarkers 104 from a plurality of biomarkers 104 of the biological sample via one or more microfluidics. As an example and not limitation, the biomarkers 104 may be filtered into one or more chambers of the DEP device.

[0129] In certain embodiments, at step 1540, one or more processors of the DEP device may instruct the DEP device to label one or more biomarkers 104 of the biological sample with a marker. As an example and not limitation, a first biomarker may be labeled with a first marker, and a second biomarker may be labeled with a second marker. As another example and not limitation, the marker may be a plurality of tags, labels, or other forms of identifiers. In certain embodiments, the first marker may include a particular tag from a plurality of tags. As an example and not limitation, the plurality of tags may include an antibody tag, a metal nanoparticle (MNP) tag, a single-stranded DNA (ssDNA) tag, or any other suitable tag.

[0130] In certain embodiments, at step 1550, one or more sensors of one or more chambers of the DEP device may detect the amount of a specific biomarker 104. At step 1560, the DEP device may output and present a comprehensive result of the amount of the specific biomarker via the GUI of the computing device. In certain embodiments, one or more additional biological samples may be run on the DEP device simultaneously or in parallel.

[0131] Where appropriate, specific implementations may be repeated Fig.15 Although the present disclosure describes and illustrates one or more steps of the method. Fig.15 The specific steps of the method occur in a specific order, but the present disclosure contemplates the steps occurring in any suitable order. Fig.15 Although the present disclosure describes and illustrates exemplary methods for separating and detecting one or more biomarkers of interest on a DEP device, including Fig.15 The present disclosure contemplates any suitable method for separating and detecting one or more biomarkers of interest on a DEP device, including any suitable steps, which may include, where appropriate, Fig.15 In addition, although the present disclosure describes and illustrates the implementation of Fig.15 specific components, devices or systems for specific steps of the method, but the present disclosure contemplates performing Fig.15 Any suitable combination of any suitable components, devices or systems of any suitable steps of the method.

[0132] Fig.16An example method 1600 for separating at least one biomarker of interest on a device is shown. The method can begin at step 1610, where the device can receive a first biological sample. As an example and not limitation, the biological sample can contain one or more biomarkers 104. At step 1620, one or more processors of the device can instruct the device to transfer the biological sample to a first chamber via a microfluidic channel. As an example and not limitation, the first chamber can perform a specific treatment (e.g., a first treatment) on the first biological sample. In certain embodiments, the first chamber can include a sample preparation chamber, wherein the first treatment can change one or more characteristics of the biological sample. In certain embodiments, the first treatment can change the plasma purification of the first biological sample.

[0133] In certain embodiments, at step 1630, one or more processors of the DEP device may instruct the DEP device to transfer the first biological sample from the first chamber to the second chamber via one or more microfluidic channels. As an example and not limitation, a specific treatment may be performed on the first biological sample in the second chamber. The specific treatment performed on the first biological sample in the second chamber may be the same or different treatment as the treatment performed on the first biological sample in the first chamber. As an example and not limitation, the second chamber may include a concentrating chamber, wherein the current applied to the first biological sample in the concentrating chamber causes the ion concentration to change.

[0134] In certain embodiments, one or more processors of the DEP device may instruct the DEP device to transfer the first biological sample from the second chamber to the third chamber via the microfluidic channel at step 1640. As an example and not limitation, a third process may be performed on the first biological sample in the third chamber. In certain embodiments, the third chamber may include a separation chamber, wherein an electric current applied to the first biological sample in the separation chamber may result in separation of a specific biomarker of interest of the first biological sample.

[0135] In certain embodiments, one or more reagents may be input into any of the first, second, or third chambers. In certain embodiments, any of the first, second, or third chambers may output a waste product comprising a portion of the first biological sample and a portion of the one or more reagents produced by the first treatment. It is understood that the DEP device may repeat the treatment for multiple biological samples (e.g., a second biological sample, a third biological sample, etc.). In certain embodiments, one or more biological samples may be treated simultaneously or in parallel.

[0136] Where appropriate, specific implementations may be repeated Fig.16 Although the present disclosure describes and illustrates one or more steps of the method. Fig.16 The specific steps of the method occur in a specific order, but the present disclosure contemplates the steps occurring in any suitable order. Fig.16Although the present disclosure describes and illustrates example methods for separating and detecting one or more biomarkers of interest on a DEP device, including Fig.16 The present disclosure contemplates any suitable method for separating and detecting one or more biomarkers of interest on a DEP device, including any suitable steps, which may include, where appropriate, Fig.16 In addition, although the present disclosure describes and illustrates the implementation of Fig.16 specific components, devices or systems for specific steps of the method, but the present disclosure contemplates performing Fig.16 Any suitable combination of any suitable components, devices or systems of any suitable steps of the method.

[0137] Fig.17 An example method 1700 for separating at least one biomarker of interest on a solid-state device is shown. At step 1710, one or more processors of the solid-state device may provide instructions to receive a first biological sample into the solid-state device. As an example and not limitation, the first biological sample may be received via one or more microfluidic channels.

[0138] At step 1720, one or more processors of the solid-state device may instruct one or more components of the solid-state device to transfer the first biological sample to the DEP chamber. As an example and not limitation, one or more processes may be performed on the first biological sample in the DEP chamber. In a particular embodiment, the DEP chamber may contain one or more electrode arrays. As an example and not limitation, each of the one or more electrode arrays may be arranged in an interdigitated configuration. As another example and not limitation, each of the one or more electrode arrays may include a working electrode 940, a reference electrode 920, and a counter electrode 930.

[0139] In certain embodiments, at step 1730, one or more processors of the solid-state device may instruct one or more components of the solid-state device to transfer the first biological sample to the labeling chamber through a microfluidic (MF) channel.

[0140] In certain embodiments, at step 1740, one or more processors of the solid-state device may instruct one or more components of the solid-state device to process the first biological sample in the labeling chamber. As an example and not limitation, one or more biomarkers of interest of the first biological sample may be labeled based on one or more features. In certain embodiments, one or more biomarkers of interest of the first biological sample may be labeled with a specific label from a plurality of labels. As an example and not limitation, the plurality of labels may include at least an antibody label, a metal nanoparticle (MNP) label, and / or a single-stranded DNA (ssDNA) label.

[0141] In particular embodiments, at step 1750, one or more processors of the solid-state device may instruct one or more components of the solid-state device to transfer one or more biomarkers of interest to the sensor chamber.

[0142] In particular embodiments, at step 1760, one or more processors of the solid-state device may instruct one or more components of the solid-state device to digitally determine the amount of the one or more biomarkers of interest within the sensor chamber based on the amount of label within the first biological sample.

[0143] It is understood that the solid-state device can repeat the process for multiple biological samples (eg, a second biological sample, a third biological sample, etc.) In certain embodiments, one or more biological samples can be processed simultaneously or in parallel.

[0144] Specific embodiments may be repeated where appropriate. Fig.17 Although the present disclosure describes and illustrates one or more steps of the method. Fig.17 The specific steps of the method occur in a specific order, but the present disclosure contemplates the steps occurring in any suitable order. Fig.17 Although the present disclosure describes and illustrates example methods for separating and detecting one or more biomarkers of interest on a DEP device, including Fig.17 The present disclosure contemplates any suitable method for separating and detecting one or more biomarkers of interest on a solid-state device, including any suitable steps, which may include, where appropriate, Fig.17 In addition, although the present disclosure describes and illustrates the implementation of Fig.17 specific components, devices or systems for specific steps of the method, but the present disclosure contemplates performing Fig.17 Any suitable combination of any suitable components, devices or systems of any suitable steps of the method.

[0145] Fig.18 An example method 1800 for separating at least one biomarker of interest on a solid-state device is shown. In step 1810, one or more processors of the solid-state device may provide instructions to receive a first biological sample into the solid-state device. As an example and not limitation, the first biological sample may include one or more biomarkers of interest. As another example and not limitation, the first biological sample may be input via a microfluidic (MF) channel.

[0146] At step 1820, one or more processors of the solid-state device may provide instructions to transfer the first biological sample to the DEP chamber via the MF and apply an AC current to the first biological sample through one or more electrodes of the DEP chamber. In a particular embodiment, the one or more electrodes of the DEP chamber may include at least a working electrode 940, a reference electrode 920, and a counter electrode 930.

[0147] At step 1830, one or more processors of the solid-state device may provide instructions to one or more components of the solid-state device to transfer the first biological sample from the DEP chamber to the labeling chamber from the MF.

[0148] In step 1840, one or more processors of the solid-state device may provide instructions to one or more components of the solid-state device to assign a label to each of the one or more biomarkers of interest of the first biological sample. As an example and not limitation, each label may define a specific label in a plurality of labels. As another example and not limitation, a plurality of labels may include, but are not limited to, antibody labels, metal nanoparticle (MNP) labels, or single-stranded DNA (ssDNA) labels. In a particular embodiment, a first label may be assigned to a first biomarker of interest of the first biological sample based on one or more specific features. In a particular embodiment, a second label may be assigned to a second biomarker of interest of the first biological sample based on one or more specific features.

[0149] At step 1850, one or more processors of the solid-state device may provide instructions to one or more components of the solid-state device to transfer the first biological sample to one or more detector chambers via MF, wherein each detector chamber is programmed for a specific biomarker of interest.

[0150] In step 1860, one or more processors of the solid-state device may provide instructions to one or more components of the solid-state device to digitally determine the amount of the one or more biomarkers of interest based on the assigned labels via one or more detectors within one or more detector chambers.

[0151] It is understood that the solid-state device can repeat the process for multiple biological samples (eg, a second biological sample, a third biological sample, etc.) In certain embodiments, one or more biological samples can be processed simultaneously or in parallel.

[0152] Specific embodiments may be repeated where appropriate. Fig.18 Although the present disclosure describes and illustrates one or more steps of the method. Fig.18 The specific steps of the method occur in a specific order, but the present disclosure contemplates the steps occurring in any suitable order. Fig.18Although the present disclosure describes and illustrates example methods for separating and detecting one or more biomarkers of interest on a solid-state device, including Fig.18 The present disclosure contemplates any suitable method for separating and detecting one or more biomarkers of interest on a solid-state device, including any suitable steps, which may include, where appropriate, Fig.18 In addition, although the present disclosure describes and illustrates the implementation of Fig.18 specific components, devices or systems for specific steps of the method, but the present disclosure contemplates performing Fig.18 Any suitable combination of any suitable components, devices or systems of any suitable steps of the method.

[0153] System and method

[0154] Fig.19 An example computer system 1900 is shown, which can be used to perform digital, multiplexed, extracellular vesicle-derived biomarker chip lab diagnostics according to the currently disclosed embodiments. In a specific embodiment, one or more computer systems 1900 perform one or more steps of one or more methods described or shown herein. In a specific embodiment, one or more computer systems 1900 provide the functions described or shown herein. In a specific embodiment, software running on one or more computer systems 1900 performs one or more steps of one or more methods described or shown herein, or provides the functions described or shown herein. Specific embodiments include one or more parts of one or more computer systems 1900. Herein, references to computer systems may cover computing devices, and vice versa, where appropriate. In addition, references to computer systems may cover one or more computer systems, where appropriate.

[0155] The present disclosure contemplates any suitable number of computer systems 1900. The present disclosure contemplates computer systems 1900 in any suitable physical form. As examples and not limitations, computer system 1900 may be an embedded computer system, a system on a chip (SOC), a single board computer system (SBC) (e.g., a computer on a module (COM) or a system on a module (SOM)), a desktop computer system, a laptop or notebook computer system, an interactive kiosk, a mainframe computer, a computer system grid, a mobile phone, a personal digital assistant (PDA), a server, a tablet computer system, an augmented / virtual reality device, or a combination of two or more thereof. Where appropriate, computer system 1900 may include one or more computer systems 1900; it may be stand-alone or distributed; span multiple locations; span multiple machines; span multiple data centers; or reside in a cloud, which may include one or more cloud components in one or more networks.

[0156] Where appropriate, one or more computer systems 1900 may perform one or more steps of one or more methods described or illustrated herein without substantial spatial or temporal limitations. As an example and not limitation, one or more computer systems 1900 may perform one or more steps of one or more methods described or illustrated herein in real time or batch mode. Where appropriate, one or more computer systems 1900 may perform one or more steps of one or more methods described or illustrated herein at different times or at different locations.

[0157] In a particular embodiment, computer system 1900 includes processor 1902, memory 1904, storage 1906, input / output (I / O) interface 1908, communication interface 1910, and bus 1912. Although the present disclosure describes and illustrates a particular computer system having a particular number of particular components in a particular arrangement, the present disclosure contemplates any suitable computer system having any suitable number of any suitable components in any suitable arrangement. In a particular embodiment, processor 1902 includes hardware for executing instructions (such as those that constitute a computer program). As an example and not limitation, to execute instructions, processor 1902 may retrieve (or fetch) instructions from internal registers, internal caches, memory 1904, or storage 1906; decode and execute them; and then write one or more results to internal registers, internal caches, memory 1904, or storage 1906. In a particular embodiment, processor 1902 may include one or more internal caches for data, instructions, or addresses. The present disclosure contemplates that processor 1902 includes any suitable number of any suitable internal caches where appropriate. As an example and not limitation, the processor 1902 may include one or more instruction caches, one or more data caches, and one or more translation lookaside buffers (TLBs). The instructions in the instruction cache may be a copy of the instructions in the memory 1904 or storage 1906, and the instruction cache may speed up the retrieval of these instructions by the processor 1902.

[0158] The data in the data cache may be a copy of data in memory 1904 or storage 1906 for instructions for executing operations at processor 1902; access for subsequent instructions executed at processor 1902 or results of previous instructions executed at processor 1902 for writing to memory 1904 or storage 1906; or other suitable data. The data cache may speed up read or write operations of processor 1902. The TLB may speed up virtual address translation of processor 1902. In a particular embodiment, processor 1902 may include one or more internal registers for data, instructions, or addresses. The present disclosure contemplates that processor 1902 includes any suitable number of any suitable internal registers, where appropriate. Where appropriate, processor 1902 may include one or more arithmetic logic units (ALUs); be a multi-core processor; or include one or more processors 1902. Although the present disclosure describes and illustrates a particular processor, the present disclosure contemplates any suitable processor.

[0159] In a particular embodiment, memory 1904 includes main memory for storing instructions executed by processor 1902 or data operated by processor 1902. As an example and not limitation, computer system 1900 can load instructions from memory 1906 or another source (such as, for example, another computer system 1900) to memory 1904. Processor 1902 can then load instructions from memory 1904 to internal registers or internal caches. To execute the instructions, processor 1902 can retrieve the instructions from the internal registers or internal caches and decode them. During or after executing the instructions, processor 1902 can write one or more results (which can be intermediate results or final results) to the internal registers or internal caches. Processor 1902 can then write one or more of those results to memory 1904. In a particular embodiment, processor 1902 only executes instructions in one or more internal registers or internal caches or in memory 1904 (rather than memory 1906 or elsewhere), and only operates on data in one or more internal registers or internal caches or in memory 140 (rather than memory 1906 or elsewhere).

[0160] One or more memory buses (each of which may include an address bus and a data bus) may couple the processor 1902 to the memory 1904. The bus 1912 may include one or more memory buses, as described below. In a particular embodiment, one or more memory management units (MMUs) are located between the processor 1902 and the memory 1904 and facilitate the processor 1902 to request access to the memory 1904. In a particular embodiment, the memory 1904 includes a random access memory (RAM). Where appropriate, the RAM may be a volatile memory. Where appropriate, the RAM may be a dynamic RAM (DRAM) or a static RAM (SRAM). In addition, where appropriate, the RAM may be a single-port or multi-port RAM. The present disclosure contemplates any suitable RAM. Where appropriate, the memory 1904 may include one or more memory devices 104. Although the present disclosure describes and illustrates a specific memory, the present disclosure contemplates any suitable memory.

[0161] In a particular embodiment, the memory 1906 includes a large capacity memory for data or instructions. As an example and not limitation, the memory 1906 may include a hard disk drive (HDD), a floppy disk drive, a flash memory, an optical disk, a magneto-optical disk, a tape, or a universal serial bus (USB) drive, or a combination of two or more thereof. Where appropriate, the memory 1906 may include a removable or non-removable (or fixed) medium. Where appropriate, the memory 1906 may be internal or external to the computer system 1900. In a particular embodiment, the memory 1906 is a non-volatile solid-state memory. In a particular embodiment, the memory 1906 includes a read-only memory (ROM). Where appropriate, the ROM may be a mask-programmed ROM, a programmable ROM (PROM), an erasable PROM (EPROM), an electrically erasable PROM (EEPROM), an electrically alterable ROM (EAROM), or a flash memory, or a combination of two or more thereof. The present disclosure contemplates a large capacity memory 1906 in any suitable physical form. Where appropriate, the memory 1906 may include one or more memory control units, which facilitate communication between the processor 1902 and the memory 1906. Where appropriate, memory 1906 may include one or more memories 1906. Although this disclosure describes and illustrates particular memories, this disclosure contemplates any suitable memories.

[0162] In a particular embodiment, the I / O interface 1908 includes hardware, software, or both, providing one or more interfaces for communication between the computer system 1900 and one or more I / O devices. Where appropriate, the computer system 1900 may include one or more of these I / O devices. One or more of these I / O devices can enable communication between a person and the computer system 1900. As an example and not limitation, the I / O device may include a keyboard, a keypad, a microphone, a monitor, a mouse, a printer, a scanner, a speaker, a static camera, a stylus, a tablet computer, a touch screen, a trackball, a camera, other suitable I / O devices, or a combination of two or more thereof. The I / O device may include one or more sensors. The present disclosure contemplates any suitable I / O device and any I / O interface 1906 suitable for them. Where appropriate, the I / O interface 1908 may include one or more devices or software drivers so that the processor 1902 can drive one or more of these I / O devices. Where appropriate, the I / O interface 1908 may include one or more I / O interfaces 1906. Although this disclosure describes and illustrates a particular I / O interface, this disclosure contemplates any suitable I / O interface.

[0163] In particular embodiments, the communication interface 1910 includes hardware, software, or both that provides one or more interfaces for communicating (such as, for example, packet-based communications) between the computer system 1900 and one or more other computer systems 1900 or one or more networks. As an example and not limitation, the communication interface 1910 may include a network interface controller (NIC) or network adapter for communicating with an Ethernet or other wired network, or a wireless NIC (WNIC) or wireless adapter for communicating with a wireless network (such as a WI-FI network). The present disclosure contemplates any suitable network and any communication interface 1910 suitable therefor.

[0164] As an example and not limitation, the computer system 1900 may communicate with one or more parts of a self-organizing network, a personal area network (PAN), a local area network (LAN), a wide area network (WAN), a metropolitan area network (MAN), or the Internet, or a combination of two or more thereof. One or more parts of one or more of these networks may be wired or wireless. For example, the computer system 1900 may communicate with a wireless PAN (WPAN) (such as, for example, a Bluetooth WPAN), a WI-FI network, a WI-MAX network, a cellular phone network (such as, for example, a global system for mobile communications (GSM) network), or other suitable wireless networks, or a combination of two or more thereof. Where appropriate, the computer system 1900 may include any suitable communication interface 1910 for any of these networks. Where appropriate, the communication interface 1910 may include one or more communication interfaces 1910. Although the present disclosure describes and illustrates a specific communication interface, the present disclosure contemplates any suitable communication interface.

[0165] In a particular embodiment, bus 1912 includes hardware, software, or both that couples components of computer system 1900 to each other. As an example and not limitation, bus 1912 may include an accelerated graphics port (AGP) or other graphics bus, an enhanced industry standard architecture (EISA) bus, a front side bus (FSB), a HYPERTRANSPORT (HT) interconnect, an industry standard architecture (ISA) bus, an INFINIBAND interconnect, a low pin count (LPC) bus, a memory bus, a micro channel architecture (MCA) bus, a peripheral component interconnect (PCI) bus, a PCI-Express (PCIe) bus, a serial advanced technology attachment (SATA) bus, a video electronics standard association local (VLB) bus, or other suitable bus or a combination of two or more thereof. Where appropriate, bus 1912 may include one or more buses 1912. Although the present disclosure describes and illustrates a particular bus, the present disclosure contemplates any suitable bus or interconnect.

[0166] Miscellaneous

[0167] As used herein, "or" is inclusive and not exclusive, unless expressly stated otherwise or the context indicates otherwise. Thus, "A or B" herein means "A, B, or both," unless expressly stated otherwise or the context indicates otherwise. Additionally, "and" means both jointly and severally, unless expressly stated otherwise or the context indicates otherwise. Thus, "A and B" herein means "A and B, jointly or severally," unless expressly stated otherwise or the context indicates otherwise.

[0168] As used herein, "automatically" and its derivatives mean "without human intervention" unless expressly stated otherwise or the context indicates otherwise.

[0169] The embodiments disclosed herein are only examples, and the scope of the present disclosure is not limited to them. The embodiments according to the present invention are particularly disclosed in the attached claims relating to methods, storage media, systems and computer program products, wherein any feature mentioned in one claim category, such as a method, can also be claimed in another claim category, such as a system. The dependencies or references returned to the attached claims are selected only for formal reasons. However, any subject matter caused by intentionally returning to any previous claim (especially multiple dependencies) can also be claimed, so that any combination of claims and their features can be disclosed and claimed, regardless of the dependencies selected in the attached claims. The subject matter that can be claimed includes not only the combination of features as set forth in the attached claims, but also any other combination of features in the claims, wherein each feature mentioned in the claims can be combined with any other feature in the claims or a combination of other features. In addition, any of the embodiments and features described or depicted herein can be claimed in a separate claim and / or with any embodiment or feature described or depicted herein or any one of the features of the attached claims in any combination.

[0170] The scope of the present disclosure covers all changes, substitutions, variations, alterations and modifications to the example embodiments described or shown herein that will be understood by those skilled in the art. The scope of the present disclosure is not limited to the example embodiments described or shown herein. In addition, although the present disclosure describes and illustrates that the various embodiments herein include specific parts, elements, features, functions, operations or steps, any of these embodiments may include any combination or arrangement of any parts, elements, features, functions, operations or steps described or shown anywhere herein that will be understood by those skilled in the art. In addition, in the appended claims, a device or system or a component of a device or system that is suitable, arranged, capable, configured, enabled, operable or effectively performs a specific function is mentioned, covering the device, system, component, whether it or the specific function is activated, turned on or unlocked, as long as the device, system or component is so suitable, arranged, capable, configured, enabled, operable or effective. In addition, although the present disclosure describes or illustrates that a specific embodiment provides specific advantages, a specific embodiment may not provide some or all of these advantages.

Claims

1. A method for separating and detecting one or more biomarkers of interest on a dielectrophoresis device, comprising: receiving a first biological sample containing the one or more biomarkers of interest onto a dielectrophoresis (DEP) electrode array; applying a DEP force through specific electrodes of a DEP electrode array, wherein the strength, direction, and time duration of the DEP force are specific to the one or more biomarkers of interest of the first biological sample; as well as The amount of the one or more biomarkers of interest of the first biological sample is determined via a digital sensor.

2. The method according to claim 1, wherein: The DEP electrode array is arranged in an interdigitated configuration.

3. The method according to claim 1, wherein: The DEP electrode array includes at least a working electrode, a reference electrode and a counter electrode.

4. The method according to claim 1, wherein: The DEP electrode array includes one or more positive electrodes and one or more negative electrodes.

5. The method according to claim 1, wherein: The DEP force is an alternating current (AC) waveform.

6. The method according to claim 1, wherein: One biomarker of interest is extracellular vesicles (EVs).

7. The method according to claim 6, wherein: The DEP force is adjusted according to the size of the EV particles.

8. A dielectrophoresis device for separating and detecting one or more biomarkers of interest, the dielectrophoresis device comprising: means for receiving a first biological sample containing the one or more biomarkers of interest onto a dielectrophoresis (DEP) electrode array; means for applying a DEP force through specific electrodes of a DEP electrode array, wherein the intensity, direction, and time duration of the DEP force are specific to the one or more biomarkers of interest of the first biological sample; as well as Means for determining the amount of said one or more biomarkers of interest of said first biological sample via a digital sensor.

9. A method for detecting at least one biomarker of interest on a dielectrophoresis (DEP) device, the method comprising: Receiving a first biological sample onto one or more electrode arrays of a DEP device, wherein the first biological sample contains one or more biomarkers of interest; applying an electric current through the one or more electrode arrays to separate a first biomarker of interest from a plurality of biomarkers of interest in the first biological sample; filtering the first biomarker of interest from a plurality of biomarkers of the first biological sample into one or more chambers of the DEP device via one or more microfluidics; labeling the first biomarker of interest with a first marker; detecting the amount of the first biomarker of interest via an electrochemical sensor; and Displays the overall results.

10. The method according to claim 9, wherein: Each of the one or more electrode arrays includes at least a working electrode, a reference electrode, and a counter electrode.

11. The method according to claim 9, wherein: The first marker is a specific tag among a plurality of tags.

12. The method according to claim 11, wherein: The plurality of tags at least include antibody tags, metal nanoparticle (MNP) tags or single-stranded DNA (ssDNA) tags.

13. The method according to claim 9, wherein: The one or more electrode arrays are arranged in an interdigitated configuration.

14. The method according to claim 9, wherein: Each of the one or more electrode arrays includes one or more positive electrodes and one or more negative electrodes.

15. The method according to claim 9, wherein: The first biomarker of interest is extracellular vesicles (EVs).

16. A dielectrophoresis device for detecting at least one biomarker of interest, the dielectrophoresis device comprising: means for receiving a first biological sample onto one or more electrode arrays of the dielectrophoresis device, wherein the first biological sample contains one or more biomarkers of interest; means for applying an electric current through the one or more electrode arrays to separate a first biomarker of interest from a plurality of biomarkers of interest in the first biological sample; means for filtering the first biomarker of interest from a plurality of biomarkers of the first biological sample into one or more chambers of the dielectrophoresis device via one or more microfluidics; means for labeling said first biomarker of interest with a first marker; means for detecting the amount of said first biomarker of interest via an electrochemical sensor; and Tools for displaying synthesis results.

17. A method for detecting at least one biomarker of interest on a device, the method comprising: receiving a first biological sample; transferring the first biological sample to a first chamber via a microfluidic channel, wherein a first process is performed on the first biological sample in the first chamber; transferring the first biological sample from the first chamber to a second chamber via the microfluidic channel, wherein a second process is performed on the first biological sample in the second chamber; and The first biological sample is transferred from the second chamber to a third chamber via the microfluidic channel, wherein a third process is performed on the first biological sample in the third chamber.

18. The method according to claim 17, wherein: The first chamber comprises a sample preparation chamber, wherein the first treatment changes one or more properties of the first biological sample.

19. The method according to claim 18, wherein: The first treatment alters the plasma purification of the first biological sample.

20. The method of claim 17, further comprising: introducing one or more reagents into the first chamber; as well as A waste product is output from the first chamber, the waste product comprising a portion of the first biological sample and a portion of the one or more reagents produced by the first processing.

21. The method according to claim 17, wherein: The second chamber includes a concentrating chamber, wherein an electric current applied to the first biological sample in the concentrating chamber causes a change in ion concentration.

22. The method according to claim 17, wherein: The third chamber comprises a separation chamber, wherein an electric current applied to the first biological sample in the separation chamber results in separation of a specific biomarker of interest of the first biological sample.

23. A device for detecting at least one biomarker of interest, the device comprising: means for receiving a first biological sample; means for transferring the first biological sample to a first chamber via a microfluidic channel, wherein a first process is performed on the first biological sample in the first chamber; means for transferring the first biological sample from the first chamber to a second chamber via the microfluidic channel, wherein a second process is performed on the first biological sample in the second chamber; and Means for transferring the first biological sample from the second chamber to a third chamber via the microfluidic channel, wherein a third process is performed on the first biological sample in the third chamber.

24. A method for separating at least one biomarker of interest on a solid state device, the method comprising: receiving a first biological sample into the solid-state device, transferring the first biological sample to a dielectrophoresis (DEP) chamber, wherein one or more processes are performed on the first biological sample in the DEP chamber; transferring the first biological sample to a labeling chamber through a microfluidic (MF) channel; processing the first biological sample in the labeling chamber, wherein one or more biomarkers of interest of the first biological sample are labeled based on one or more characteristics; delivering the one or more biomarkers of interest with corresponding labels to the sensor chamber; and The amount of the one or more biomarkers of interest having the corresponding labels is digitally determined by one or more processes performed in the sensor chamber.

25. The method according to claim 24, wherein: The first biological sample is received via the microfluidic channel.

26. The method according to claim 24, wherein: The one or more biomarkers of interest of the first biological sample are labeled with a specific tag from a plurality of tags.

27. The method according to claim 26, wherein: The plurality of tags at least include antibody tags, metal nanoparticle (MNP) tags or single-stranded DNA (ssDNA) tags.

28. The method of claim 24, wherein: The DEP chamber includes one or more electrode arrays.

29. The method according to claim 28, wherein: Each of the one or more electrode arrays is arranged in an interdigitated configuration.

30. The method of claim 29, wherein: Each of the one or more electrode arrays includes at least a working electrode, a reference electrode, and a counter electrode.

31. A solid-state device for separating at least one biomarker of interest, the device comprising: means for receiving a first biological sample into said solid-state device, means for transferring the first biological sample to a dielectrophoresis (DEP) chamber, wherein one or more processes are performed on the first biological sample in the DEP chamber; means for transferring said first biological sample to a labeling chamber through a microfluidic (MF) channel; means for processing the first biological sample in the labeling chamber, wherein one or more biomarkers of interest of the first biological sample are labeled based on one or more characteristics; means for delivering the one or more biomarkers of interest with corresponding labels to the sensor chamber; and Means for digitally determining the amount of said one or more biomarkers of interest having corresponding labels by one or more processes performed in said sensor chamber.

32. A method for separating at least one biomarker of interest on a solid state device, the method comprising: receiving a first biological sample into the solid-state device, wherein the first biological sample comprises one or more biomarkers of interest and the first biological sample is input via a microfluidic (MF) channel; transferring the first biological sample to a dielectrophoresis (DEP) chamber via MF and applying an alternating current (AC) to the first biological sample through one or more electrodes of the DEP chamber; transferring the first biological sample from the DEP chamber to a labeling chamber via MF; assigning a label to each of the one or more biomarkers of interest for the first biological sample; transferring the first biological sample via the MF to one or more detector chambers, wherein each detector chamber is programmed for a specific biomarker of interest; and The amount of the one or more biomarkers of interest is digitally determined by one or more detectors within the one or more detector chambers based on the assigned labels.

33. The method of claim 32, wherein: The tag defines a specific tag among a plurality of tags.

34. The method of claim 33, wherein: The plurality of tags at least include antibody tags, metal nanoparticle (MNP) tags or single-stranded DNA (ssDNA) tags.

35. The method of claim 32, wherein: A first biomarker of interest in the first biological sample is assigned a first label based on a specific characteristic.

36. The method of claim 32, wherein: A second biomarker of interest in the first biological sample is assigned a second label based on the specific characteristic.

37. The method of claim 32, wherein: The one or more electrodes of the DEP chamber include at least a working electrode, a reference electrode, and a counter electrode.

38. A solid-state device for separating at least one biomarker of interest, the solid-state device comprising: means for receiving a first biological sample into the solid-state device, wherein the first biological sample comprises one or more biomarkers of interest and the first biological sample is input via a microfluidic (MF) channel; means for transferring the first biological sample to a dielectrophoresis (DEP) chamber via the MF channel and applying an alternating current (AC) to the first biological sample through one or more electrodes of the DEP chamber; means for transferring the first biological sample from the DEP chamber to a labeling chamber via the MF channel; means for assigning a label to each of said one or more biomarkers of interest of said first biological sample; means for transferring the first biological sample via the MF channel to one or more detector chambers, wherein each detector chamber is programmed for a specific biomarker of interest; and Means for digitally determining the amount of the one or more biomarkers of interest based on the assigned labels via one or more detectors within the one or more detector chambers.

39. A solid-state device comprising: syringe; a microfluidic channel, wherein the microfluidic channel receives a biological sample; one or more motors, each of which includes one or more gears; One or more actuating valves, wherein each of the one or more actuating valves is connected to a plurality of the microfluidic channels; one or more reservoirs, wherein the one or more reservoirs are connected to the microfluidic channel; one or more dielectrophoresis (DEP) chambers, wherein each DEP chamber contains at least one or more working electrodes, one or more reference electrodes, and one or more counter electrodes; one or more labeling chambers, wherein the biological sample is assigned one or more labels; one or more detector chambers wherein the biological sample and associated one or more markers are processed; and One or more detectors, wherein the one or more detectors are operable to quantify the one or more markers.

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