Method and device for detecting the concentration of at least one analyte in a sample

Abstract

This invention relates to a method for detecting the concentration of an analyte (10) in a sample (20), the method comprising: providing a device (100) having a channel (40), the channel (40) having a substrate (42), the substrate (42) having a surface binder element (44) and / or a surface test analyte (47); mixing a measuring element (30) into the sample (20), the measuring element (30) having a capture binder element (35) and / or the measuring element (30) having an element test analyte (38); allowing the sample (20) to flow through the channel (40); moving the measuring element (30) to the substrate (42); moving the measuring element (30) along the substrate (42) / on the substrate (42) and by means of interaction modification The invention relates to a device (100) for varying the velocity of at least one measuring element (30), the interaction comprising one or more of the following: interaction between at least one surface binder element (44), the analyte (10) and at least one trap binder element (35); interaction between a surface test analyte (47) and at least one of the trap binder elements (35); and / or interaction between an element test analyte (38) of the at least one measuring element (30) and at least one surface binder element (44); and determining the concentration of the analyte (10) by determining the velocity change of the measuring element (30) and / or by determining the loss of the measuring element (30), and / or by determining the velocity change of the measuring element (30).

Description

Technical Field

[0001] The present invention relates to a method for detecting the concentration of at least one analyte in a sample, and an apparatus for detecting the concentration of at least one analyte in a sample. Background of the Invention

[0003] In clinical chemistry, the detection and quantification of extracellular vesicles (EVs) and other vesicle analytes are challenging tasks. Performing these assays requires significant manual labor (lacking standardization), ultracentrifugation, and substantial capital investment in central laboratory equipment. However, these assays cannot be performed with unprocessed patient samples (e.g., whole blood). They typically utilize expensive optically labeled signal readout devices (e.g., for fluorescence flow cytometry) and cannot be integrated into point-of-use workflows due to their reliance on workflow-disrupting centrifugation. Furthermore, the typical dynamic concentration range of reported assays is on the order of 2–3 logarithmic scales, which is insufficient to simultaneously cover multiple vesicle biomarkers for multiplex analysis.

[0004] Such measurements can be classified as heterogeneous measurements, homogeneous measurements, or competitive measurements.

[0005] Heterogeneous assays are a class of biochemical analyses that involve separating unbound components from bound target analytes. In these assays, target molecules (e.g., proteins or nucleic acids) are immobilized on a solid support (e.g., microbeads or microplate wells), while the unbound components are washed away. Detection of the bound analyte is achieved using labeled probes (e.g., antibodies or fluorescent molecules) that specifically bind to the immobilized target. This binding event can be quantified using various detection methods, such as fluorescence, luminescence, or absorbance measurements. These workflows always require a sequence of operations including a washing step.

[0006] Homogeneous assays do not require a separation step to detect the target analyte. These assays are performed in a homogeneous solution without the need for washing or removal of unbound components. Instead, the detection signal is generated by the specific interaction between the target analyte and the two antibodies under close-range conditions, and is used for fluorescence resonance energy transfer (FRET) readout. However, sufficient optical transparency is required so that the FRET effect is not distorted.

[0007] Competitive assays are a class of biochemical analyses involving competition between labeled and unlabeled analytes for a limited number of binding sites on a capture molecule. In these assays, the target analyte is typically labeled with a detectable marker (e.g., a fluorescent molecule or enzyme). The labeled analyte competes with the unlabeled analyte in the sample for binding to the capture molecule. The amount of labeled analyte bound to the capture molecule is inversely proportional to the concentration of the unlabeled analyte present in the sample. The concentration of the unlabeled analyte can be quantified by measuring the signal generated by the labeled analyte.

[0008] Methods and apparatus for detecting the concentration of an analyte are disclosed in WO 2006 / 134546 A2. The method in WO 2006 / 134546 A2 is a competitive assay. In this method, magnetic trapping beads may have binding elements to bind the analyte to the sample. Furthermore, the analyte bound by the trapping beads may also bind to surface binding elements of a substrate.

[0009] Known immunoassays for vesicle analytes require labeling steps, and each analyte requires multiple antibodies to bind at different sites to distinguish nonspecific binding and background effects. Furthermore, such immunoassays typically do not meet the requirements of point-of-care testing (POCT). Additionally, known immunoassays cannot analyze untreated patient samples with low optical transparency, require washing steps prior to analysis, do not cover multiple biomarkers (e.g., vesicles, proteins, nucleic acids, cells) on a single readout platform, and employ expensive fluorescent labeling and complex readout systems to distinguish complex matrix effects. Moreover, known immunoassays do not allow for multiplex assays without sensor arrays or serial workflow solutions, driving up costs. Summary of the Invention

[0010] The disadvantages of known measurements are overcome by the method according to claim 1 and the apparatus according to claim 28. The dependent claims relate to particularly advantageous embodiments of the invention.

[0011] A first aspect of the present invention relates to a method for detecting the concentration of an analyte in a sample, the method comprising: A device with a channel is provided, the channel having a substrate having a surface binder element configured to bind with an analyte in the sample, and / or the substrate having a surface for testing the analyte; The measuring element, preferably a magnetic measuring element, is mixed into the sample, the measuring element having a capture binder element configured to bind to the analyte in the sample, and / or the measuring element having an element testing analyte configured to bind to the surface binder element; The sample flows through the channel; The measuring element is moved to the substrate by external force, preferably magnetic force; The measuring element is moved along / on the substrate, and the velocity of at least one measuring element is changed by interaction, wherein the interaction includes one or more of the following: (i) The interaction, preferably binding interaction, between at least one surface binder element of the substrate, the analyte, and at least one trapping binder element of the at least one measuring element; (ii) The interaction, preferably binding interaction, between the surface test analyte of the substrate and at least one of the trapping binder elements of the at least one measuring element; and / or (iii) The interaction, preferably binding interaction, between the element test analyte of the at least one measuring element and at least one of the surface binder elements of the substrate; and The concentration of the analyte is determined using a sensor unit by determining the velocity change of the measuring element along the channel, and / or by determining the loss of the measuring element along the channel, preferably by differential counting of the measuring element. Determine the binding kinetics, preferably binding constants, between the analyte and the trapping binder element and / or between the analyte and the surface binder element, wherein the binding kinetics are determined using the sensor unit by determining the velocity change of the measuring element along the channel.

[0012] By moving the assay element to and along the substrate / on the substrate, this method allows for the differentiation of assay elements bound to the analyte from those not bound to the analyte. Therefore, the concentration of the analyte in the sample can be determined. This method also allows for non-optical concentration determination, thus eliminating the need for pretreatment. The concentration of the analyte in opaque samples can be determined. Furthermore, concentrations can be determined over a wide range. Moreover, this invention allows for multiplex assays and / or heterogeneous assays, enabling the analysis of different types and / or species and / or categories of analytes in a sample, such as proteins, vesicle / vesicle material, cells / cell material, chemicals, DNA, RNA, and / or antibodies (serological).

[0013] Note that, in contrast, in WO 2006 / 134546 A2, the sample does not flow through the channel. Furthermore, WO 2006 / 134546 A2 does not disclose the external force (e.g., magnetic force) used to move the measuring element to the substrate, nor does it disclose the movement of the measuring element along / on the substrate, nor does it determine the analyte concentration by determining the velocity change and / or decrease of the measuring element along the channel, and / or by determining the loss of the measuring element along the channel. In contrast, WO 2006 / 134546 A2 determines the concentration by correlating the binding rate of the analyte to the substrate with a change in the magnetic field, recording the magnetic field change as magnetic (capture) beads carrying the analyte bind to the surface binder element.

[0014] In this invention, the sample may be a liquid sample or may include a liquid sample. The sample may be blood and / or urine or may include blood and / or urine. The analyte may be a vesicle, vesicle structure, one or more exosomes, mitochondria, etc. or may include vesicles, vesicle structures, one or more exosomes, mitochondria, etc. In some embodiments, the sample may contain or may contain more than one class and / or type of analyte, at least one of said analytes may be the target analyte whose concentration is to be detected. In other words, one or more classes and / or types of analytes may be analytes to determine and / or detect concentrations in the sample. The analyte may contain a ligand, such as an antigen. Different types and / or classes of analytes may have different ligands and / or antigens.

[0015] The apparatus may be an apparatus according to aspects of the invention or may include an apparatus according to aspects of the invention, as further detailed below.

[0016] Moving the measuring element along and / or on the substrate may include rolling the measuring element along and / or on the substrate.

[0017] When the assay elements are mixed in a sample, and when at least (or all) of the assay elements include one or more capture-binding elements, an analyte can bind to at least one of the assay elements via the corresponding capture-binding element, thereby forming at least one assay element with analyte binding. An assay element with analyte binding via the corresponding capture-binding element can be referred to as an analyte-bound assay element.

[0018] The capture binding element and / or surface binding element may each comprise, or may each comprise, one or more antibodies. The capture binding element may be the same antibody as the surface binding element or may include the same antibody as the surface binding element. The antibody may bind to an analyte and / or a ligand (e.g., antigen) that binds to the analyte. Different types and / or species of capture binding elements may include, for example, different antibodies, and / or may be configured to bind different species and / or types of analytes. Different types and / or species of surface binding elements may include, for example, different antibodies, and / or may be configured to bind different species and / or types of analytes. In some embodiments, it may be specified that the capture binding element of at least one, more, or all of the assay elements is of the same type and / or species as at least some, more, or all of the surface binding elements. Optionally or additionally, the capture binding element of at least one, more, or all of the assay elements may be matched with at least some, more, or all of the surface binding elements, such that the respective capture binding element and surface binding element form a matched antibody pair. In some embodiments, the capture binding element is a tetraspannome protein or includes tetraspannome proteins, such as CD81, CD63, and / or CD9. In some embodiments, the surface binding element is a tetraspannome protein or includes tetraspannome proteins, such as CD81, CD63, and / or CD9. In some embodiments, the capture binding element is an anti-Her2neu antibody and / or an anti-IL6 antibody or includes an anti-Her2neu antibody and / or an anti-IL6 antibody. In some embodiments, the analyte is Her2neu+ EV, IL6, and / or biotin or includes Her2neu+ EV, IL6, and / or biotin. In some embodiments, the substrate binding element is an anti-Her2neu antibody, an anti-IL6 antibody, and / or avidin or includes an anti-Her2neu antibody, an anti-IL6 antibody, and / or avidin.

[0019] An analytical element having a capture binder element can be a capture element or can correspond to a capture element. An analytical element having a capture binder element can be referred to as a "capture element". An analytical element having a capture binder element can capture analytes.

[0020] The interaction between the substrate's surface binder element, the analyte, and at least one trapping binder element can include the binding interaction between the analyte bound by the trapping binder element and the surface binder element. The same analyte can be bound to both the surface binder element and the trapping binder element simultaneously. The analyte can connect the surface binder element and the trapping binder element, or it can connect the surface binder element to the trapping binder element. The interaction between the substrate's surface binder element, the analyte, and at least one trapping binder element can include the interaction between the analyte and the trapping binder element, as well as another interaction between the analyte and the surface binder element.

[0021] The interaction between a surface-binding element and the analyte bound by the surface-binding element can include the binding interaction between the respective surface-binding element and the analyte. The capture-binding element and / or surface-binding element can be selected such that they can bind to a specific, selected, and / or desired analyte. In particular, if the sample contains more than one type and / or species of analyte, the capture-binding element and / or surface-binding element can be selected, for example, to bind to one of the analytes for which a concentration can be determined and / or tested, and / or whereby the analyte should be measured.

[0022] The interaction between the surface test analyte and the trapping binder element can include the binding interaction between the surface test analyte and the trapping binder element.

[0023] The interaction between the component test analyte and the surface binder component can include the bonding interaction between the component test analyte and the surface binder component.

[0024] Mixing the assay element into the sample can include or be equivalent to introducing the assay element into the sample. In some embodiments, multiple or all assay elements may be bound to the analyte in the sample, depending on the concentration of the analyte in the sample, the number of assay elements, the type and / or species of the analyte and / or the type and / or species of the capture-binding element of the corresponding assay element.

[0025] The binding kinetics (e.g., binding constant) between an analyte and a binding element (e.g., a trapping binding element or a surface binding element) can be determined by measuring the velocity variation of the measuring element along the channel. Greater velocity variations and / or losses result in stronger binding interactions and / or binding rates. Therefore, this method allows for the measurement and / or determination of binding interactions, such as binding constants, between different analytes. In some embodiments, this method can be used to analyze, determine, or tabulate the binding kinetics of analytes (e.g., analytes with unknown or only provisionally known binding kinetics). Furthermore, this method can be used to examine, validate, or verify known binding kinetics.

[0026] The measuring element may be or may include beads. The beads may be magnetic beads or may include magnetic beads. The measuring element and / or beads are magnetic. The measuring element and / or beads may be at least partially made of or contain magnetic material. In some embodiments, moving the magnetic measuring element and / or beads to the substrate includes pulling or attracting the magnetic measuring element and / or beads to the substrate. Alternatively or additionally, moving the magnetic measuring element and / or beads to the substrate may include pushing the magnetic measuring element and / or beads to the substrate. The external force may be a magnetic force provided by a magnetic field. In some embodiments, the magnetic field is a static magnetic field. Alternatively, the magnetic field may vary, for example, with time and / or along the channel. In some embodiments, the external force includes acoustic forces and / or gravity. In these cases, the measuring element need not be magnetic. The external force may be configured to move the measuring element toward the substrate. Additionally, the movement of the measuring element from the external force may govern Brownian motion and movement by diffusion. Thus, the measuring element can be reliably moved in the direction specified by the external force, for example, reliably pulled or attracted to the channel (wall). Due to external forces, the movement and / or attraction of the measuring element toward the channel (wall) can be substantially directional, rather than a random walk due to diffusion, for example. Therefore, compared to prior art such as WO 2006 / 134546 A2, the path traveled by the measuring element when moving toward the channel (wall) can be shorter and / or at least on average shorter. The contact and / or contact force between the measuring element and the substrate can be substantially caused by external forces. In some embodiments, the external forces are generated by an external force generator.

[0027] In some embodiments, multiple or all measuring elements are moved, pushed, and / or pulled to the channel and / or substrate. Preferably, all measuring elements are moved, pushed, and / or pulled to the channel and / or substrate. An external force can be configured to move all measuring elements to the channel and / or substrate. When one or more measuring elements move to the channel and / or substrate, the measuring elements can contact the channel and / or substrate.

[0028] It can be specified that the measuring element is detected by a sensor unit. In some embodiments, the measuring element comprises or is composed of a metal or alloy. Optionally or additionally, the measuring element may comprise or be composed of glass or plastic. It can be specified that the measuring element is coated or plated. For example, the measuring element may have a core comprising a first material (e.g., a suitable plastic, glass, or metal) and may be coated or plated with another material (e.g., a suitable plastic, glass, or metal). The plastic may be a polymer or may include a polymer. Optionally or additionally, the measuring element may comprise or be composed of biological materials. For example, the measuring element may have an internal material comprising a first material (e.g., a suitable polymer, glass, oil, protein, vesicle material, cell material, or metal) and may have an external material comprising a second material (e.g., a suitable polymer, glass, oil, protein, vesicle material, cell material, or metal).

[0029] The measuring element may be a bead or may include beads. For example, the beads may be metallic, and / or at least partially or entirely made of metal. It may be specified that the measuring element is spherical, but its form is not limited to this.

[0030] In some embodiments, the assay element comprises or is composed of cellular and / or vesicle material. Alternatively or additionally, the assay element may be or comprise stable droplets and / or cells. In some embodiments, the capture binder element is or corresponds to the outer surface of the assay element, for example, corresponding to the wall of the droplet and / or cell. Alternatively or additionally, the capture binder element may be or may correspond to the outer material of the assay element.

[0031] Flow through a channel, such as the flow of the measuring element and / or sample through the channel, can be laminar. However, the flow is not limited to this. Depending on, for example, the channel size, flow rate, viscosity of the flow and / or sample, smoothness of the channel surface, and / or other relevant parameters, the flow can be at least partially turbulent or can undergo a turbulent transition.

[0032] In some embodiments, multiple or all measuring elements move along / on the substrate. While moving along / on the substrate, a measuring element may roll along / on the substrate. Rolling on or along the substrate does not necessarily mean that the corresponding measuring element rolls over / along the entire length of the substrate. Rolling on or along the substrate may mean that the corresponding measuring element rolls over / along a portion of the substrate, which in some embodiments may be very short or minimal compared to the length of the substrate. Additionally or alternatively, rolling on or along the substrate may be facilitated by (but not necessarily only by) external forces and / or laminar flow. In some embodiments, rolling is facilitated, supported, assisted, and / or caused by external forces. It may be specified that the movement and / or speed of the measuring elements, particularly rolling on the substrate, depends on the external forces, for example, on the magnitude, amount, and / or power of the external forces. Alternatively or additionally, rolling may be facilitated, supported, assisted, and / or caused by flow through the channel. It can be specified that the movement and / or velocity of the measuring element, particularly its rolling on the substrate, depends on the flow, for example, on the flow velocity, such as the average flow velocity and / or flow rate. In some embodiments, the length by which the measuring element rolls on / along the substrate, or may roll, and / or the path of travel of the measuring element on / along the substrate, depends on, and / or is specified or determined by, external forces and / or the flow. The movement of the measuring element on or along the substrate may depend on, or may be specified or determined by, external forces and / or the flow.

[0033] When rolling on or along a substrate, the measuring element may at least partially contact the substrate. When rolling on or along a substrate, the measuring element may rotate while still at least partially contacting the substrate. When rolling on or along a substrate, the measuring element may rotate while moving on / along the substrate. The rolling motion may consist of translational and rotational motions, or may include both. The rolling motion may be, or may correspond to, a superposition of translational and rotational motions. Slippage of the measuring element may be negligible when it rolls on or along a substrate.

[0034] It may be specified that the speed of at least one, more, or all of the measuring elements is altered and / or reduced when moving or rolling on or along the substrate. Altering the speed of the measuring elements may include reducing the speed of at least one, more, or all of the measuring elements. It may be specified that the speed of measuring elements not bound to the analyte is not altered and / or reduced, or at least substantially not altered and / or reduced, when moving or rolling on or along the substrate.

[0035] Changing and / or reducing the speed may include having the measuring element adhered to or fixed to the substrate. Changing and / or reducing the speed of at least one of the measuring elements may include fixing the at least one measuring element, for example by binding a surface binder element to the analyte bound by a trap binder element. Alternatively or additionally, changing and / or reducing the speed of at least one of the measuring elements may include fixing the at least one measuring element, for example by binding the surface test analyte of the corresponding measuring element to a trap binder element of the substrate. Alternatively or additionally, changing and / or reducing the speed of at least one of the measuring elements may include fixing the at least one measuring element, for example by binding the element test analyte of the corresponding measuring element to a surface binder element of the substrate.

[0036] The analytes used in component testing can be of the same kind and / or type as the analytes in the sample. The analytes used in surface testing can be of the same kind and / or type as the analytes in the sample.

[0037] The sensor unit may include at least one sensing element. In a preferred embodiment, determining the concentration of the analyte includes determining the roll-off time between the first sensing element and the second sensing element. In some embodiments, the first sensing element is disposed upstream of the substrate. The second sensing element may be disposed downstream of the substrate and / or downstream of the first sensing element. It may be specified that the first sensing element is a magnetic field sensor and / or a magnetic sensor half-bridge, or includes a magnetic field sensor and / or a magnetic sensor half-bridge. The second sensing element may be a magnetic field sensor and / or a magnetic sensor half-bridge, or may include a magnetic field sensor and / or a magnetic sensor half-bridge.

[0038] Rolling time can be defined as the time required for a measuring element or a set of measuring elements to move or roll along or on the substrate between two defined points or regions on and / or upstream and / or downstream of the substrate (e.g., between two sensing elements (e.g., between the first sensing element and the second sensing element)), and / or between entering and leaving a region on the substrate.

[0039] The sensor unit and / or sensing element may be a device configured for impedance sensing and / or magnetoresistive sensing, or may include a device configured for impedance sensing and / or magnetoresistive sensing. The sensor unit and / or sensing element may detect and / or record a measuring element passing through, along, and / or through the sensor unit and / or sensing element. It may be specified that the sensor unit and / or sensing element measures and / or determines the size of the measuring element. Optionally or additionally, the sensor unit and / or sensing element may be a Coulter counter, or may include a Coulter counter.

[0040] In a preferred embodiment, the external force is a magnetic force generated by a static magnetic field, and the sensor unit may include at least one sensor configured to detect and / or determine the magnetic field. When the measuring element, particularly a magnetic measuring element and / or a magnetic bead, passes through the sensor unit, it may be specified that the change in the magnetic field detected and / or determined by the sensor unit allows for the detection of the measuring element (e.g., the magnetic measuring element) and / or the determination of its velocity.

[0041] In a preferred embodiment, the sample flows or is driven by a syringe, syringe pump, and / or metering pump. The syringe, syringe pump, and / or metering pump may be fluidly connected to a channel, for example, connected at or connected to the inlet of the channel. The syringe, syringe pump, and / or metering pump may apply force to the sample such that the sample velocity is substantially constant as it flows into or through the channel. Therefore, the velocity of the measuring element upstream and / or downstream of the substrate and / or at the inlet of the channel can be known. The velocity of the measuring element can be determined, detected, and / or measured by a sensor unit downstream of the substrate. The change and / or decrease in the velocity of the measuring element can then be determined by comparing, for example, the velocity applied at the inlet by the syringe, syringe pump, and / or metering pump with the velocity determined by the sensor unit.

[0042] In a preferred embodiment, determining the loss of the measuring elements includes differential counting of the measuring elements, or is performed by differential counting of the measuring elements. Differential counting can be performed by determining the difference between the number of measuring elements counted and / or recorded by two sensing elements. Alternatively or additionally, differential counting can be performed by determining the difference between the number of measuring elements counted and / or recorded by one sensing element and the known number of measuring elements before the sample with the measuring elements reaches the substrate. Loss of measuring elements may occur when one or more measuring elements are adhered to or fixed to the substrate. In some embodiments, providing a single sensing element for differential counting and / or a single sensing element is sufficient for differential counting because the number of measuring elements mixed into the sample may be known. The difference between the measuring elements mixed into the sample and the measuring elements recorded by the sensing elements can then be determined, making it possible to determine the number of measuring elements adhered to or fixed to the substrate upstream of the sensing elements.

[0043] Therefore, in some embodiments, non-optical determination of analyte concentration is possible. Thus, the method and apparatus according to the invention allow for determinations in which no sample pretreatment is required prior to the method. Furthermore, the sample can be opaque.

[0044] In a preferred embodiment, determining the speed change includes determining a first distribution function of the measuring element at a first position in the channel and a second distribution function of the measuring element at a second position in the channel. Alternatively or additionally, determining the rolling time may include determining the first distribution function of the measuring element at the first position in the channel and the second distribution function of the measuring element at the second position in the channel.

[0045] The first distribution function can be the distribution function of the time difference between the time point when the corresponding measuring element crosses the first position and the first reference time point. The second distribution function can be the distribution function of the time difference between the time point when the corresponding measuring element crosses the second position and the second reference time point.

[0046] The distribution function can be a histogram, a density function, and / or a cumulative function, or may include a histogram, a density function, and / or a cumulative function. The density function can be or may correspond to a standardized histogram. For example, the density function can be a standardized histogram or may include a standardized histogram such that the integral or sum over the entire histogram equals 1. The density function can be or may correspond to a probability density function.

[0047] The cumulative function can be a cumulative sum and / or a running sum, or may include cumulative sums and / or running sums. The cumulative function can be a cumulative distribution function, or may include a cumulative distribution function. In some implementations, the cumulative function can be an empirical cumulative distribution function (ECDF), or may include an empirical cumulative distribution function (ECDF).

[0048] The distribution function can be determined and / or measured by the sensing unit and / or by the sensing element. Alternatively or additionally, the distribution function can be determined optically (e.g., by using a camera device).

[0049] The first position can be the position of the first sensing element, and / or can coincide with the position of the first sensing element. Specifically, the first position can correspond to the position of the first sensing element in the channel. The second position can be the position of the second sensing element, and / or can coincide with the position of the second sensing element. Specifically, the second position can correspond to the position of the second sensing element in the channel.

[0050] In a preferred embodiment, the first distribution function includes a density function or histogram, and the second distribution function includes a density function or histogram, and the speed change is determined by the time difference between the peak of the second distribution function and the peak of the first distribution function. The peak of the first distribution function may be, or may correspond to, the maximum value of the first distribution function. The peak of the second distribution function may be, or may correspond to, the maximum value of the second distribution function. Since the peaks can be easily identified in some embodiments, the change in speed (and / or rolling time) can be easily determined.

[0051] In a preferred embodiment, the first distribution function includes a cumulative function, and the second distribution function includes a cumulative function, and the rate change is determined by the time difference between the inflection point of the second distribution function and the inflection point of the first distribution function. In some embodiments, the cumulative function can be determined more easily than a histogram or its corresponding density function. In some embodiments, the cumulative function can be determined more accurately than its corresponding density function.

[0052] In a preferred embodiment, the velocity change is determined by a distance metric between the second distribution function and the first distribution function. Alternatively or additionally, the velocity change can be determined by the shape change of the second distribution function relative to the first distribution function. In some embodiments, the distance metric and / or shape change can be determined more easily and / or more accurately than calculating the time difference between peaks and / or inflection points. For example, when the distribution has more than one peak and / or more than one inflection point, the distance metric and / or shape change can be more accurate and / or less arbitrary.

[0053] The distance metric can be the Kolmogorov-Smirnov test, Wasserstein metric, divergence metric, entropy metric (e.g., Kullback-Leibler divergence), or some other suitable function, such as, but not limited to, f-divergence, Jenson-Shannon divergence, Bhattacharyya distance, etc.

[0054] Shape changes can be determined by fitting the first and second distribution functions to the same candidate distribution function and comparing their respective fitting parameters. The candidate distribution function can be a normal distribution function, a gamma distribution function, a log-normal distribution function, or a Weibull distribution function, but is not limited to these. In some embodiments, the distribution function can be a distribution function with a support set [0, x), where x is a positive number. In some embodiments, the distribution function can be a distribution function with a support set [0, ∞).

[0055] In the preferred embodiment, the first reference time point is equivalent to the second reference time point. Therefore, the time difference can be accurately determined because both distributions are functions of time relative to the same reference point or zero point.

[0056] In a preferred embodiment, the first reference time point is equivalent to the time point when the measuring element first crosses the first position. Alternatively or additionally, the second reference time point may be equivalent to the time point when the measuring element first crosses the second position. This allows the first and second distribution functions to be shifted to the left, so that they start from the origin. Therefore, determining distance measurements and / or shape changes can be facilitated and / or made more accurate. For example, shifting the first and / or second distribution functions to the left may result in increased overlap between the first and second distribution functions.

[0057] In a preferred embodiment, determining the concentration of the analyte includes optical detection. Optical detection may include capturing the movement and / or position of a spatially and / or temporally resolved measuring element within a field of view. By determining the position and / or movement of the measuring element on the substrate, rolling time and / or differential counts can be determined optically. Optical detection may be performed by one or more optical devices, such as one or more cameras. Each optical device and / or camera may have a field of view. In some embodiments, more than one optical device and / or more than one camera is provided, for example, to have a larger total field of view at sufficient resolution. The fields of view of at least two optical devices and / or cameras may at least partially overlap. In some embodiments, the fields of view of at least two optical devices and / or cameras do not overlap. It may be specified that a first optical device and / or camera is arranged such that its field of view covers an area upstream of the substrate, and a second optical device and / or camera is arranged such that its field of view covers an area downstream of the substrate. Optical detection may detect one or more of the color, size, luminescence, and particle size of the measuring element. Therefore, in some embodiments, multiple determinations are possible.

[0058] In a preferred embodiment, the external force (e.g., magnetic force provided by a magnetic field) and the sample flow rate are adjusted such that at least one of the measuring elements moves, rolls, and / or rolls along / on the substrate. For example, in some embodiments, when moving toward the substrate, the measuring element contacts the channel and / or substrate at substantially similar locations or at least in a defined area on the channel and / or substrate. Thus, for example, the length and / or distance of rolling on the substrate can be fixed, or at least more precisely specified. In some embodiments, the flow rate is selected such that the measuring element slows down only when moving or rolling along / on the substrate, but is not stuck or fixed.

[0059] In a preferred embodiment, an external force (e.g., magnetic force provided by a magnetic field) is adjusted to push the assay element away from the substrate and / or sort it. In some embodiments, the external force is adjusted to push the assay element away from the substrate and / or sort it, preferably after the analyte concentration has been determined. Thus, the channel and / or substrate can be prepared for another analysis or assay, and / or for assay elements to be further analyzed using different methods. For example, an assay element adhered to the substrate can be released from the substrate, and then the assay element can be used, for example, for PCR (polymerase chain reaction). Optionally or additionally, the temperature can be increased to minimize nonspecific binding on the substrate or to achieve dissociation of the assay element. In particular, when the binding is DNA-DNA, RNA-DNA, or DNA-PNA binding, the strength of the binding can be altered and / or reduced by increasing the temperature. In some embodiments, the temperature can be increased to be close to or greater than the corresponding melting temperature of the binding, for example, the corresponding melting temperature of the DNA-DNA binding, RNA-DNA binding, and / or DNA-PNA binding.

[0060] In a preferred embodiment, a laminated flow flows between the sample and the substrate within the channel, for example, to separate the sample from the substrate. The measuring element can be moved from the sample to the substrate by an external force (e.g., magnetic force generated by a magnetic field) through the laminated flow. Viewed in a longitudinal section of the channel, the height of the laminated flow can be greater than the maximum size of the measuring element, e.g., its diameter. In some embodiments, more than one laminated flow flows on the substrate, wherein the laminated flows can substantially overlap each other, e.g., in a direction perpendicular to the substrate surface. The laminated flow can prevent debris from clogging or hindering the measuring element from moving or rolling along or on the substrate. The laminated flow can, for example, minimize interference from hematocrit. In some embodiments, the laminated flow can be a sheath flow or may include a sheath flow.

[0061] In a preferred embodiment, at least one, at least two, more, or all of the measuring elements each have at least two different trapping binder elements. The different trapping binder elements can be configured to bind to different analytes. Therefore, the method allows for multiple determinations. At least two, more, or all of the measuring elements can have different sizes, different magnetic moments, different acoustic properties, different electrical properties, and / or different optical properties. It can be specified that the measuring elements can be distinguished or grouped based on their different sizes, different magnetic moments, different acoustic properties, different electrical properties, and / or different optical properties.

[0062] Optical properties may include color or markings, such that, for example, the measuring element may be a colored or marked measuring element, but are not limited thereto. Acoustic properties may include compressibility and / or density, such as, for example, the compressibility and / or density of the measuring element, but are not limited thereto. Electrical properties may include impedance and / or charge, such as, for example, the impedance and / or charge of the measuring element, but are not limited thereto.

[0063] Alternatively or additionally, different capture-binding elements can be configured to bind to the same analyte. For example, the analyte may be a vesicle with different proteins or may include vesicles with different proteins, such as, for example, CD9, CD63, or CD81 in the case of extracellular vesicles. These proteins can bind to different capture-binding elements. Therefore, binding of the analyte to the assay element can be facilitated. Alternatively or additionally, different capture-binding elements may contain different antibodies, which can be configured to bind different proteins of the same analyte.

[0064] In a preferred embodiment, at least two of the measuring elements have different sizes and different trapping binder elements, wherein the different trapping binder elements can be configured to bind to different analytes. Alternatively or additionally, at least two of the measuring elements may have different magnetic moments and different trapping binder elements, wherein the different trapping binder elements can be configured to bind to different analytes. Alternatively or additionally, at least two of the measuring elements may have different optical properties and different trapping binder elements, wherein the different trapping binder elements can be configured to bind to different analytes. Alternatively or additionally, at least two of the measuring elements may have different acoustic properties and different trapping binder elements, wherein the different trapping binder elements can be configured to bind to different analytes. Alternatively or additionally, at least two of the measuring elements may have different electrical properties and different trapping binder elements, wherein the different trapping binder elements can be configured to bind to different analytes. In some embodiments, the measuring element may be specified to include at least two sets of measuring elements, and each set of measuring elements may have the same trapping binder element. At least two of the measuring elements, and / or at least two groups of measuring elements, may differ in at least one or more of size, color, luminescence, particle size, and magnetic moment. Therefore, this method allows for multiple measurements.

[0065] In a preferred embodiment, the substrate has at least two different surface binder elements, wherein the different surface binder elements can be configured to bind to different analytes. In some embodiments, the different surface binder elements may be specified to be grouped into homogeneous groups, preferably separated by a sensing element. Therefore, this method allows for multiple determinations.

[0066] In some embodiments, the capture binder element of at least one or all of the measuring elements is of the same type and / or kind as at least one or all of the surface binder elements.

[0067] This method allows for multiplex and / or heterogeneous assays. It can be used to analyze different types and / or species and / or categories of analytes in a sample, such as proteins, vesicles / vesicle material, cells / cell material, chemicals, DNA, RNA, antibodies (serological), and / or to determine the concentrations of multiple analytes in the sample. Different assay binding elements can differ in type and / or species, for example, they may include or may be different antibodies. Different surface binding elements can differ in type and / or species, for example, they may include or may be different antibodies. In some embodiments, the different capture binding elements differ in the number of capture binding elements disposed on the surface of the respective assay element.

[0068] It can be specified that the orientation and / or rolling direction of the measuring element changes as it moves or rolls on or along the substrate. Therefore, when moving or rolling along or on the substrate, a larger portion of the measuring element's surface comes into contact with the surface binder element, promoting the binding of the surface binder element to the analyte bound to the rolling measuring element. This can improve the accuracy of the method because, for example, measuring elements bound to the analyte can be better distinguished from those not bound to the analyte.

[0069] Alternatively or additionally, it may be specified that the orientation and / or rolling direction of the measuring element changes as it moves or rolls downstream of the substrate or the sensing unit. For example, the orientation and / or rolling direction can be changed by a pattern. The pattern may be a mechanical pattern or may include a mechanical pattern. A mechanical pattern can guide the measuring element through mechanical interactions (e.g., through one or more guide elements). Alternatively or additionally, the pattern may be a magnetic pattern or may include a magnetic pattern. A magnetic pattern can guide the measuring element through a magnetic field and / or a magnetic force acting on the measuring element.

[0070] Optionally or additionally, an external force may alter the orientation and / or rolling direction of the measuring element. It may be specified that the external force can be controlled such that the orientation and / or rolling direction of the measuring element can be altered and / or varied.

[0071] In a preferred embodiment, a second external force acts on the measuring element as it moves downstream of the substrate and / or downstream of the sensing unit. This second external force can sort the measuring elements. In some embodiments, the measuring elements can be sorted or grouped according to their different characteristics, such as, for example, their different magnetic moments, different sizes, different acoustic properties, different electrical properties, and / or different optical properties. In some embodiments, the second external force can singulate at least one, more, or all of the measuring elements. The second external force can be a second magnetic force, but is not limited to this. The second external force can differ from the external force, for example, in amplitude, power, and / or frequency. It can be specified that the second external force is controlled and / or can vary, for example, over time and / or locally. Thus, the sorting and / or singulation of the measuring elements can be controlled. Sorting and / or singulating the measuring elements can be useful for reusing the device and / or the measuring elements.

[0072] In some implementations, a second external force can change the orientation and / or rolling direction of the measuring element as it moves or rolls on or downstream of the substrate or the sensing unit.

[0073] A second aspect of the invention relates to an apparatus for detecting the concentration of an analyte in a sample, the apparatus comprising a channel having a substrate having a surface binder element and / or having a surface test analyte. The channel is configured such that a sample containing the analyte and a measuring element can flow through it, the measuring element having a capture binder element and / or having an element test analyte. The apparatus is configured such that, as the sample containing the analyte and the measuring element flows through the channel, an external force moves the measuring element to the substrate. The channel is configured such that, as the sample flows through the channel, the measuring element moves along / on the substrate. The apparatus is further configured such that, as the measuring element moves along / on the substrate, the velocity of at least one measuring element is changed by an interaction, the interaction including one or more of the following: (i) The interaction, preferably binding interaction, between at least one surface binder element of the substrate, the analyte, and at least one trapping binder element of the at least one measuring element; (ii) The interaction, preferably binding interaction, between the surface test analyte of the substrate and at least one trapping binder element of the at least one measuring element; and / or (iii) The interaction, preferably a bonding interaction, between the test analyte of the at least one measuring element and at least one of the surface binder elements of the substrate.

[0074] The apparatus further includes a sensor unit configured to determine the velocity variation of the measuring element along the channel, and / or determine the loss of the measuring element along the channel, preferably by differential counting of the measuring element. Optionally or additionally, the apparatus is configured to determine the binding kinetics, preferably the binding constant, between the analyte and the trapping binder element and / or between the analyte and the surface binder element.

[0075] The device can also be configured to determine the concentration and / or binding kinetics of the analyte by a determined rate change.

[0076] It can be specified that, as the measuring elements flow through the channel, multiple or all of the measuring elements roll along / on the substrate. "Moving" the measuring elements along / on the substrate can include rolling the measuring elements along / on the substrate.

[0077] The apparatus can be configured to perform the method according to the first aspect of the invention. The apparatus can be used to determine the concentration of an analyte in a sample. The apparatus can be used to determine binding kinetics. The apparatus may have one, more, or all of the advantages of the method of the invention described herein.

[0078] In some embodiments, the device may include a sample. In other embodiments, the device may not include a sample. In some embodiments, the device may include a measuring element. In other embodiments, the device may not include a measuring element.

[0079] In some embodiments, the device includes an external force generating device configured to generate an external force. The external force generating device may be a magnetic field generating device configured to generate a magnetic field, or may include a magnetic field generating device configured to generate a magnetic field, such that the external force may be magnetic. The external force (e.g., magnetic force) may be configured to move a measuring element to a substrate. The measuring element may be a magnetic bead, or may include a magnetic bead, and / or may be magnetic.

[0080] The sensor unit may include at least one sensing element. In a preferred embodiment, the sensor unit includes a first sensing element and a second sensing element, wherein the sensor unit may be configured to determine the concentration of the analyte by determining the roll-off time between the first sensing element and the second sensing element. In some embodiments, the first sensing element is disposed upstream of the substrate. The second sensing element may be disposed downstream of the first sensing element and / or the substrate. The first sensing element may be a magnetic field sensor and / or a magnetic sensor half-bridge, or may include a magnetic field sensor and / or a magnetic sensor half-bridge. The second sensing element may be a magnetic field sensor and / or a magnetic sensor half-bridge, or may include a magnetic field sensor and / or a magnetic sensor half-bridge.

[0081] In a preferred embodiment, the external force generating device is a magnetic field generating device, such as a permanent magnet. The external force can be magnetic. The magnetic field generating device can generate a static magnetic field. The sensor unit can include at least one sensor configured to detect and / or determine the magnetic field and / or changes in the magnetic field. When the measuring element, particularly a magnetic measuring element and / or a magnetic bead, passes through the sensor unit, it can be specified that the change in the magnetic field detected and / or determined by the sensor unit allows the measuring element (e.g., a magnetic measuring element) to be detected and / or its velocity to be determined.

[0082] In a preferred embodiment, the device and / or wherein the sensing unit is configured to determine velocity changes by determining a first distribution function of the measuring element at a first position in the channel and a second distribution function of the measuring element at a second position in the channel. The first distribution function may be a distribution function of the time difference between the time point at which the corresponding measuring element crosses the first position and a first reference time point. The second distribution function may be a distribution function of the time difference between the time point at which the corresponding measuring element crosses the second position and a second reference time point.

[0083] The first position can be the position of the first sensing element, such as the position or location of the first sensing element in the channel. The second position can be the position of the second sensing element, such as the position or location of the first sensing element in the channel.

[0084] In a preferred embodiment, the first distribution function includes a density function or a histogram, and the second distribution function includes a density function or a histogram. The velocity change can be determined by the time difference between the peak (preferably the maximum value) of the second distribution function and the peak (preferably the maximum value) of the first distribution function.

[0085] In a preferred embodiment, the first distribution function includes a cumulative function, and the second distribution function also includes a cumulative function. The velocity change can be determined by the time difference between the inflection point of the second distribution function and the inflection point of the first distribution function.

[0086] In a preferred embodiment, the velocity change is determined by a distance metric between the second distribution function and the first distribution function, and / or by a shape change of the second distribution function compared to the first distribution function.

[0087] In a preferred embodiment, the sensor unit is an optical device or includes an optical device. The optical device can be configured to determine the concentration of the analyte by capturing the motion and / or position of a measuring element spatially and / or temporally resolved within a field of view. The optical device may include one or more cameras. Each optical device and / or camera may have a field of view. In some embodiments, more than one optical device and / or more than one camera is provided, for example, to have a larger total field of view at sufficient resolution. The fields of view of at least two optical devices and / or cameras may at least partially overlap. In some embodiments, the fields of view of at least two optical devices and / or cameras do not overlap. It may be specified that a first optical device and / or camera is arranged upstream of the substrate and / or such that its field of view covers an upstream region of the substrate, and a second optical device and / or camera is arranged downstream of the substrate and / or such that its field of view covers a downstream region of the substrate. The optical device can be configured to detect one or more of the color, size, luminescence, and particle size of the measuring element. Thus, in some embodiments, multiplex measurements are possible.

[0088] In a preferred embodiment, the substrate has at least two different surface binder elements, wherein the different surface binder elements can be configured to bind to different analytes. It can be specified that the different surface binder elements are grouped into homogeneous groups separated by sensing elements.

[0089] Alternatively or additionally, the substrate may have at least two different surface binding elements, wherein the different surface binding elements can be configured to bind to the same analyte. For example, the analyte may be or may contain vesicles with different proteins, such as, for example, in the case of extracellular vesicles, CD9, CD63, or CD81. These proteins can bind to different surface binding elements. Thus, the binding of the analyte to the substrate can be facilitated. Alternatively or additionally, the different substrate binding elements may contain different antibodies, which can be configured to bind different proteins of the same analyte.

[0090] In a preferred embodiment, the device is configured to change the orientation and / or rolling direction of the measuring element while the measuring element is rolling along / on the substrate or downstream of the substrate or downstream of the sensor unit and / or before the measuring element rolls along / on / on the substrate or downstream of the substrate or downstream of the sensor unit.

[0091] In a preferred embodiment, the channel and / or substrate have mechanical and / or magnetic patterns. The mechanical and / or magnetic patterns may be mechanical and / or magnetic herringbone patterns or may include mechanical and / or magnetic herringbone patterns. The mechanical and / or magnetic patterns may be mechanical and / or magnetic structures or may include mechanical and / or magnetic structures. The mechanical and / or magnetic patterns can be configured to change the orientation and / or direction of movement or rolling of the measuring element as it moves or rolls along / on the substrate. Therefore, the likelihood or opportunity for the analyte bound to the capture binder element to contact and / or interact with the surface binder element is increased.

[0092] In a preferred embodiment, the channel may be tapered. When the channel is tapered, its cross-section may vary along its length. As the cross-section increases, the flow velocity may decrease. The channel may be tapered such that its cross-section increases along the flow direction. When the channel is tapered, the driving force exerted by the flow on the sensing element may decrease, and the velocity of the sensing element may further decrease. In some embodiments, the likelihood of the sensing element being fixed or stuck may be increased. In some embodiments, the tapering of the channel guides and / or orients the sensing element to the sensing unit.

[0093] In a preferred embodiment, the device includes a second external force generating device configured to generate a second external force. The second external force generating device is arranged downstream of the sensor unit. Alternatively or additionally, the second external force generating device may be arranged along a substrate. In some embodiments, the second external force generating device is or includes a second magnetic field generating device, and the second external force is magnetic. The second magnetic field generating device may be or may include a permanent magnet. The second external force generating device may be configured to generate a second external force with varying amplitude and / or spatial orientation. The second external force may vary over time. The second external force generating device may be specified as a sorting unit for sorting measurement elements. For example, the second external force may be used to sort measurement elements, such as those downstream of the sensor unit. Through sorting, the measurement elements can be grouped into different groups. The different groups of measurement elements can be used for further analysis or diagnostics.

[0094] The second external force can be a second magnetic force, but is not limited to this. The second external force can be different from the external force, for example, it can differ in amplitude, power, and / or frequency. It can be specified that the second external force is controlled and / or can vary, for example, with time and / or locally.

[0095] Optionally or additionally, the second external force generating device and / or the second external force can be configured to change the orientation and / or rolling direction of the measuring element. Optionally or additionally, the second external force generating device and / or the second external force can be configured to simplify one or more of the measuring elements.

[0096] The second external force generating device may include an external force generating device. Alternatively, the external force generating device may include a second external force generating device. Attached Figure Description

[0097] The invention is further disclosed and described in detail with reference to the following figures: Figure 1 A schematic diagram of an apparatus according to an embodiment of the present invention; Figure 2 Another schematic diagram of an apparatus according to an embodiment of the present invention; Figure 3 : Schematic diagram of a measuring element with a binder-capturing element; Figure 4 A schematic diagram of the measuring element and substrate according to another embodiment of the present invention; Figure 5 A schematic diagram of the measuring element and substrate according to another embodiment of the present invention; Figure 6 A flowchart illustrating a method according to an embodiment of the present invention is provided. Figure 7 : Another flowchart illustrating a method according to an embodiment of the present invention; Figure 8 At the first time t1, a schematic diagram of the apparatus according to an embodiment of the present invention is shown; Figure 9 : At the second time t2 Figure 8 Implementation plan; Figure 10 : Figure 8 and Figure 9 An exemplary output of the sensor unit in the implementation scheme; Figure 11 : Figures 8 to 10 A comparison of the proposed implementation plan with conventional measurements; Figure 12 : An exemplary output of a sensor unit for a single measuring element; Figure 13 Example of a distribution function; Figure 14 Another example of a distribution function; Figure 15 Another example of a distribution function; Figure 16Example curve of the shape factor of the distribution function as a function of analyte concentration.

[0098] Figure 17 A schematic diagram of an apparatus according to another embodiment of the present invention; Figure 18 : Figure 17 A comparison of the proposed implementation plan with conventional measurements; Figure 19 A schematic diagram of an apparatus according to another embodiment of the present invention; Figure 20 : Figure 19 An exemplary output of the sensor unit in the implementation scheme; Figure 21 : Figure 19 and Figure 20 A comparison of the proposed implementation plan with conventional measurements; Figure 22 A schematic diagram of an apparatus having sheath flow according to an embodiment of the present invention; Figure 23 : A schematic diagram of an apparatus according to an embodiment of the present invention, having multiple measurement capabilities; Figure 24 : A schematic diagram of an apparatus according to another embodiment of the present invention, having multiple measurement capabilities; Figure 25 A schematic diagram of an apparatus according to another embodiment of the present invention, having multiple measurement capabilities; Figure 26 A schematic diagram of an apparatus according to an embodiment of the present invention, the apparatus having a pattern for changing the direction and / or orientation of the measuring element; Figure 27 A schematic diagram of an apparatus according to another embodiment of the present invention, the apparatus having a pattern for changing the direction and / or orientation of the measuring element; Figure 28 A schematic diagram of an apparatus according to another embodiment of the present invention, the apparatus having a pattern for changing the direction and / or orientation of the measuring element; Figure 29 A schematic diagram of an apparatus according to an embodiment of the present invention, the apparatus having a tapered channel; and Figure 30a : An exemplary analyte concentration determined as a function of the rate change of the measuring element by the method according to the invention; Figure 30b Exemplary analyte concentrations determined by the method according to the invention as a function of the proportion of the measuring element that is fixed and / or adhered; Figure 31aComparison of exemplary analyte concentrations as a function of the proportion of fixed and / or adhered measuring elements, determined by different methods according to the present invention; Figure 31b : Comparison of exemplary analyte concentrations as a function of the proportion of fixed and / or adhered measuring elements, determined by different methods according to the invention; and Figure 32 Example curve showing the rate of decrease as a function of analyte concentration. Detailed Implementation

[0099] It should be understood that the general description above and the description below are merely exemplary and illustrative, and do not limit the methods and apparatus described herein. In this application, the use of the singular may include the plural unless specifically stated otherwise. Furthermore, where applicable or unless otherwise stated, the use of "or" means "and / or". Those skilled in the art will recognize that the following description is merely illustrative and not intended to be limiting in any way. Other embodiments will readily conceive of those who benefit from this disclosure. Various embodiments of the exemplary embodiments, as illustrated in the accompanying drawings, will now be described in detail. In the drawings, the same reference numerals may denote the same objects.

[0100] Figure 1 and Figure 2 An exemplary embodiment of the apparatus 100 according to the present invention is shown. The apparatus 100 can be considered a testing device for performing a determination. The apparatus 100 can be used to determine the concentration of analyte 10 in sample 20. The apparatus 100 can be configured to perform the method according to the present invention.

[0101] Sample 20 may include a liquid or may be a liquid sample 20. For example, sample 20 may be blood or urine or may include blood or urine. Sample 20 may contain one or more types and / or species of analyte 10 to be analyzed. Analyte 10 may be interleukin-6 (IL-6) or may include interleukin-6 (IL-6). Analyte 10 may be a vesicle or may include vesicles. Analyte 10 may be a vesicle structure, one or more exosomes, mitochondria, etc. or may include a vesicle structure, one or more exosomes, mitochondria, etc. However, other and / or different analytes, and / or types and / or species of analyte 10 may also be analyzed and / or detected.

[0102] The apparatus 100 includes a channel 40. A sample 20 having an analyte 10 and a measuring element 30 having at least one analyte bond, and / or a sample 20 having a measuring element 30, can flow through the channel 40. In particular, a flow 110 of the sample 20 (in which the measuring element 30 is mixed) can flow through the channel 40. The flow 110 can be laminar flow. The measuring element 30 can be magnetic. The measuring element 30 can be a magnetic bead or may include magnetic beads.

[0103] Channel 40 may be a microfluidic channel or may include a microfluidic channel. Channel 40 may have an inlet and an outlet for allowing sample 20 to flow through channel 40. Channel 40 may have a transverse diameter in the zy plane (e.g., Figure 1 and Figure 2 Fluid 110 can flow through this transverse diameter. The transverse diameter can be substantially rectangular, elliptical, circular, or have any other suitable shape. For example, the flow direction through channel 40 can be the x-direction. In some embodiments, channel 40 can be arranged and / or oriented such that its length direction extends substantially perpendicular to gravity. For example, gravity can act along the negative z-direction.

[0104] Channel 40 has a base 42. The base 42 can be disposed on the inner wall of channel 40, or can be part of the inner wall of channel 40. The base 42 can be disposed at the bottom of channel 40. The base 42 can extend through channel 40 in the flow direction (e.g., in…). Figure 1 and Figure 2 (in the x direction). The substrate 42 may be a plane or a surface, or may include a plane or a surface, and / or may extend in a direction perpendicular to its length direction (e.g., in the x direction). Figure 1 and Figure 2 (in the y-direction). The base 42 can be arranged such that the flow 110 flows on / above the base 42. Gravity can act in the negative z-direction (see, for example, Figure 1 and Figure 2 Furthermore, the base 42 can be arranged such that it supports the flow 110. However, different arrangements and / or orientations of the channel 40 and / or the base 42 are possible.

[0105] The substrate 42 includes a surface binder element 44. The surface binder element 44 is disposed on the surface of the substrate 42, along which the sample 20 flows, and / or the measuring element 30 moves or rolls along or on the surface of the substrate 42. The surface binder element 44 may extend from the substrate 42. The surface binder element 44 may bind to the analyte 10, and / or the analyte 10 may bind to the surface binder element 44.

[0106] Surface binding element 44 may be an antibody for binding analyte 10 or may include an antibody for binding analyte 10. Analyte 10 may be a ligand and / or an antigen or may include a ligand and / or an antigen to bind to surface binding element 44 and / or a corresponding antibody. Surface binding elements 44 may be selected such that they can bind to a specific, selected, and / or desired analyte 10. Capture binding elements 35, 36, and 37 may have the same antibody as surface binding elements 44, 45, and 46. In some embodiments, surface binding elements 44, 45, and 46 are tetraspan proteins or include tetraspan proteins such as CD81, CD63, and / or CD9. In some embodiments, capture binding elements 35, 36, and 37 are or contain anti-Her2neu antibodies and / or anti-IL6 antibodies. In some embodiments, analyte 10 is Her2neu+ EV, IL6, and / or biotin or includes Her2neu+ EV, IL6, and / or biotin. In some embodiments, the base binding element 42 is an anti-Her2neu antibody, an anti-IL6 antibody, and / or avidin, or includes an anti-Her2neu antibody, an anti-IL6 antibody, and / or avidin.

[0107] The device 100 may also include an external force generating device 120. The external force generating device 120 may be a magnetic field generating device 120 or may include a magnetic field generating device 120. The external force generating device 120 may be configured to generate an external force 60. The external force 60 may be a magnetic force (e.g., generated by a magnetic field) and / or an acoustic force or may include a magnetic force (e.g., generated by a magnetic field) and / or an acoustic force. The magnetic field generating device may be configured to generate a magnetic field and / or a magnetic force. In some embodiments, the external force generating device 120 and / or the magnetic field generating device are arranged at the channel 40. The magnetic field generating device may be a magnet or may include a magnet, such as a permanent magnet. For example, in Figure 1 In the implementation plan, the magnetic field generating device includes a magnetic north pole and a magnetic south pole. It can be specified that the magnetic field is a static magnetic field.

[0108] Alternatively or additionally, the magnetic field generating device may be an electromagnet or may include an electromagnet. However, the magnetic field generating device is not limited to this. In some embodiments, for example, when the magnetic field generating device is an electromagnet or includes an electromagnet, it may be specified that the magnetic field is variable, such as local or time-varying.

[0109] When the measuring element 30 is magnetic and / or includes magnetic beads, the magnetic field and / or magnetic field generating device can be configured to move the measuring element 30 to the substrate 42, for example, by attracting the measuring element 30.

[0110] In some embodiments, device 100 includes more than one or more external force generating devices 120 and / or magnetic field generating devices, such as Figure 2As shown in the embodiments. In some embodiments, when the device 100 includes more than one external force generating device 120 and / or more than one magnetic field generating device, the external force 60 (e.g., magnetic force) generated by each external force generating device 120 is different in, for example, intensity and / or orientation.

[0111] It may be specified that the external force 60 (e.g., a magnetic field and / or magnetic force) is altered or reversed. In some cases, the external force 60 is altered or reversed to release the measuring element 30, which is adhered to and / or fixed, from the substrate 42. In some embodiments, the channel 40 and / or the substrate 42 are released from the adhered and / or fixed measuring element 30 when the analyte concentration has been determined and / or after the analyte concentration analysis has been completed.

[0112] In some implementations, for example, in Figure 1 and Figure 2 In those cases, an external force generating device 120 (e.g., a magnetic field generating device 120) is arranged below the substrate 42, for example, below the substrate 42 in the z-direction. The external force generating device 120 can apply a pulling force to the measuring element 30 flowing within the channel 40 and / or the measuring element 30 above the substrate 42. Therefore, the measuring element 30 flowing within the channel 40 can be moved to or is being moved to the substrate 42.

[0113] Alternatively or additionally, in some embodiments, one or more of the external force generating devices 120 (e.g., one or more magnetic field generating devices 120) are arranged above the substrate 42, for example, at or above the upper sidewall of the channel 40, or may be arranged opposite to the substrate 42. The external force generating devices 120 may apply thrust to the measuring elements 30 flowing within the channel 40 and / or above the substrate 42. Thus, the measuring elements 30 flowing within the channel 40 may be moved to or may be moving to the substrate 42. Alternatively or additionally, the external force generating devices 120 may apply thrust to the measuring elements 30 such that they may be sorted.

[0114] In some embodiments, the external force 60 is gravity or includes gravity. In some embodiments, it may be specified that the external force 60 is superimposed with gravity, and / or includes gravity in addition to another force. If the external force 60 is gravity or includes gravity, then in some embodiments, the device 100 does not include the external force generating device 120. However, in some other embodiments, even when the external force 60 is gravity or includes gravity, the device 100 includes the external force generating device 120. In some embodiments, the flow rate and / or measuring element 30 is selected and / or configured, for example, the measuring element 30 may contain suitable materials and / or may have suitable dimensions, such that gravity is sufficient to move the measuring element 30 toward the substrate 42. In some other embodiments, gravity is insufficient to cause or provide said movement of the measuring element 30.

[0115] It can be specified that the external force 60 (e.g., a magnetic field and / or magnetic force) is adjusted, selected, or changed in conjunction with the flow velocity of the flow 110 such that the measuring element 30 can move or roll along or on the substrate 42. Specifically, the flow velocity of the flow 110 and the external force 60 can be adjusted, selected, or changed such that all or selected or specific portions of the measuring element 30 can move or roll along or on the substrate 42. The flow velocity of the flow 110 and the magnetic field 60 can be adjusted, selected, or changed such that all or selected or specific portions of the measuring element 30 can contact the substrate 42 at or near a designated point along the flow direction (e.g., along the x-direction in the figure).

[0116] In some embodiments, the flow rate of flow 110 is selected such that, when moving or rolling along or on substrate 42, the analyte-bound measuring element 30 is only slowed down, but not stuck or fixed. In some embodiments, for example, the flow rate of flow 110 is controlled or varied according to the size of the measuring element 30 and / or the strength of the binding interaction.

[0117] The external force 60 and / or the external force generating device 120 may be configured and / or arranged such that some, most or all of the measuring elements 30 can contact the inner wall of the channel 40 and can move or roll along / on the inner wall before moving or rolling onto the base 42.

[0118] Device 100 includes sensor unit 70 ( Figure 1 and Figure 2 (Not shown in the image). See below for reference. Figures 8 to 29 An exemplary embodiment of the sensor unit 70 is described. For example, Figure 1 and Figure 2 The sensor unit 70 of the embodiment (not shown in those figures) may correspond to or be equivalent to the following and / or the sensor unit 70 described below. Figure 8 , Figure 10 , Figure 11 , Figure 19 , Figure 26 , Figure 27 , Figure 28 or Figure 29 One or more of the sensor units 70 shown.

[0119] Apparatus 100 may include a control unit (not shown). The control unit may be a computer unit or may include a computer unit. The control unit may be configured to control the flow rate of the external force generating device 120, the external force 60, and / or the flow 110. The control unit may be configured to determine the concentration of analyte 10 in sample 20.

[0120] exist Figure 3 An exemplary measuring element 30 is shown. The measuring element 30 may be spherical or at least substantially spherical. However, the measuring element 30 is not limited to this. Other shapes or forms may be adopted. In some embodiments, the measuring element 30 may have a diameter of 2 μm to 100 μm. In some embodiments, the measuring element 30 is magnetic. It may be specified that the measuring element 30 is made of, composed of, or contains at least one or more of metals, alloys, plastics, and glass. It may be specified that the core of the measuring element 30 is made of, composed of, or contains the first material, and the surface of the measuring element 30 is made of, composed of, or contains the second material. The first material may be different from the second material. In some embodiments, the measuring element 30 is coated and / or plated.

[0121] The assay element 30 may have one or more capture binding elements 35, to which the analyte 10 may bind or be bound. It may be specified that at least one assay element 30, or several, more, or all assay elements 30, have different types and / or kinds of capture binding elements 35, 36, 37. For example, the assay element 30 may have a first type of capture binding element 35, a second type of capture binding element 36, and / or a third type of capture binding element 37. The number of types and / or kinds of capture binding elements 35, 36, 37 need not be limited to three. In some embodiments, more or fewer different types and / or kinds of capture binding elements 35, 36, 37 may be present. In some embodiments, it may be specified that at least one capture binding element and / or several or all capture binding elements have only one and / or a single type and / or kind of capture binding element 35. In some embodiments, the capture binding elements 35, 36, 37 are tetraspan proteins or include tetraspan proteins, such as CD81, CD63, and / or CD9.

[0122] In some implementations, the number of capture binder elements 35, 36, 37 varies for different measuring elements 30.

[0123] It can be specified that the number of capturing binder elements 35, 36, 37 for each type and / or species is greater than one, see, for example, Figure 3 The capturing binder elements 35, 36, and 37 can extend from the surface of the measuring element 30. The capturing binder elements 35, 36, and 37 can be arranged symmetrically, isotropically, and / or substantially over the entire surface area of ​​the measuring element 30 and / or distributed on the measuring element 30 and / or its surface.

[0124] The capture binder elements 35, 36, and 37 can be selected relative to the analyte 10 to be detected and / or analyzed. In some embodiments, the method and / or apparatus 100 can be configured for multiple determinations and / or heterogeneous determinations when the measuring element 30 has two or more types and / or kinds of capture binder elements 35, 36, and 37, as described below.

[0125] The capture binding elements 35, 36, and 37 may be antibodies for binding analyte 10 or may include antibodies for binding analyte 10. An analyte-bound assay element 30 is formed when analyte 10 binds to at least one of the capture binding elements 35, 36, and 37 of the assay element 30. The analyte 10 may be a ligand and / or antigen and / or a corresponding antibody that binds to the capture binding elements 35, 36, and 37, or may include a ligand and / or antigen and / or a corresponding antibody that binds to the capture binding elements 35, 36, and 37. The capture binding elements 35, 36, and 37 may be selected such that they can bind to a specific, selected, to-be-identified, or desired analyte 10. In particular, when the sample 20 contains more than one type and / or class of analyte 10, the capture binding elements 35, 36, and 37 may be selected, for example, to bind to one of the analytes 10 at a determinable or testable concentration, and / or to one of the analytes 10 that should be measured. It can be specified that when sample 20 includes more than one type and / or class of analyte 10, different types and / or classes of capture binding elements 35, 36, 37 can be selected, for example, selectively bound to one of the analytes 10 at a determinable or testable concentration, and / or selectively bound to one of the analytes 10 that should be measured. In some embodiments, as described below, multiple determinations of multiple analytes 10 (e.g., multiple classes and / or types of analytes 10) are provided.

[0126] exist Figure 4The diagram illustrates an exemplary embodiment of a measuring element 30 according to the present invention. The measuring element 30 has a component test analyte 38. The component test analyte 38 may be disposed on the outer surface of the measuring element 30. The component test analyte 38 may be configured and / or selected such that it can interact with a surface binder element 44 of the substrate 42. In particular, the component test analyte 38 may be bonded to the surface binder element 44. When the component test analyte 38 of the measuring element 30 is bonded to one or more surface binder elements 44 (or vice versa), the speed of the corresponding measuring element 30 changes, particularly decreases.

[0127] Since analyte 10 in sample 20 can also bind to surface binder element 44 of substrate 42, there is competition between the binding interaction between analyte 10 and surface binder element 44 and the binding interaction between element test analyte 38 and surface binder element 44. Specifically, it can be specified that when analyte 10 binds to surface binder element 44, the binding of element test analyte 38 to said surface binder element 44 is reduced or inhibited, and vice versa. In some embodiments, the measuring element 30 can be immobilized (or vice versa) by the interaction between element test analyte 38 and one or more surface binder elements 44.

[0128] Therefore, the higher the concentration of analyte 10 in sample 20, the more analyte 10 binds to the surface binder element 44 of substrate 42, and the less chance or occurrence of element test analyte 38 binding to the surface binder element 44. Thus, the higher the concentration of analyte 10 in sample 20, the less the rate of change and / or decrease of the measuring element 30 with element test analyte 38.

[0129] Competitive measurements can be performed by providing a measuring element 30 with a component test analyte 38.

[0130] exist Figure 5 The image shows another exemplary embodiment of the measuring element 30 according to the present invention. The measuring element 30 has a trapping binder element 35. The trapping binder element 35 may be disposed on the outer surface of the measuring element 30. The trapping binder element 35 may be configured such that the analyte 10 can bind to it. The substrate 42 has a substrate test analyte 47. The substrate test analyte 47 may be disposed on the surface of the substrate 42. The substrate test analyte 47 may be disposed on the substrate 42 such that it can contact the sample 20. The substrate test analyte 47 may be disposed on the substrate 42 such that it can extend into the channel 40.

[0131] The substrate test analyte 47 can be configured and / or selected such that it can interact with the trapping binder element 35 of the measuring element 30. Specifically, the substrate test analyte 47 and the trapping binder element 35 can bind to each other. When the substrate test analyte 47 of the substrate 42 binds to the binder element 35 (or vice versa), the velocity of the corresponding measuring element 30 changes, particularly decreases. In some embodiments, the measuring element 30 can be immobilized (or vice versa) by the interaction between the surface test analyte 47 and one or more surface trapping binder elements 35.

[0132] Since analyte 10 in sample 20 can also bind to the capture binder element 35 of measuring element 30, there is competition between the binding interaction between the analyte and the capture binder element 35 and the binding interaction between the surface test analyte 47 of substrate 42 and the capture binder element 35. Specifically, it can be specified that when analyte 10 binds to the capture binder element 35, the binding of the surface test analyte 47 to the capture binder element 35 is reduced or inhibited, and vice versa.

[0133] Therefore, the higher the concentration of analyte 10 in sample 20, the more analyte 10 binds to the capture binder element 35 of the measuring element 30, and the fewer opportunities or events occur for the capture binder element 35 to bind to the surface test analyte 47. Consequently, the higher the concentration of analyte 10 in sample 20, the less the rate of change and / or decrease of the measuring element 30 with the element test analyte 38.

[0134] Competitive determinations can be performed by providing a substrate 42 with surface test analytes 47.

[0135] This disclosure and the present invention are not necessarily limited to such cases, wherein the substrate 42 and all measuring elements 30 are configured, for example, as shown in the following manner. Figure 3 , Figure 4 and Figure 5 As shown and / or as discussed with reference to these figures. Specifically, it may be specified that in some embodiments, at least one, multiple, or all of the measuring elements 30 have a capture binder element 35 and a component test analyte 38 (not shown in the figures). Alternatively or additionally, some measuring elements 30 may have only the capture binder element 35, and some other measuring elements 30 may have only the component test analyte 38. Alternatively or additionally, the substrate 42 may simultaneously have a surface binder element 44 and a surface test analyte 47 (not shown in the figures). Alternatively or additionally, a substrate 42 may have only the surface binder element 44, and some other substrates 42 may have only the surface test analyte 47. For example, refer to... Figures 1 to 3Any of the embodiments discussed and shown may include component test analyzer 38 and / or surface test analyzer 47, but need not be.

[0136] exist Figure 6 The steps of an exemplary embodiment of the method 1000 according to the present invention are shown. The method may include one or more steps of the method disclosed herein.

[0137] In step 1010, a device 100 having a channel 40 is provided. The channel 40 has a substrate 42. The substrate 42 has surface binder elements 44, 45, 46 configured to bind with analyte 10 in the sample 20, and / or the substrate 42 has a surface test analyte 47. The device 100 may be any device 100 disclosed herein or may include any device 100 disclosed herein.

[0138] In step 1020, the measuring element 30 is mixed into the sample 20. The measuring element 30 can be any measuring element 30 disclosed herein or can include any measuring element 30 disclosed herein. The sample 20 contains analyte 10. The measuring element 30 has capture binder elements 35, 36, 37 configured to bind with analyte 10, and / or the measuring element 30 has element test analyte 38 configured to bind with surface binder elements 44, 45, 46. When the substrate 42 includes surface test analyte 47, the surface test analyte 47 is configured to interact with, and particularly bind to, the capture binder elements 35, 36, 37.

[0139] When the assay element 30 has capture binding elements 35, 36, and 37, the analyte 10 in the sample 20 can bind to the capture binding elements 35, 36, and 37 when or after the assay element 30 is mixed into the sample 20. The capture binding elements 35, 36, and 37 can be one or more antibodies, or may include one or more antibodies. Antibodies can bind to the analyte 10, and / or antibodies can bind to a ligand of the analyte 10, such as an antigen. The capture binding elements 35, 36, and 37 can be selected such that they can bind to a specific, selected, or desired analyte 10.

[0140] When the measuring element 30 has an element test analyte 38, the element test analyte 38 may have properties similar to those of analyte 10 in sample 20. The element test analyte 38 may be of the same type and / or kind as analyte 10 in sample 20. When the substrate 42 has a surface test analyte 47, the surface test analyte 47 may have properties similar to those of analyte 10 in sample 20. The surface test analyte 38 may be of the same type and / or kind as analyte 10 in sample 20.

[0141] In step 1030, the sample 20, which contains the measuring element 30, flows through channel 40. For example, a pump, syringe device, or the like can be used to make the sample 20 flow through channel 40.

[0142] In step 1040, the measuring element 30 moves to the substrate 42. In some embodiments, the measuring element 30 is magnetic, and the measuring element 30 is moved to the substrate 42 by a magnetic field 60. The magnetic field 60 can apply a magnetic force to the measuring element 30, causing the measuring element 30 to move toward the substrate 42. The magnetic field 60 can be a static magnetic field. The measuring element 30 can be pulled or pushed toward the substrate 42.

[0143] In step 1050, the measuring element 30 moves along / on the substrate 42. In some embodiments, the measuring element 30 moves due to the flow of the sample 20. Moving the measuring element 30 along / on the substrate 42 may include rolling the measuring element 30 along / on the substrate 42. In some embodiments, the measuring element 30 may be moved along / on the substrate 42 by a magnetic field 60 or a different magnetic field. The measuring element 30 may be moved along / on the substrate 42 by an external force (such as, for example, a magnetic force). Optionally or additionally, a magnetic field and / or an external force may support the movement of the measuring element 30 along / on the substrate 42.

[0144] As the at least one measuring element 30 moves along / on the substrate 42, the velocity of the at least one measuring element 30 is altered by interaction. This interaction may include, or may be, an interaction, preferably a binding interaction, between at least one surface binder element 44, 45, 46 of the substrate 42, the analyte 10, and at least one trapping binder element 35, 36, 37 of the at least one measuring element 30. Alternatively or additionally, the interaction may include, or may be, an interaction, preferably a binding interaction, between the surface test analyte 47 of the substrate 42 and at least one of the trapping binder elements 35, 36, 37 of the at least one measuring element 30. Alternatively or additionally, the interaction may include, or may be, an interaction, preferably a binding interaction, between the element test analyte 38 of the at least one measuring element 30 and at least one of the surface binder elements 44, 45, 46 of the substrate 42. The velocity change may depend on the strength of the respective interaction.

[0145] Interactions can be such as Figures 1 to 5 Any of the shown and / or references Figures 1 to 5 Any of the interactions described herein may include, for example, Figures 1 to 5 Any of the shown and / or references Figures 1 to 5 Any of the interactions described herein.

[0146] When the speed changes, it can be specified that the measuring element 30 can be fixed or glued to the substrate 42.

[0147] In step 1060, the concentration of analyte 10 is determined by determining the rate change of the measuring element 30 and / or by determining the wear of the measuring element 30. Optionally or additionally, binding kinetics are determined by determining the rate change of the measuring element 30 and / or by determining the wear of the measuring element 30.

[0148] exist Figure 7 The steps of an exemplary embodiment of the method 1000 according to the present invention are shown. The method according to the present invention may be referred to as a rolling bead sandwich determination. The method may include... Figure 7 Additional steps not shown. The method may include one or more steps disclosed herein, and in particular those referenced above. Figure 6 One or more steps of the method described.

[0149] In step 1200, the measuring element 30 is incorporated into the sample 20 containing the analyte 10. Incorporation may include introducing the measuring element 30 into the sample 20. The measuring element 30 may be beads and / or magnetic beads, or may include beads and / or magnetic beads.

[0150] In step 1025, at least one of the assay elements 30 binds to analyte 10 in sample 20. For binding to analyte 10, the assay element 30 has one or more capture binding elements 35, 36, 37. The capture binding elements 35, 36, 37 are configured to bind to analyte 10. Analyte 10 can bind to the capture binding elements 35, 36, 37. Capture binding elements 35, 36, 37 can each be one or more antibodies or may include one or more antibodies. Antibodies can bind to analyte 10, and / or antibodies can bind to a ligand of analyte 10, such as an antigen. Capture binding elements 35, 36, 37 can be selected such that they can bind to a specific, selected, or desired analyte 10.

[0151] When analyte 10 binds to assay element 30, an analyte-bound assay element 30 can be formed, and / or an assay element 30 to which analyte 10 is bound can be designated as an analyte-bound assay element 30. Not all assay elements 30 may or must have analyte 10 bound to capture binder elements 35, 36, 37. Assay elements 30 not bound to analyte 10 may be present in sample 20. However, in some embodiments, all assay elements 30 are analyte-bound assay elements 30 and / or have analyte 10 bound to capture binder elements 35, 36, 37.

[0152] In step 1030, sample 20 flows through channel 40. Sample 20 flowing through channel 40 contains or includes analyte 10 and at least one measuring element 30 bound to analyte 10.

[0153] Channel 40 has a substrate 42. Substrate 42 includes a surface binder element 44. The surface binder element 44 is disposed on the surface of substrate 42, and sample 20 flows along / on the surface of substrate 42, and / or measuring element 30 can move or roll along / on the surface of substrate 42. Surface binder element 44 can bind to analyte 10, and / or analyte 10 can bind to surface binder element 44.

[0154] In step 1040, an external force 60 (e.g., a magnetic field) moves the measuring element 30 in the sample 20 flowing through the channel 40 toward the substrate 42 of the channel 40. The external force 60 may act on the measuring element 30 and may move the measuring element 30 toward or toward the substrate 42 of the channel 40. In some embodiments, the magnetic field 60 applies a (magnetic) force to the measuring element 30.

[0155] It may be specified that moving the measuring element 30 toward or to the substrate 42 includes, is, or corresponds to pulling the measuring element 30 toward or attracting it to the substrate 42. Alternatively or additionally, it may be specified that moving the measuring element 30 toward or to the substrate 42 includes, is, or corresponds to pushing the measuring element 30 to the substrate 42.

[0156] In step 1050, when the measuring element 30 moves to the substrate 42, the measuring element 30 moves or rolls along or on the substrate 42. It may be specified that the measuring element 30 is moved to the substrate 42, for example, to contact the substrate 42 and / or to move or roll along or on the substrate 42, by an external force 60, such as a magnetic force generated by a magnetic field. The measuring element 30 may be pushed / pushed onto the substrate 42 by the flow 110 of the sample 20, such that it can move or roll along or on the substrate 42. In some embodiments, all measuring elements 30 entering the channel 40 are moved by the external force 60, for example, pulled to the substrate 42, such that they move or roll along or on the substrate 42 as they flow through the channel 40. In some embodiments, most or at least a portion of the measuring elements 30 entering the channel 40 are moved by the external force 60, for example, pulled to the substrate 42, such that they move or roll along or on the substrate 42 as they flow through the channel 40.

[0157] The speed of the measuring element 30, bound to the analyte 10, can be varied as it moves or rolls along or on the substrate 42. When the speed is varied, it can decrease. This decrease in speed may be due to interactions, such as binding interactions, between the analyte 10 bound to the measuring element 30 and one or more surface binder elements 44. These binding interactions can be so-called specific binding.

[0158] In some embodiments, altering and / or reducing the speed includes fixing at least one corresponding analyte-bound measuring element 30. In some embodiments, at least one analyte-bound measuring element 30 is slowed down but not fixed. It may be specified that when moving or rolling along or on the substrate 42, measuring elements 30 without analyte 10 bound thereto do not slow down substantially (due to the absence of any binding interactions described in the preceding paragraphs).

[0159] The number and / or proportion of measuring elements 30 with which the corresponding analytes are bound and / or slowed down can depend, for example, the strength of the binding interaction between the surface binder element 44 and the analyte 10 bound to the measuring element 30, and / or the density and / or location of the surface binder elements 44 on the substrate 42, and / or the concentration of the analyte 10 in the sample 20. For example, more than one capturing binder element 35, 36, 37 of the measuring element 30 has analyte 10 bound to it, which in some cases can further reduce the velocity of the measuring element 30 along / on the substrate 42, because multiple binding interactions may exist between the corresponding bound analyte 10 and the multiple surface binder elements 44.

[0160] In step 1060, for example, the concentration of analyte 10 in sample 20 is determined by determining the change and / or decrease in the flow rate of measuring element 30 within channel 40. A higher analyte concentration results in a greater change and / or decrease in the flow rate because more analyte 10 binds to measuring element 30, and measuring element 30 moving or rolling along / on the substrate can be slowed down due to the interaction (e.g., binding interaction) between surface binder element 44 and the analyte 10 bound to measuring element 30. Alternatively or additionally, the concentration of analyte 10 in sample 20 can be determined, for example, by determining the loss of measuring element 30 along channel 40. The loss of measuring element 30 may include the fixation of at least some of the measuring elements 30.

[0161] In this embodiment, sample 20 is flowed or driven by a syringe, syringe pump, and / or metering pump (not shown). The syringe, syringe pump, and / or metering pump may be fluidly connected to channel 40, for example, connected to or connected to the inlet of channel 40. The inlet may be arranged upstream of substrate 42. The syringe, syringe pump, and / or metering pump may apply force to sample 20 such that the velocity of sample 20 is substantially constant as it flows into or through the channel. Therefore, the velocity of measuring element 30 upstream of substrate 42 and / or at the inlet of channel 40 may be known. The velocity of measuring element 30 may be determined, detected, and / or measured by sensor unit 70 downstream of substrate 42. For example, the velocity may be determined or measured by sensor unit 70, as detailed below, for example, by sensing elements 72, 74, 76, and / or by optical device 78. The change and / or decrease in the velocity of measuring element 30 may then be determined by comparing the velocity applied by syringe, syringe pump, and / or metering pump, for example, at the inlet, with the velocity determined by sensor unit 70. In some embodiments, there are no sensing elements 72, 74, 78 and / or optical devices 78 upstream of the substrate 42, and / or the sensing elements 72, 74, 78 and / or optical devices 78 upstream of the substrate 42 are redundant.

[0162] Figures 8 to 11 An exemplary embodiment of the apparatus 100 and method according to the present invention is shown. The apparatus 100 includes a channel 40 having a substrate 42, for example, the channel 40 and substrate 42 as described above.

[0163] Device 100 may include an external force generating device 120 (e.g., a magnetic field generating device). Figure 8 and Figure 9 (not shown in the image) The external force generating device 120 is configured to generate an external force 60, for example, by a magnetic field ( Figure 8 and Figure 9 The device 100 generates a magnetic force (not shown in the diagram). Furthermore, the device 100 includes a sensor unit 70. The sensor unit 70 includes two sensing elements 72 and 74. The second sensing element 74 is arranged downstream of the first sensing element 72, for example, downstream in the flow direction (e.g., in the direction of flow). Figure 8 and Figure 9 (Downstream in the x-direction). For example... Figures 8 to 11 The illustrated embodiments may have at least one, several, or all of the features and / or advantages of at least one other embodiment of the present invention.

[0164] Figure 8 The apparatus 100 is shown, in which a sample 20 containing a measuring element 30 flows through a channel 40 at a first time t1. Figure 9The apparatus 100 is shown, in which a sample 20, including a measuring element 30, flows through a channel 40 at a second time t2. The second time t2 may be later than the first time t1. Figure 10 The outputs of sensor unit 70 and / or first sensing element 72 and second sensing element 74 as a function of time are shown. Figure 11 A comparison is shown between the analyte concentrations detected and / or determined by the method according to the invention and those detected and / or determined by methods of the prior art.

[0165] In some embodiments, sensor unit 70 and / or sensing elements 72, 74 are or include means configured for impedance sensing and / or magnetoresistive sensing. For example, measuring element 30 may be configured to detect and / or sense by impedance sensing and / or by magnetoresistive sensing. Alternatively or additionally, sensor unit 70 and / or sensing elements 72, 74 may be or may include a Coulter counter. In some embodiments, the first sensing element 72 and / or the second sensing element 74 are respectively magnetic field sensor and / or magnetic sensor half-bridges or include magnetic field sensor and / or magnetic sensor half-bridges. Therefore, in some embodiments, non-optical determination of analyte concentration is possible. Thus, the method and apparatus according to the invention allow for determinations in which sample 20 does not need to be treated prior to the method. Furthermore, sample 20 may be opaque.

[0166] The sensor unit 70 and / or sensing elements 72, 74 can detect and / or record the passing, along, or through the measuring element 30, such as the magnetic measuring element 30. It can be specified that the sensor unit 70 and / or sensing elements 72, 74 measure and / or determine the dimensions of the measuring element 30.

[0167] The first sensing element 72 can be arranged upstream of the substrate 42 (see, for example, Figure 8 and Figure 9 In some embodiments, the first sensing element 72 and / or the second sensing element 74 are disposed on / above the substrate 42. The second sensing element 74 may be disposed downstream of the first sensing element 72 and / or downstream of the substrate 42 (see, for example, Figure 8 and Figure 9 The definitions of "downstream" and "upstream" can be relative to the flow direction of flow 110.

[0168] In some implementations, for example in Figures 8 to 11 In one embodiment, the concentration of analyte 10 and / or the decrease in the speed of measuring element 30 can be determined by measuring the rolling time of measuring element 30. Measuring element 30 may be magnetic.

[0169] Rolling time can be determined and / or measured between two sensing elements (e.g., between the first sensing element 72 and the second sensing element 74). However, it should be understood that, additionally or alternatively, rolling time can be determined and / or measured between multiple sensing elements 72, 74, 76 and / or between two sensing elements 72, 74 separated by another sensing element 76. Multiple rolling time measurements and / or determinations can be performed, and optionally or additionally, multiple rolling time measurements and / or determinations can be performed at different temperatures. For example, a first rolling time can be determined and / or measured between the first sensing element 72 and the second sensing element 74, and a second rolling time can be determined and / or measured between the third and fourth sensing elements. For example, the first, second, third, and fourth sensing elements can be different sensing elements. However, it can be specified, for example, that the third sensing element corresponds to or is equivalent to the first or second sensing elements 72, 74. Similarly, different arrangements of sensing elements 72, 74, 76 and / or rolling time measurement and / or determination are possible.

[0170] For example, as depicted, for example, in Figure 8 In this context, the first sensing element 72 can detect and / or record the movement of a measuring element 30, such as a magnetic measuring element 30, along and / or passing through the first sensing element 72 (e.g., moving or rolling along or on the substrate 42). The second sensing element 74 can detect and / or record movement of a measuring element 30, such as a magnetic measuring element 30, along and / or passing through the second sensing element 72 (e.g., moving or rolling along or on the substrate 42, see, for example, ...). Figure 9 The measuring element 30. When one or more measuring elements 30 pass along and / or through one of the sensing elements 72, 74, the corresponding signal can be output by the corresponding sensing element during the passage, see, for example, Figure 10 The more measuring elements 30 and / or the closer the grouped or packaged measuring elements 30 are at a given moment, the higher the signal can be. Therefore, at the first time t1, a first signal corresponding to at least one measuring element 30 or a group of measuring elements 30 passing through the first sensing element 72 can be detected (see...). Figure 8 and Figure 10 Then, when the at least one measuring element 30 or the group of measuring elements 30 passes the second sensing element 74 downstream of the first sensing element 72, the second sensing element 74 may output a second signal at a second time t2 (see, for example, Figure 9 and Figure 10The time difference Δt = t2-t1 can be used in conjunction with the known distance between the first sensing element 72 and the second sensing element 74 to determine the speed of the at least one measuring element 30 or the set of measuring elements 30. The rolling time can be or may correspond to the time difference Δt.

[0171] The higher the analyte concentration in sample 20, the larger the time difference Δt, and the greater the rate decrease. Therefore, the analyte concentration can depend on and / or be related to and / or be a function of the rate decrease, and / or the rate decrease can be used to determine the analyte concentration in sample 20.

[0172] exist Figure 11 In this study, the method according to the invention was compared with conventional assays. While conventional assays can detect and / or determine analyte concentrations on a range of two to three orders of magnitude, the method and apparatus according to the invention can detect and / or determine analyte concentrations on a range of three to four orders of magnitude or even more. Therefore, the method and apparatus according to the invention allow for a significant improvement in the detection and / or determination of analyte concentrations over a wide range, and provide assays with a higher detection range. Figure 30a Another example of concentration determined by the velocity change of measuring element 30 is shown in the figure.

[0173] The reference velocity difference can be specified, for example, based on a given flow rate and / or given a flow rate, and / or determined. For the flow 110 that does not contain analyte 10, i.e., for the measuring element 30 not bound to analyte 10 and / or for the measuring element 30 that does not slow down when moving or rolling along or on the substrate 42, a reference velocity can be determined and / or given. The velocity difference can be determined and / or given relative to the reference velocity. In some embodiments, the reference velocity may be the average flow rate of the flow 110.

[0174] Alternatively or additionally, optical methods may be used to determine the concentration of analyte 10 and / or the velocity of measuring element 30. Sensor unit 70 may be one or more optical devices 78 or may include one or more optical devices 78. Figure 8 and Figure 9 (Not shown in the image), for example, one or more cameras. The sensor unit 70, optical device 78, and / or camera can take snapshots at specific times, and / or capture photographs or videos. The sensor unit 70, optical device 78, and / or camera can capture the movement and / or position of the measuring element 30 in time and / or space. The resolution of the sensor unit 70, optical device 78, and / or camera can be temporally resolved and / or spatially resolved. The sensor unit 70, optical device 78, and / or camera can take snapshots at specific times, and / or capture photographs or videos within the field of view. Field of view (in...) Figure 8and Figure 9 (Not shown) may completely surround the substrate 42 or at least surround a portion of the substrate 42. The field of view may be along the flow direction, for example in... Figure 8 and Figure 9 The x-direction.

[0175] The speed of the measuring element 30 can then be determined, for example, by comparing different positions of the measuring element 30 at different times and optionally or additionally at different temperatures.

[0176] In some embodiments, more than one optical device 78 and / or more than one camera are provided, for example, to have a larger total field of view 79 at sufficient resolution. It may be specified that different optical devices 78 and / or cameras each have different fields of view 79. In some embodiments, at least two, more, or all of the different fields of view 79 at least partially overlap. Alternatively or additionally, it may be specified that at least two, more, or all of the fields of view 79 do not overlap. Different and / or overlapping fields of view 79 can form a composite field of view and / or a total field of view. It may be specified that a first optical device 78 and / or camera is arranged upstream of the substrate 42 and / or such that its field of view 79 covers the upstream region of the substrate 42. A second optical device 78 and / or camera may be arranged downstream of the substrate 42 and / or such that its field of view 79 covers the downstream region of the substrate 42.

[0177] The optical device 78 and / or camera can detect one or more of the color, size, luminescence, and particle size of the measuring element 30. Therefore, in some embodiments, multiple measurements can be achieved.

[0178] exist Figure 12 The diagram illustrates an exemplary signal detected by sensor unit 70 for a single measuring element 30. For example, sensor unit 70 includes a first sensing element 72 and a second sensing element 74. The first sensing element 72 is disposed upstream of the second sensing element 74. When measuring element 30 passes the first sensing element 72 at a first time point, sensing element 70 records a signal, such as... Figure 12 The left spike is shown in the diagram. Then, the measuring element 30 is conveyed downstream, for example by moving along / on the substrate 42 and / or by moving along / on the channel 40, until it passes the second sensing element 74 at a second time point. As the measuring element 30 passes the second sensing element 74, the second sensing element 74 records a signal, such as... Figure 12 The right peak is shown in the image.

[0179] Alternatively or additionally, the corresponding signals can be recorded optically. For example, each spike may correspond to a point in time when the measuring element 30 is at a specific location along the channel 40 and / or the substrate 42, as may be identified, for example, by an optical device such as a camera.

[0180] The first and second time points can be determined and / or measured relative to the same reference time point. In some embodiments, the reference time point can be a time point prior to the first time point. In some embodiments, the reference time point can be the first time point.

[0181] The time difference between the second time point and the first time point can be determined to determine the speed of the measuring element 30, and / or the speed change of the measuring element 30.

[0182] When more than one measuring element 30 is included in the sample 20, and / or when more than one measuring element 30 flows through or moves through the channel 40, a distribution function can be constructed from the records of the sensor unit 70 (e.g., from the records of the first sensor unit 72 and the second sensor unit 74).

[0183] Figure 13 The distribution function constructed from the records of sensor unit 70 is shown. Specifically, the left distribution function 80 ( Figure 13 The left bell-shaped curve (in the image) is constructed from the record of the first sensing element 72. The two distribution functions 82 and 83 on the right (in the image) Figure 13 The solid and dashed bell curves on the right are constructed from the recordings of the second sensing element 72. Alternatively or additionally, distribution functions 80, 82, 83 can be optically determined, for example, by counting the number of measuring elements 30 in a small region at specific time points or within small time intervals. Each distribution function may correspond to a specific location in or along the channel 40 and / or substrate 42. Each distribution function 80, 82, 83 can be determined at a specific location in or along the channel 40 and / or substrate 42.

[0184] like Figure 13 The distribution functions 80, 82, and 83 shown can be or may correspond to histograms and / or density functions. The density function can be a standardized histogram. When the histogram is standardized such that the sum or integral of the histogram over time equals 1, the density function can be interpreted as a probability density function. The density function can be standardized such that, over a time interval between two time points, the integral or sum of the density function over the two time points produces a fraction of the measuring element 30 at a specific location (e.g., at the first or second sensing element 72, 74).

[0185] exist Figure 13 In the diagram, the solid distribution function 82 on the right and the dashed distribution function 83 on the right correspond to different concentrations of analyte 10 in sample 20. On the other hand, in... Figure 13In the example, the left distribution function 80 is the same for the two different concentrations of analyte 10 in sample 20. The left distribution function 80 can be a first distribution function or can correspond to a first distribution function, and either of the right distribution functions 82 and 83 can be a second distribution function or can correspond to a second distribution function.

[0186] The rolling time and / or speed of the measuring element 30 can be determined by the time difference Δt between the peaks of the corresponding distribution function. For example, for a sample 20 with a first concentration of analyte 10, this can be determined by calculation. Figure 13 The time difference is determined by the time difference between the peak value of the solid distribution function 82 on the right and the peak value of the distribution function 80 on the left. For example, for sample 20 with analyte 10 at a second concentration, the time difference can be determined by calculation. Figure 13 The time difference is determined by the time difference between the peak of the distribution function 83 (dashed line on the right) and the peak of the distribution function 80 (dashed line on the left). A peak can be any of the following: (a peak can be any peak that corresponds to a specific maximum value).

[0187] The corresponding difference Δt can indicate the change in rate. The difference between two time differences can indicate the difference in analyte concentration. Similarly, the time difference Δt can be compared with the reference time difference for the reference concentration of the analyte.

[0188] Figure 14 The distribution function constructed from the records of sensor unit 70 is shown. Specifically, the left distribution function 80 ( Figure 14 The left S-shaped curve (in the image) is constructed from the record of the first sensing element 72. The two distribution functions 82 and 83 on the right (in the image) Figure 14 The solid and dashed S-shaped curves on the right are constructed from the recordings of the second sensing element 72. Alternatively or additionally, distribution functions 80, 82, 83 can be optically determined, for example, by counting the number of measuring elements 30 in a small region at specific time points or within small time intervals. Each distribution function may correspond to a specific location in or along the channel 40 and / or substrate 42. Each distribution function 80, 82, 83 can be determined at a specific location in or along the channel 40 and / or substrate 42.

[0189] like Figure 14The distribution function shown can be, or may correspond to, a cumulative function. The distribution function can be, or may correspond to, the empirical cumulative distribution function (ECDF). The cumulative function can be the sum of the values ​​of a histogram or a density function. When the cumulative function is standardized such that its rightmost value equals 1, the cumulative function can be interpreted as a cumulative probability distribution function. The cumulative function can be standardized such that, over a time interval between two time points, the difference in the cumulative function between the two time points produces a fraction of the measuring element 30 at a specific location (e.g., at the first or second sensing element 72, 74).

[0190] exist Figure 14 In the diagram, the solid distribution function 82 on the right and the dashed distribution function 83 on the right correspond to different concentrations of analyte 10 in sample 20. On the other hand, in... Figure 14 In the example, the left distribution function 80 is the same for the two different concentrations of analyte 10 in sample 20. The left distribution function 80 can be a first distribution function or can correspond to a first distribution function, and either of the right distribution functions 82 and 83 can be a second distribution function or can correspond to a second distribution function.

[0191] The rolling time and / or the speed of the measuring element 30 can be determined by the time difference Δt between the inflection points of the corresponding distribution function. For example, for a sample 20 with a first concentration of analyte 10, this can be determined by calculation. Figure 14 The time difference is determined by the time difference between the inflection point of the solid distribution function 82 on the right and the inflection point of the distribution function 80 on the left. For example, for sample 20 with analyte 10 at a second concentration, the time difference can be determined by calculation. Figure 14 The time difference is determined by the difference between the inflection point of the distribution function 83 (dash line on the right) and the inflection point of the distribution function 80 (dash line on the left).

[0192] The corresponding difference Δt can indicate the change in rate. The difference between two time differences Δt can indicate the difference in analyte concentration. Similarly, the time difference Δt can be compared with the reference time difference for the reference concentration of the analyte.

[0193] Figure 15 The distribution function constructed from the records of sensor unit 70 is shown. Specifically, the left distribution function 80 ( Figure 15 The left bell-shaped curve (in the image) is constructed from the record of the first sensing element 72. The two distribution functions 82 and 83 on the right (in the image) Figure 15The solid and dashed bell curves on the right are constructed from the recordings of the second sensing element 72. Alternatively or additionally, distribution functions 80, 82, 83 can be optically determined, for example, by counting the number of measuring elements 30 in a small region at specific time points or within small time intervals. Each distribution function may correspond to a specific location in or along the channel 40 and / or substrate 42. Each distribution function 80, 82, 83 can be determined at a specific location in or along the channel 40 and / or substrate 42.

[0194] like Figure 15 The distribution function shown can be, or may correspond to, a histogram, and / or a density function. The density function can be a standardized histogram. When the histogram is standardized such that the sum or integral of the histogram over time equals 1, the density function can be interpreted as a probability density function. The density function can be standardized such that, over a time interval between two time points, the integral or sum of the density function over the two time points produces a fraction of the measuring element 30 at a specific location (e.g., at the first or second sensing element 72, 74).

[0195] exist Figure 15 In the diagram, the solid distribution function 82 on the right and the dashed distribution function 83 on the right correspond to different concentrations of analyte 10 in sample 20. On the other hand, in... Figure 15 In the example, the left distribution function 80 is the same for the two different concentrations of analyte 10 in sample 20. The left distribution function 80 can be a first distribution function or can correspond to a first distribution function, and either of the right distribution functions 82 and 83 can be a second distribution function or can correspond to a second distribution function.

[0196] The velocity change of the measuring element 30 can be determined by establishing a distance metric between corresponding distribution functions at a specific location. For example, the distance metric can be calculated between the solid line distribution function 82 on the right and the distribution function 80 on the left. This distance metric can then be compared with a reference value. The calculated distance metric can indicate the rolling time and / or velocity change of the sample 20 with a first concentration of analyte 10.

[0197] Similarly, a distance metric can be calculated between the dashed distribution function 83 on the right and the distribution function 80 on the left. This distance metric can then be compared with a reference value. The calculated distance metric can indicate the rolling time and / or speed variation of sample 20 with a second concentration of analyte 20. Figure 15In a specific example, the difference metric for the first concentration can be greater than that for the second concentration because the difference between the solid distribution function on the right and the distribution function on the left is greater than the difference between the dashed distribution function on the right and the distribution function on the left.

[0198] Suitable distance metrics include, but are not limited to, f-divergence, Kullback-Leibler divergence, Wasserstein metric, Kolmogorov-Smirnov test, etc.

[0199] Alternatively or additionally, distribution functions can be compared by fitting a distribution function to a candidate distribution function and comparing the fitted parameters. For example, a candidate distribution function could be a gamma distribution with shape and scale parameters. Similarly, a reference function can be fitted to a candidate distribution function. By comparing the parameters determined from the fitting of each distribution constructed from measurements from the sensor unit, the rolling time and / or the concentration of analyte 10 can be determined.

[0200] For example, Figure 15 The distribution function on the left and Figure 15 The solid distribution functions on the right can all be fitted to candidate distribution functions (as determined for the first analyte concentration). Then, the differences between one or more parameters of the candidate functions can be determined, for example, the differences between the shape parameters of the corresponding gamma functions. Similarly, Figure 15 The distribution function on the left and Figure 15 The dashed distribution function on the right (as determined for the second analyte concentration) can be fitted to a candidate distribution function, and the difference between one or more parameters of the candidate function can be determined, such as the difference between the shape parameters of the corresponding gamma function. The corresponding difference indicates the analyte concentration.

[0201] The candidate distribution function is not limited to the gamma distribution. Other suitable distribution functions can be chosen, such as, but not limited to, the log-normal distribution or the Weibull distribution.

[0202] exist Figure 16 The diagram shows an exemplary curve of the difference in parameters of a candidate distribution function as a function of the analyte concentration in sample 20. In this particular example, the candidate distribution function is a gamma distribution, and the difference corresponds to the difference in shape parameters of the corresponding gamma distributions determined at two different locations in / along channel 40 and / or substrate 42.

[0203] In this specific example, the analyte concentration increases as the parameter difference decreases (in this case, as the corresponding shape parameter difference decreases). Therefore, the parameter difference indicates the analyte concentration. The analyte concentration can be determined from the parameter difference.

[0204] Figure 17and Figure 18 Another embodiment of the apparatus 100 and method according to the present invention is shown. Figure 17 The apparatus 100 is shown, in which a sample 20 containing a measuring element 30 flows through a channel 40. Figure 18 A comparison is shown between the analyte concentration detected and / or determined by the method according to the invention and the analyte concentration detected and / or determined by a prior art method. The measuring element 30 may be magnetic, and / or may include a magnetic measuring element, such as a magnetic bead.

[0205] Device 100 includes a channel 40 having a base 42, for example, the channel 40 and base 42 as described above. Device 100 may have an external force generating device 120, such as a magnetic field generating device. Figure 17 (not shown), which is configured to generate an external force 60, such as by a magnetic field ( Figure 17 The device 100 generates a magnetic force (not shown in the diagram). Furthermore, the device 100 includes a sensor unit 70. The sensor unit 70 includes two sensing elements 72 and 74. The second sensing element 74 can be arranged downstream of the first sensing element 72, for example, downstream in the flow direction (e.g., in the direction shown in the diagram). Figure 17 Downstream in the x-direction). For example... Figure 17 and Figure 18 The illustrated embodiments may have one, several, or all of the features and / or advantages of at least one other embodiment of the present invention.

[0206] The concentration of analyte 10 in sample 20 can be determined by measuring the loss of the measuring element 30 along channel 40 (e.g., the loss of a magnetic measuring element), see, for example, as Figure 17 and Figure 18 The implementation scheme shown is illustrated.

[0207] For example, loss determination can be performed using differential counting. For example, sensor unit 70, first sensing element 72, and / or second sensing element 74 can be specified to detect, record, and / or count passing through, along, or across measuring element 30, such as a magnetic measuring element. In some embodiments, sensor unit 70 and / or sensing elements 72, 74 are or include means configured for impedance sensing and / or magnetoresistive sensing. Measuring element 30 can be configured to detect and / or sense by impedance sensing and / or by magnetoresistive sensing. Optionally or additionally, sensor unit 70 and / or sensing elements 72, 74 can be or include a Coulter counter. In some embodiments, first sensing element 72 and / or second sensing element 74 are respectively magnetic field sensor and / or magnetic sensor half-bridges or include magnetic field sensor and / or magnetic sensor half-bridges.

[0208] The differential count between two sensing elements can be determined and / or measured, for example, the differential count between the first sensing element 72 and the second sensing element 74, see, for example, Figure 17 Differential counting can be based on a comparison of the counts of measuring element 30 by two different and / or differentiated sensing elements 72, 74. It can be specified that the sensing element outputs a signal that can be used for counting and / or corresponding to the counts of measuring element 30. "Counting by sensor unit and / or sensing element" can include the sensor unit and / or sensing element outputting a suitable signal, and the counting being performed by a control unit and / or a computer unit. Optionally or additionally, differential counting can be repeated at different temperatures.

[0209] However, it should be understood that differential counts can be determined and / or measured between multiple sensor elements 72, 74, 76, and / or between two sensor elements 72, 74 separated by another sensor element 76. Multiple differential counts can be performed. For example, a differential count can be performed by comparing the number of measuring elements 30 counted by the first sensing element 72 with the number of measuring elements 30 counted by the second sensing element 74, and a second differential count can be performed by comparing the count of the third sensing element with the count of the fourth sensing element. For example, the first, second, third, and fourth sensing elements can be different sensing elements 72, 74, 76. However, it can be specified, for example, that the third sensing element corresponds to or is equivalent to the first or second sensing element 72, 74. Similarly, different arrangements of sensing elements 72, 74, 76 and / or differential counts are possible. A differential count can be or may correspond to the difference in the number of measuring elements 30 counted by two corresponding sensing elements 72, 74.

[0210] The loss of measuring element 30 may include the fixation of at least some of the measuring elements 30. For example, when measuring elements 30 to which analyte 10 is bonded, for example, to and / or fixed to substrate 42 by one or more surface binder elements 44, sensing elements 72, 74 may count different numbers of measuring elements 30. Since the number and / or proportion of measuring elements 30 fixed and / or bonded to substrate 42 may depend on the analyte concentration, a higher differential count indicates a higher analyte concentration. Therefore, the analyte concentration may depend on the differential count, may be related to the differential count and / or may be a function of the differential count, and / or may be used to determine the analyte concentration.

[0211] exist Figure 18In this study, the method according to the invention is compared with conventional assays. While conventional assays can detect and / or determine analyte concentrations on two to three orders of magnitude, the method and apparatus according to the invention can detect and / or determine analyte concentrations on three to four or even more orders of magnitude. Therefore, the method and apparatus according to the invention allow for significantly improved detection and / or determination of analyte concentrations over a wide range and provide assays with a higher detection range. It can be specified that... Figure 30b , Figure 31a and / or Figure 31b The proportion and / or number of the measuring elements 30 that are stuck and / or fixed are determined by differential counting.

[0212] Alternatively or additionally, optical methods may be used to determine the concentration of the analyte and / or to perform differential counting. Sensor unit 70 may be one or more optical devices 78 or may include one or more optical devices 78. Figure 17 (Not shown in the image), such as one or more cameras. The sensor unit 70, optical device 78, and / or camera can take snapshots at specific times, and / or capture photos or videos. The sensor unit 70, optical device 78, and / or camera can capture the movement and / or position of the measuring element 30 in time and / or space. The sensor unit 70, optical device 78, and / or camera can take snapshots at specific times, and / or capture photos or videos in the field of view 79.

[0213] In some embodiments, more than one optical device 78 and / or more than one camera are provided, for example, to have a larger total field of view 79 at sufficient resolution. It may be specified that the different optical devices 78 and / or cameras each have a different field of view 79. In some embodiments, at least two, more, or all of the different fields of view 79 may at least partially overlap. Alternatively or additionally, it may be specified that at least two, more, or all of the fields of view 79 do not overlap. Field of view ( Figure 17 (Not shown) may completely surround the substrate 42 or surround at least a portion thereof. The field of view may be along the flow direction, for example, Figure 17 The x-direction. Different and / or overlapping fields of view 79 can form a composite field of view and / or a total field of view. It can be specified that a first optical device 78 and / or camera is arranged upstream of the substrate 42 and / or such that its field of view 79 covers the upstream region of the substrate 42. A second optical device 78 and / or camera can be arranged downstream of the substrate 42 and / or such that its field of view 79 covers the downstream region of the substrate 42. The optical device 78 and / or camera can detect one or more of the color, size, luminescence, and particle size of the measuring element 30. Therefore, in some embodiments, multiple measurements are possible.

[0214] For example, the movement and / or position of the measuring element 30 (e.g., a magnetic measuring element) in the channel 40 and / or on the substrate 42 can be captured, for example, movement or rolling along or on the substrate 42. The velocity and / or position of the measuring element 30 can then be determined, for example, by comparing different positions of the measuring element 30 at different times. Fixed and / or stuck measuring elements 30 can be identified and / or determined, for example, by comparing positions and / or velocities at different times. Differential counting can include, may correspond to, and / or may be equivalent to, the number of fixed and / or stuck measuring elements 30.

[0215] Figures 19 to 21 Another embodiment of the apparatus 100 and method according to the present invention is shown. For example, the concentration of analyte 10 and / or the wear of measuring element 30 (e.g., wear of magnetic measuring element) can be determined by detecting the shaking of measuring element 30. Figure 20 The detected shaking is shown, which occurs when the measuring element 30 is glued and / or fixed to, for example, the substrate 42. Figure 21 A comparison is shown between analyte concentrations detected and / or determined by the method according to the invention and analyte concentrations detected and / or determined by methods of the prior art. Figures 19 to 21 The illustrated embodiments may have one, several, or all of the features and / or advantages of at least one other embodiment of the present invention.

[0216] Figure 19 An apparatus 100 is shown in which a sample 20, including a measuring element 30, flows through a channel 40. The apparatus 100 includes a channel 40 having a substrate 42, for example, the channel 40 and / or substrate 42 as described above. The apparatus 100 may include an external force generating device 120 (e.g., a magnetic field generating device). Figure 19 (not shown in the image), the external force generating device 120 is configured to generate an external force 60 ( Figure 19 (Not shown in the image), for example, magnetic force generated by a magnetic field. Furthermore, the device 100 includes a sensor unit 70. The sensor unit 70 includes one or more optical devices 78. The optical devices 78 may be one or more cameras, or may include one or more cameras. The sensor unit 70, optical devices 78, and / or cameras can take snapshots at specific times, and / or capture photographs or videos. The sensor unit 70, optical devices 78, and / or cameras can capture the movement and / or position of the measuring element 30 in time and / or space.

[0217] In some embodiments, more than one optical device 78 and / or more than one camera are provided, for example, to have a larger total field of view 79 at sufficient resolution. It may be specified that the different optical devices 78 and / or cameras each have a different field of view 79. In some embodiments, at least two, more, or all of the different fields of view 79 at least partially overlap. Alternatively or additionally, it may be specified that at least two, more, or all of the fields of view 79 do not overlap. Different and / or overlapping fields of view 79 can form a composite field of view and / or a total field of view.

[0218] The field of view 79 may completely surround the substrate 42 or surround at least a portion thereof. The field of view 79 may be along the flow direction, for example, along... Figure 19 The x-direction. It can be specified that the first optical device 78 and / or camera is arranged upstream of the substrate 42 and / or such that its field of view 79 covers the upstream region of the substrate 42. The second optical device 78 and / or camera can be arranged downstream of the substrate 42 and / or such that its field of view 79 covers the downstream region of the substrate 42. The optical device 78 and / or camera can detect one or more of the color, size, luminescence, and particle size of the measuring element 30. Therefore, in some embodiments, multiple measurements can be achieved.

[0219] In some embodiments, the concentration of analyte 10 is determined by detecting the agitation of measuring element 30 (e.g., agitation of magnetic measuring element 30), which may occur when bound to at least one of the surface binder elements 44, see, for example, Figures 19 to 21 The illustrated implementation scheme. Shaking can be detected by optical means, such as, for example, using... Figure 19 The optical device 78 in the middle.

[0220] When the analyte-bonded measuring element 30 is adhered to and / or fixed to the substrate 42, the bonding interaction can apply a force to the measuring element 30, which moves or rolls along or on the substrate 42, causing the measuring element 30 to vibrate. This vibration can be detected by the sensor unit 70, for example, optically and / or by an optical device 78, see, for example, Figure 19 .exist Figure 20 In the signal spike, the signal spike corresponds to the binding of the analyte binding measuring element 30 to at least one surface binder element 44, and / or corresponds to the shaking of the measuring element 30.

[0221] The shaking of the detection element 30 can correspond to, involve, and / or be related to differential counting. Therefore, the concentration of analyte 10 can be determined by detecting shaking and / or by counting the number of shaking events. The analyte concentration can depend on shaking, can be related to shaking, and / or can be a function of shaking, and / or the analyte concentration can be determined using the detection of shaking.

[0222] exist Figure 21 In this study, the method according to the invention is compared with conventional assays. While conventional analyses can detect and / or determine analyte concentrations over two to three orders of magnitude, the method and apparatus according to the invention can detect and / or determine analyte concentrations over three to four or even more orders of magnitude. Therefore, the method and apparatus according to the invention allow for a significant improvement in the detection and / or determination of analyte concentrations over a wide range and provide assays with a higher detection range. It can be specified that the determination is made by detecting the shaking of the measuring element 30. Figure 30b , Figure 31a and / or Figure 31b The proportion and / or number of the measuring elements 30 that are glued and / or fixed.

[0223] Figure 22 A device 100 according to an embodiment of the present invention is shown, in which a stratified flow 114 is employed. The stratified flow may be a sheath flow or may include a sheath flow. The device 100 includes a channel 40 having a substrate 42, for example, a channel 40 and a substrate 42 as described above. The substrate 42 includes a surface binder element 44. The device 100 may include an external force generating device 120, for example, a magnetic field generating device 120. The external force generating device 120 may be configured to generate an external force 60 and / or a magnetic field.

[0224] Layered flow 114 can convert sample flow 112 (which can correspond to) Figures 1 to 21 The sample flow 112 (114) is separated from the substrate 42 in the channel 40. In some embodiments, the stratified flow 114 separates the sample flow 112 from the substrate 42 in the channel 40 along the entire length of the channel 40 or at least partially. In some embodiments, the stratified flow 114 separates the sample flow 112 from the substrate 42 in the channel 40 at least above / above the substrate 42. In some embodiments, more than one stratified flow 114 is provided. Figure 22 (Not shown in the image). If more than one layered flow 114 is provided, the layered flows can be arranged to flow above and / or over each other. The layered flows can flow substantially in corresponding planes parallel to the surface of the base 42, wherein said planes can be arranged above each other with respect to a direction perpendicular to the surface of the base 42.

[0225] An external force 60 (e.g., a magnetic force generated by a magnetic field) can be configured to move the measuring element 30 (e.g., a magnetic measuring element) from the sample stream 112 into the stratified stream 114, and / or through the stratified stream 114 toward the substrate 42. The external force 60 can pull or attract the measuring element 30 from the sample stream 112 into the stratified stream 114, and / or pull or attract the measuring element 30 toward the substrate 42 through the stratified stream 114. The height of the stratified stream 114 (e.g., at...) Figure 22The diameter of the measuring element 30 (in the z direction) can be greater than the diameter of the measuring element 30, or greater than the diameter of the largest of the measuring elements 30.

[0226] By using the stratified flow 114, interference from background cells or debris in the sample 20 can be prevented from moving or rolling the measuring element 30. The stratified flow 114 can reduce or minimize interference from hematocrit.

[0227] Figures 23 to 25 Embodiments of the apparatus 100 and method according to the invention are shown, configured for and / or for multiplex assays. The method and apparatus according to the invention can be adapted to perform heterogeneous assays. It can be specified that sample 20 comprises or contains more than one and / or multiple different kinds or types of analytes 10, at least one or more of which, or corresponding concentrations, will be detected and / or analyzed. For example, in Figure 23 , Figure 24 and / or Figure 25 The diagram illustrates different exemplary embodiments that can determine the concentration of different types and / or species of analytes 10.

[0228] Figure 23 , Figure 24 and / or Figure 25 Each device 100 includes a corresponding channel 40 having a substrate 42 and a corresponding sensor unit 70. Figures 23 to 25 (Not shown in the image). Device 100 may include a corresponding external force generating device 120 ( Figures 23 to 25 (Not shown in the image), for example, includes a magnetic field generating device to generate a magnetic field. Such as... Figure 23 , Figure 24 and / or Figure 25 The illustrated embodiments may have one, more, or all of the features and / or advantages of at least one other embodiment of the invention. The measuring element 30 may be a magnetic measuring element or may include a magnetic measuring element.

[0229] For different types and / or kinds of analytes 10 to be detected and / or analyzed, the substrate 42 may have different types and / or kinds of surface binder elements 44, 45, 46, see, for example, Figure 23 , Figure 24 and Figure 25For example, substrate 42 may include a first surface binder element 44, a second surface binder element 45, and / or a third surface binder element 46. The first surface binder element 44 may be configured to bind to a first type and / or class of analyte 10, the second surface binder element 45 may be configured to bind to a second type and / or class of analyte 10, and / or the third surface binder element 46 may be configured to bind to a third type and / or class of analyte 10. The different types and / or classes of surface binder elements 44, 45, 46 may be arranged, for example, one after another, alternately, and / or substantially randomly. In some embodiments, the different types and / or classes of surface binder elements 44, 45, 46 are arranged in a homogeneous group on a portion of substrate 42, see, for example, Figure 25 It can be specified that homogeneous groups are separated by sensing element 76, see, for example, Figure 25 The implementation plan.

[0230] The measuring element 30 may have different sizes or diameters, see, for example, Figure 23 It can be specified that all measuring elements 30 having substantially the same size or diameter have the same type and / or kind of capture binder elements 35, 36, 37 and / or can be configured to bind the same type and / or kind of analyte 10. However, measuring elements 30 having different sizes or diameters may have different types and / or kinds of capture binder elements 35, 36, 37 and / or can be configured to bind different types and / or kinds of analytes 10. Measuring elements 30 can be grouped into specific and / or different groups according to their size or diameter. It can be specified that all measuring elements 30 in the same group have the same type and / or kind of capture binder elements 35, 36, 37. It can be specified that all measuring elements 30 in the same group having substantially the same size or diameter are substantially identical or have substantially the same characteristics.

[0231] For example, a measuring element 30 having a first size or diameter may have a first trapping binder element 35, a measuring element 30 having a second size or diameter may have a second trapping binder element 36, and / or a measuring element 30 having a third size or diameter may have a third trapping binder element 37. The first trapping binder element 35, the second trapping binder element 36, and / or the third trapping binder element 37 may be different. Therefore, a measuring element 30 with a first size or diameter may be configured to be combined with a first type and / or species of analyte 10, a measuring element 30 with a second size or diameter may be configured to be combined with a second type and / or species of analyte 10, and a measuring element 30 with a third size or diameter may be configured to be combined with a third type and / or species of analyte 10.

[0232] Alternatively or additionally, the measuring element 30 may have a different magnetic moment, see, for example, Figure 24 For example, some of the measuring elements 30 may be or may contain materials different from those of some of the other measuring elements 30, such that their respective magnetic moments may be different.

[0233] The measuring elements 30 can be grouped into specific and / or different groups based on their magnetic moments. It can be specified that all measuring elements 30 in the same group have the same kind and / or type of trapping binder elements 35, 36, 37. It can also be specified that all measuring elements 30 in the same group have substantially the same magnetic moment, are substantially identical, or have substantially the same characteristics.

[0234] It can be specified that all measuring elements 30 having substantially the same magnetic moment have the same type and / or kind of capture binder elements 35, 36, 37 and / or are configured to bind the same type and / or kind of analyte 10. However, measuring elements 30 with different magnetic moments may have different types and / or kinds of capture binder elements 35, 36, 37 and / or may be configured to bind different types and / or kinds of analytes 10. For example, a measuring element 30 having a first magnetic moment may have a first capture binder element 35, a measuring element 30 having a second magnetic moment may have a second capture binder element 36, and / or a measuring element 30 having a third magnetic moment may have a third capture binder element 37. The first capture binder element 35, the second capture binder element 36, and / or the third capture binder element 37 may be different. Therefore, the first magnetic moment measuring element 30 can be configured to be combined with a first type and / or type of analyte 10, the second magnetic moment measuring element 30 can be configured to be combined with a second type and / or type of analyte 10, and the third magnetic moment measuring element 30 can be configured to be combined with a third type and / or type of analyte 10.

[0235] It can be specified that some or a group of measuring elements 30 have the same size, but different magnetic moments. Alternatively, some or a group of measuring elements 30 may differ in both size and magnetic moment.

[0236] When a measuring element 30 of a specific size and / or a specific magnetic moment has specific trapping binder elements 35, 36, 37, for example, to bind to a specific analyte 10, the concentration of different specific analytes 10 can be determined by utilizing the rate of decrease of measuring elements 30 of different sizes and / or different magnetic moments. Therefore, multiple determinations and / or differentiation between different analytes 10 are possible.

[0237] Measuring elements 30 of different sizes or diameters can be distinguished by sensor unit 70 and / or by sensing elements 72, 74, 76. Similarly, measuring elements 30 of different magnetic moments can be distinguished by sensor unit 70 and / or by sensing elements 72, 74, 76. Optionally and / or additionally, optical devices 78 can be used, for example, to determine the size and / or velocity of the measuring element 30, and thereby determine the associated analyte concentration.

[0238] Optionally or additionally, at least two measuring elements may have different acoustic characteristics and different electrical characteristics. It may be specified that measuring elements can be distinguished or grouped based on their different acoustic characteristics, different electrical characteristics, and / or different optical characteristics.

[0239] Optical properties may include color or markings, such that, for example, the measuring element may be colored or may be a marked measuring element. Acoustic properties may include compressibility and / or density, such as, for example, the compressibility and / or density of the measuring element. Electrical properties may include impedance and / or charge, such as, for example, the impedance and / or charge of the measuring element.

[0240] In some embodiments, measuring elements can be distinguished, grouped, and / or identified based on their acoustic, electrical, and / or optical characteristics. For example, device 100 can be configured to detect acoustic, electrical, and / or optical characteristics. In some embodiments, sensor unit 70 is configured to detect acoustic, electrical, and / or optical characteristics. In some embodiments, optical device 78 can detect measuring elements of different colors or markings.

[0241] When a measuring element 30 with specific acoustic, electrical, and / or optical properties has specific trapping binder elements 35, 36, and 37, for example, bound to a specific analyte 10, the concentration of different specific analytes 10 can be determined by utilizing the rate of decrease of the measuring elements 30 with different acoustic, electrical, and / or optical properties. Therefore, multiple determinations and / or differentiation between different analytes 10 are possible.

[0242] Therefore, see, for example, Figure 23 and Figure 24 The concentrations of the corresponding different types and / or species of analytes 10 can be determined by measuring the decrease in the velocity of the measuring element 30 along the channel 40 and / or by measuring the wear of the measuring element 30, as described above. For this purpose, at least two sensing elements 72, 74 can be provided. Figure 22 , Figure 23 and Figure 24(Not shown in the diagram), for example, a first sensing element 72 is arranged upstream of surface binder elements 44, 45, 46, and a second sensing element 74 is arranged downstream of surface binder elements 44, 45, 46. Since the sensor unit 70 and / or sensing elements 72, 74 can distinguish different types of measuring elements 30 (e.g., defined by their different sizes and / or different magnetic moments), a decrease in the speed of the measuring elements 30 and / or the wear of each measuring element 30 of different sizes and / or different magnetic moments can be detected; for example, each group of measuring elements 30 of different sizes and / or different magnetic moments can be detected. Since measuring elements 30 of different sizes and / or different magnetic moments are associated with different types and / or kinds of analytes 10, the concentration of analytes 10 can be determined as described above. Alternatively or additionally, an optical device 78 can be used, for example, to determine the speed, number, and / or size of the measuring elements 30, and thereby determine the associated analyte concentration.

[0243] Similarly, the rate of decrease in loss of measuring element 30 with different acoustic, electrical and / or optical properties can be determined.

[0244] Alternatively, refer to Figure 25 It can be specified that the measuring element 30, all measuring elements 30, or at least some measuring elements 30 have multiple different types and / or kinds of trapping binder elements 35, 36, 37, or groups of different kinds and / or kinds of trapping binder elements 35, 36, 37. It can also be specified that the measuring element 30, all of the measuring elements 30, or at least some of the measuring elements 30 are neither different in size or diameter nor in magnetic moment (or do not have different acoustic, electrical, and / or optical properties), but have multiple different types and / or kinds of trapping binder elements 35, 36, 37, and / or groups of different kinds and / or kinds of trapping binder elements 35, 36, 37. For example, the measuring element 30 may have one or more first type and / or kinds of trapping binder elements 35, one or more second type and / or kinds of trapping binder elements 36, and / or one or more third type and / or kinds of trapping binder elements 37. The first capture binding element 35, the second capture binding element 36, and / or the third capture binding element 37 may be different. The first capture binding element 35 may be configured to bind to a first type and / or class of analyte 10, the second capture binding element 36 may be configured to bind to a second type and / or class of analyte 10, and the third capture binding element 37 may be configured to bind to a third type and / or class of analyte 10. Accordingly, the measuring element 30 may capture the first type and / or class of analyte 10, the second type and / or class of analyte 10, and / or the third type and / or class of analyte 10.

[0245] exist Figure 25 In this embodiment, multiple determinations and / or differentiation between different analytes 10 are performed by providing a homogeneous group of surface binder elements 44, 45, 46. A specific homogeneous group of surface binder elements 44, 45, 46 can be configured to bind to a specific analyte 10. The concentration of the corresponding specific analyte 10 can then be determined by utilizing the decrease in the velocity of the measuring element 30 and / or the measuring element 30 bound to the analyte along the substrate 42 in the region of the specific homogeneous group / on the substrate 42 in the region of the specific homogeneous group. Therefore, multiple determinations and / or differentiation between different analytes 10 are possible.

[0246] The decrease in velocity of the sensing element 30 along the channel and / or the loss of the sensing element 30 can be determined by two sensing elements 76 separated by the homogeneous surface binder elements 44, 45, 46, for example, as Figure 25 As shown.

[0247] For example, when the first type of analyte 10 can be bonded to the first surface binder element 44, the measuring element 30 bonded to the first analyte 10 can be slowed down and / or fixed to the substrate 42 in the region of the first surface binder element 44 of the homogeneous group. Therefore, to determine the analyte concentration of the first type and / or species of analyte 10, the decrease in the velocity of the measuring element 30 along the channel 40 and / or the loss of the measuring element 30 can be determined as described above by a first sensing element 76 and a second sensing element 76, the first sensing element 76 being arranged upstream and adjacent to the first surface binder element 44 of the homogeneous group, and the second sensing element 76 being arranged downstream and adjacent to the first surface binder element 44 of the homogeneous group. In other words, the change and / or decrease in the velocity of the measuring element 30 along the channel 40, and / or the loss of the measuring element 30, can be determined by the sensing element 76 sandwiching the first surface binder element 44 of the homogeneous group in the middle.

[0248] Similarly, when the second type of analyte 10 can be bound to the second surface binder element 45, the measuring element 30 bound to the second analyte 10 can be slowed down and / or fixed to the substrate 42 in the region of the second surface binder element 45 of the homogeneous group. Therefore, to determine the analyte concentration of the second type and / or class of analyte 10, the reduction in the velocity of the measuring element 30 along the channel 40 and / or the wear of the measuring element 30 can be determined as described above by a first sensing element 76 and a second sensing element 76, the first sensing element 76 being arranged upstream and adjacent to the second surface binder element 45 of the homogeneous group, and the second sensing element 76 being arranged downstream and adjacent to the second surface binder element 45 of the homogeneous group. In other words, the reduction in the velocity of the measuring element 30 along the channel 40 and / or the wear of the measuring element 30 can be determined by the sensing element 76 sandwiching the second surface binder element 45 of the homogeneous group in the middle.

[0249] Similarly, when the third type of analyte 10 can be bound to the third surface binder element 46, the measuring element 30 bound to the third analyte 10 can be slowed down and / or fixed to the substrate 42 in the region of the third surface binder element 46 of the homogeneous group. Therefore, to determine the analyte concentration of the third type and / or species of analyte 10, the reduction in the velocity of the measuring element 30 along the channel 40 and / or the wear of the measuring element 30 can be determined as described above by a first sensing element 76 and a second sensing element 76, the first sensing element 76 being arranged upstream and adjacent to the third surface binder element 46 of the homogeneous group, and the second sensing element 76 being arranged downstream and adjacent to the third surface binder element 46 of the homogeneous group. In other words, the reduction in the velocity of the measuring element 30 along the channel 40 and / or the wear of the measuring element 30 can be determined by sensing elements 76 sandwiching the third surface binder element 46 of the homogeneous group in the middle.

[0250] Alternatively or additionally, one or more optical devices 78 may be used, for example, to determine the size, speed and / or number of measuring elements 30 in the regions of surface binder elements 44, 45, 46 of corresponding homogeneous groups, and thereby determine the associated analyte concentration.

[0251] Although reference Figure 23 , Figure 24 and Figure 25Three different types and / or varieties of analytes 10, measuring elements 30, capture binder elements 35, 36, 37, and / or surface binder elements 44, 45, 46 are described, and the number of analytes 10, measuring elements 30, capture binder elements 35, 36, 37, and / or surface binder elements 44, 45, 46 is not limited thereto. Similarly, the arrangement of surface binder elements 44, 45, 46 can be different and / or not limited thereto. It can be specified that the different measuring elements 30 characterized as described above and / or above are mixed and / or used.

[0252] In some embodiments, at least one, more, or all of the capture binding elements 35, 36, 37 are tetraspan proteins or include tetraspan proteins, such as CD81, CD63, and / or CD9. In some embodiments, at least one, more, or all of the surface binding elements 44, 45, 46 are tetraspan proteins or include tetraspan proteins, such as CD81, CD63, and / or CD9.

[0253] It can be specified that the capturing binder elements 35, 36, and 37 are different, but the surface binder elements 44, 45, and 46 can be the same, i.e., of the same kind and / or type. In some embodiments, the surface binder elements 44, 45, and 46 can be universal for all target analytes 10.

[0254] In some embodiments, it may be specified that there is only one type and / or class of capture binding elements 35, 36, 37, such as an antibody of a certain type or class for all capture binding elements 35, 36, 37, and only one type and / or class of surface binding elements 44, 45, 46, such as an antibody of a certain type or class for all surface binding elements 44, 45, 46. The type and / or class of capture binding elements 35, 36, 37 may be the same as the type and / or class of surface binding elements 44, 45, 46. Alternatively, the type and / or class of capture binding elements 35, 36, 37 may be different from the type and / or class of surface binding elements 44, 45, 46. In some embodiments, it may be specified that at least one, more, or all of the capture binding elements 35, 36, 37 of the assay element 30 are of the same type and / or class as at least some, more, or all of the surface binding elements 44, 45, 46.

[0255] It can be specified that all capture binding elements 35, 36, and 37 of the assay element 30 are identical, i.e., of the same kind and / or type, but the surface binding elements 44, 45, and 46 are different. In some embodiments, at least one, more, or all of the capture binding elements 35, 36, and 37 of the assay element 30 are matched with at least one, more, or all of the surface binding elements 44, 45, and 46, such that the corresponding capture binding elements 35, 36, and 37 and surface binding elements 44, 45, and 46 form matched antibody pairs.

[0256] It can be stipulated, as per reference Figure 23 , Figure 24 and Figure 25 The aforementioned embodiments are combined. For example, surface binder elements 44, 45, and 46 can be arranged in a homogeneous group, and the dimensions and / or magnetic moments (or acoustic, electrical, and / or optical properties) of measuring element 30 can be different.

[0257] When the measuring elements 30 are spherical or substantially spherical, they can move or roll along or on the substrate 42 such that substantially identical portions of their surfaces repeatedly contact the substrate 42 during movement or rolling. For example, the measuring elements 30 can move or roll substantially along a circumferential disk on their surface. Therefore, the analytes 10 bound to the trapping binder elements 35, 36, 37 arranged outside the circumferential disk may not contact the substrate 42 and / or the surface binder elements 44, 45, 46 at all, or at least not as frequently as the analytes 10 bound to the trapping binder elements 35, 36, 37 arranged on or within the circumferential disk.

[0258] Therefore, in some embodiments, it may be specified that the measuring element 30, which moves or rolls on the substrate 42, is manipulated, and / or the orientation or direction of movement or rolling of the measuring element is changed. Figure 26 , Figure 27 and Figure 28 Some corresponding exemplary embodiments according to the present invention are shown.

[0259] like Figure 26 , Figure 27 and Figure 28 The illustrated embodiments include at least one, several, and / or all of the features and / or advantages of the apparatus 100 and / or method as described herein, such as, but not limited to, those described above and / or... Figures 1 to 25 Those shown. In particular, such as Figure 26 , Figure 27 and Figure 28The illustrated embodiment includes or comprises one or more of the following: channel 40, external force generating device 120 (e.g., magnetic field generating device), analyte 10, and / or sample 20, for example, Figure 1 Or as shown in 2, they were not in Figure 26 , Figure 27 and / or Figure 28 As shown in the figure. The measuring element 30 may be magnetic, and / or may include a magnetic measuring element.

[0260] In some implementations, channel 40 has a pattern 48, as can be seen in, for example Figure 26 , Figure 27 and Figure 28 Pattern 48 can be configured to alter, manipulate, or change the movement, direction, and / or orientation of the measuring element 30, for example, when flowing within the channel 40 and / or when moving or rolling along or on the substrate 42. Pattern 48 can alter, manipulate, or change the movement, direction, and / or orientation of the measuring element 30 to be perpendicular to the flow direction or at least at an angle greater than 0 degrees, preferably 10 degrees or less, more preferably 6 degrees or less, for example in… Figure 26 , Figure 27 and Figure 28 In the x-direction. In some embodiments, the angle is a maximum of 15°, more preferably a maximum of 10°. Pattern 48 can deflect and / or turn the measuring element 30 that flows within the channel 40 and / or moves or rolls along / on the substrate 42. The pattern can apply a force to the measuring element 30, resulting in a change in movement, direction, or orientation. Pattern 48 can be a mechanical and / or magnetic structure adapted to manipulate and / or change the movement, direction, and / or orientation of the measuring element 30 relative to the flow direction, or may include a mechanical and / or magnetic structure adapted to manipulate and / or change the movement, direction, and / or orientation of the measuring element 30 relative to the flow direction.

[0261] Pattern 48 can be a mechanical pattern 48. Mechanical pattern 48 can be and / or may include notches, grooves, ridges, etc. Optionally or additionally, pattern 48 can be a magnetic pattern 48. Magnetic pattern 48 can apply a magnetic force to the measuring element 30 (e.g., as shown in the image). Figure 28 (As shown by reference numeral 130 in the accompanying drawings), for example, to manipulate or change the movement, direction, or orientation of the measuring element 30. The magnetic pattern 48 can modify or modulate the magnetic field, for example, a magnetic field provided by a magnetic field generating device.

[0262] In some embodiments, pattern 48 is arranged in channel 40, for example, in the inner wall of channel 40 and / or on the inner wall of channel 40, but does not overlap with substrate 42, see, for example, as Figure 26The exemplary embodiments are shown. In some embodiments, the pattern 48 is arranged in the channel 40, for example, in the inner wall of the channel 40 and / or on the inner wall of the channel 40, but not in the base 42 and / or on the base 42, see, for example, Figure 26 In some embodiments, pattern 48 is arranged upstream and / or downstream of substrate 42. In some embodiments, pattern 48 overlaps only with substrate 42, and / or only on and / or within substrate 42, see, for example, Figure 27 Alternatively or additionally, pattern 48 may at least partially overlap with substrate 42, and / or may be arranged on and / or in substrate 42, see, for example, Figure 27 or Figure 28 .

[0263] Pattern 48 may be herringbone or may include herringbone patterns. Pattern 48 may be a herringbone pattern or may include herringbone patterns. Pattern 48 may be herringbone-like or may have a herringbone shape.

[0264] Optionally or additionally, the orientation and / or direction of movement of the measuring element can be changed and / or varied by an external force. In some embodiments, the external force can be controlled accordingly and / or can be changed or varied over time and / or locally.

[0265] Alternatively or additionally, the shape of the measuring element 30 may deviate from a sphere, or may not be a perfect sphere (not shown in the figure). Then, during the rolling motion, the movement of the measuring element 30 may vary (during the rolling time), for example, perpendicular to the flow direction, for example, along... Figure 26 , Figure 27 and / or Figure 28 y direction in .

[0266] In some implementations, channel 40 is at least partially or entirely tapered (see example...). Figure 29 (Exemplary implementation shown). For example, channel 40 may be tapered in a direction perpendicular to the flow direction, for example, perpendicular to... Figure 29 In the x-direction. In some embodiments, channel 40 is tapered in the y-direction and / or z-direction. When channel 40 is tapered, the cross-section can vary. When the cross-section increases, the flow velocity may decrease. Channel 40 can be tapered such that its cross-section increases along the flow direction, for example, along the x-direction. Figure 29 In the x-direction. In some embodiments, the measuring element 30 is guided and / or directed to the sensing unit 70 by tapering the channel 40. The channel 40 may be tapered so that the measuring element 30 can be guided and / or directed to the sensor unit 70 and / or sensing elements 72, 74, 76.

[0267] When the channel 40 is tapered, the driving force of the flow 110 on the measuring element 30 can be reduced, and the velocity of the analyte-bound measuring element 30 can be further reduced. Alternatively or additionally, the likelihood of the analyte-bound measuring element 30 being fixed and / or stuck can be increased.

[0268] In a preferred embodiment, device 100 includes a second external force generating device (not shown) configured to generate a second external force. The second external force generating device may be arranged downstream of sensor unit 70, and / or downstream of the most downstream sensing element 74 or the most downstream optical device 78. In some embodiments, the second external force generating device is a second magnetic field generating device or includes a second magnetic field generating device (not shown), and the second external force is magnetic. The second magnetic field generating device may be a permanent magnet or may include a permanent magnet. The second external force generating device may be configured to generate a second external force with varying amplitude and / or spatial orientation. The second external force may vary over time. It may be specified that the second external force generating device is a sorting unit for sorting measurement elements 30. For example, the second external force may be used to sort measurement elements 30, such as those downstream of sensor unit 70. Through sorting, measurement elements 30 may be sorted into different groups. In some embodiments, different kinds / types of analytes are combined with different groups of measurement elements 30. Different groups of measuring elements 30 and / or analytes bound to different groups of measuring elements 30 can be used for further analysis or diagnosis.

[0269] The second external force can be a second magnetic force, but is not limited to this. The second external force can be different from the external force, for example, it can differ in amplitude, power, and / or frequency. It can be specified that the second external force is controlled and / or can vary, for example, with time and / or locally.

[0270] Optionally or additionally, the orientation and / or direction of movement of the measuring element can be changed and / or varied by a second external force. In some embodiments, the second external force can be controlled accordingly, and / or can be changed or varied over time and / or locally.

[0271] Figures 30 and 31 illustrate exemplary measurements performed according to this disclosure. Figure 30a In this context, analyte 10 may be, for example, Her2neu+ EV or may include, for example, Her2neu+ EV, the concentration of which in the sample has been determined according to this disclosure. Figure 30aIn this study, the average velocity difference and average delay of the measuring element 30 were measured using the method and / or apparatus according to this disclosure. In some embodiments, the capture binding elements 35, 36, 37 and / or the surface binding element 42 are anti-Her2neu antibodies or include anti-Her2neu antibodies. The sample may be or may contain a buffer solution, e.g., a suitable chemical. Figure 30a In this context, the average velocity difference corresponds to the velocity change of the measuring element 30. The average delay corresponds, for example, to the corresponding rolling time determined by the sensor unit 70. It can be seen that both the average velocity difference and the average delay monotonically increase with the concentration of analyte 10. Therefore, the concentration can be determined based on the average velocity difference and / or the average delay.

[0272] exist Figure 30b In this context, analyte 10 may be IL-6 or may include IL-6, the concentration of which in the sample has been determined according to this disclosure. Figure 30b In this disclosure, the proportion of the assay elements 30 that are stuck and / or immobilized (relative to the immobilized particles) is shown as a function of the analyte concentration in the sample, where the analyte is, for example, IL-6. The proportion of the assay elements 30 that are stuck and / or immobilized can be determined, for example, by differential counting and / or by detecting the shaking of the assay elements 30. The assay elements 30 that are stuck and / or immobilized have been determined according to this disclosure. In some embodiments, the capture binding elements 35, 36, 37 and / or the surface binding element 42 are or include anti-IL6 antibodies. The sample may be or may contain a buffer, such as, for example, a suitable chemical. It is evident that the proportion of the assay elements 30 that are stuck and / or immobilized increases monotonically with the concentration of the analyte 10. Therefore, the concentration can be determined based on the average velocity difference and / or the average delay. Furthermore, this concentration can be determined over a range of approximately four orders of magnitude and / or orders of magnitude. For example, in Figure 30b The detection range spans from approximately 1 ng / mL to 10,000 ng / mL.

[0273] exist Figure 31a and Figure 31b In this study, flow cytometry was used to determine the proportion of the assay element 30 that was adhered to and / or immobilized (corresponding to the normalized signal). Figure 31a and Figure 31b In this study, optical flow cytometry and magnetic flow cytometry were compared. Figure 31a The concentration of analyte 10 in the buffer solution is displayed, and Figure 31bThe concentrations of analyte 10 in buffer solution (for optical flow cytometry) and blood samples (for magnetic flow cytometry) are displayed. The proportion of assay elements 30 that are stuck and / or immobilized has been determined according to the methods and / or apparatus of this disclosure. The proportion of assay elements 30 that are stuck and / or immobilized can be determined, for example, by differential counting and / or by detecting the shaking of assay elements 30. Figure 31a and Figure 31b In this embodiment, the proportion of the assay elements 30 that are stuck and / or immobilized (relative to the immobilized particles) is shown as a function of the concentration of the analyte (here, biotin) in the sample. In some embodiments, the trapping binder elements 35, 36, 37 and / or the surface binder element 42 are avidin or include avidin. It is evident that the measured proportions of the assay elements 30 determined by two corresponding measurements (i.e., by optical flow cytometry and magnetic flow cytometry) are very close to consistent. In particular, the proportion of the assay elements 30 that are stuck and / or immobilized can also be accurately determined for opaque samples, such as blood (where optical methods may fail). Therefore, magnetic flow cytometry is suitable, for example, for determining the proportion (or number) of the assay elements 30 that are stuck and / or immobilized in opaque samples, and thus also for determining the concentration of the analyte 10. It is evident that the proportion of the assay elements 30 that are stuck and / or immobilized increases monotonically with the concentration of the analyte 10. Therefore, the concentration can be determined from the average velocity difference and / or average delay. Furthermore, this concentration can be determined over a range of approximately four orders of magnitude. For example, in Figure 31a and Figure 31b The detection range spans from approximately 0 mg / mL to 100 ng / mL.

[0274] Figure 31 illustrates an exemplary rate decrease as a function of the analyte concentration in sample 20. In this example, the measuring element 30 has a trapping binder element 35, the substrate 42 has a surface binder element 44, and both the trapping binder element 35 and the surface binder element 44 are configured to bind to the analyte 10 in sample 20. The analyte may include one or more of CD9+ / 63+ / 81+.

[0275] In this example, the rate decrease increases monotonically. Specifically, the rate decrease increases with increasing analyte concentration, such that the determined rate decrease indicates the analyte concentration. In other words, the concentration of analyte 10 in sample 20 can be determined from the rate decrease.

[0276] This disclosure is not limited to the specific exemplary embodiments shown and / or discussed. For some embodiments, only those features given by the independent claims may be needed and / or necessary. It may be specified that a particular embodiment has only some features of some other embodiments. The features disclosed in the claims, description, and drawings may be relevant to the implementation of the invention in any combination thereof.

[0277] List of reference numerals

[0278] 10 Analytes

[0279] 20 samples

[0280] 30 Measuring elements

[0281] 35 (First) Capture binder element

[0282] 36 Second capturing binder element

[0283] 37 Third capture binder element

[0284] 38 Component Test and Analysis Materials

[0285] 40 channels

[0286] 42 Base

[0287] 44 (First) Surface binder element

[0288] 45 Second surface bonding element

[0289] 46 Third surface bonding element

[0290] 47 Surface test analytes

[0291] 48 patterns

[0292] 60 External Forces

[0293] 70 sensor units

[0294] 72 First sensing element

[0295] 74 Second sensing element

[0296] 76 Sensing Elements

[0297] 78 Optical devices

[0298] 80 First distribution function

[0299] 82 Second Distribution Function

[0300] 83 Second Distribution Function

[0301] 100 devices

[0302] 110 Flow

[0303] 112 Sample Flow

[0304] 114 Layered Flow

[0305] 120 External force generating device

[0306] 130 Force.

Claims

1. A method for detecting the concentration of an analyte (10) in a sample (20), the method comprising: A device (100) is provided having a channel (40) having a substrate (42) having surface binder elements (44, 45, 46) configured to bind to the analyte (10) in the sample, and / or the substrate (42) having a surface test analyte (47). The measuring element (30), preferably a magnetic measuring element (30), is mixed into the sample (20), the measuring element (30) having a capture binder element (35, 36, 37) configured to bind to the analyte (10) in the sample, and / or the measuring element (30) having an element test analyte (38) configured to bind to the surface binder element; The sample (20) is allowed to flow through the channel (40); The measuring element (30) is moved to the substrate (42) by an external force (60), preferably a magnetic force; The measuring element (30) is moved along the substrate (42) / on the substrate (42), and the velocity of at least one measuring element (30) is changed by interaction, wherein the interaction includes one or more of the following: (i) The interaction, preferably binding interaction, between at least one surface binder element (44, 45, 46) of the substrate (42), the analyte (10), and at least one trapping binder element (35, 36, 37) of the at least one measuring element (30); (ii) The interaction, preferably binding interaction, between the surface test analyte (47) of the substrate (42) and at least one of the trapping binder elements (35, 36, 37) of the at least one measuring element (30); and / or (iii) The interaction, preferably the binding interaction, between the element test analyte (38) of the at least one measuring element (30) and at least one of the surface binder elements (44, 45, 46) of the substrate (42); as well as The concentration of the analyte (10) is determined using the sensor unit (70) by determining the velocity change of the measuring element (30) along the channel (40), and / or by determining the loss of the measuring element (30) along the channel (40), preferably by determining the loss of the measuring element (30) along the channel (40) through differential counting of the measuring element (30), and / or The binding kinetics, preferably binding constants, between the analyte (10) and the capture binder elements (35, 36, 37) and / or between the analyte (10) and the surface binder elements (44, 45, 46) are determined, wherein the binding kinetics are determined by using the sensor unit (70) to determine the velocity change of the measuring element (30) along the channel (40).

2. The method according to claim 1, wherein moving the measuring element (30) along the substrate (42) / on the substrate (42) comprises rolling the measuring element (30) along the substrate (42) / on the substrate (42).

3. The method according to any one of the preceding claims, wherein the surface test analyte is the same type and / or class as the analyte, and / or wherein the element test analyte is the same type and / or class as the analyte.

4. The method according to any one of the preceding claims, wherein determining the concentration of the analyte (10) includes determining the rolling time between the first sensing element (72, 76) and the second sensing element (74, 76).

5. The method according to claim 4, wherein the first sensing element (72, 76) is disposed upstream of the substrate (42), and wherein the second sensing element (74, 76) is disposed downstream of the substrate (42) and / or downstream of the first sensing element (72, 76).

6. The method according to claim 4 or 5, wherein the first sensing element (72, 76) is a magnetic field sensor or includes a magnetic field sensor, and / or wherein the second sensing element (74, 76) is a magnetic field sensor or includes a magnetic field sensor.

7. The method according to any one of the preceding claims, wherein determining the velocity change comprises determining a first distribution function (80) of the measuring element (30) at a first position in the channel (40) and a second distribution function (82, 83) of the measuring element at a second position in the channel (40). The first distribution function (80) is the distribution function of the time difference between the time point when the corresponding measuring element (30) crosses the first position and the first reference time point, and the second distribution function (82, 83) is the distribution function of the time difference between the time point when the corresponding measuring element crosses the second position and the second reference time point.

8. The method according to claim 7 and further referring to any one of claims 4 to 6, wherein the first position is the position of the first sensing element (72, 76), and / or wherein the second position is the position of the second sensing element (74, 76).

9. The method according to any one of claims 7 or 8, wherein the first distribution function (80) comprises a density function or a histogram, and wherein the second distribution function (82, 83) comprises a density function or a histogram, wherein the velocity change is determined by the time difference between the peak, preferably the maximum value of the second distribution function (82, 83) and the peak, preferably the maximum value of the first distribution function (80).

10. The method according to any one of claims 7 or 8, wherein the first distribution function (80) comprises a cumulative function, and wherein the second distribution function (82, 83) comprises a cumulative function, wherein the velocity change is determined by the time difference between the inflection point of the second distribution function (82, 83) and the inflection point of the first distribution function (80).

11. The method according to any one of claims 7 or 8, wherein the velocity change is determined by a distance metric between the second distribution function (82, 83) and the first distribution function (80) and / or by a shape change of the second distribution function (82, 83) compared to the first distribution function (80).

12. The method of claim 11, wherein the shape change is determined by fitting the first distribution function (80) and the second distribution function (82, 83) to the same candidate distribution function and comparing the corresponding fitting parameters.

13. The method according to claim 12, wherein the candidate distribution function is a normal distribution function, a gamma distribution function, a log-normal distribution function, or a Weibull distribution function.

14. The method according to any one of claims 7 to 13, wherein the first reference time point is equivalent to the second reference time point.

15. The method according to any one of claims 11 to 13, wherein the first reference time point is equivalent to the time point at which the measuring element (30) first crosses the first position, and / or wherein the second reference time point is equivalent to the time point at which the measuring element (30) first crosses the second position.

16. The method according to any one of the preceding claims, wherein determining the concentration of the analyte (10) comprises optical detection, wherein the optical detection preferably comprises capturing the movement and / or position of the measuring element (30) in a spatially and / or temporally resolved field of view (79).

17. The method according to any one of the preceding claims, wherein changing the speed comprises fixing at least one of the measuring elements (30).

18. The method according to any one of the preceding claims, wherein the external force (60) and the flow rate of the sample (20) are adjusted such that at least one of the measuring elements (30) rolls along the substrate (42) / on the substrate (42).

19. The method according to any one of the preceding claims, wherein the external force (60) is adjusted such that the measuring element (30) is pushed away from the substrate (42) and / or sorted, preferably after the concentration of the analyte (10) has been determined.

20. The method according to any one of the preceding claims, wherein in the channel (40), a stratified flow (114) flows between the sample (20, 112) and the substrate (42) to separate the sample (20, 112) from the substrate (42), and wherein the measuring element (30) is moved from the sample (20, 112) to the substrate (42) by the external force (60) through the stratified flow (114).

21. The method according to any one of the preceding claims, wherein at least one of the measuring elements (30) has at least two different capture binding elements (35, 36, 37), wherein the different capture binding elements (35, 36, 37) are configured to bind to different analytes (10), and / or wherein the different capture binding elements (35, 36, 37) are configured to bind to the same analyte (10).

22. The method according to any one of the preceding claims, wherein the capture binder elements (35, 36, 37) of at least one or all of the measuring elements (30) are of the same type and / or kind as at least one or all of the surface binder elements (44, 45, 46).

23. The method according to any one of the preceding claims, wherein at least two of the measuring elements (30) have different sizes, different magnetic moments, different acoustic properties, different electrical properties and / or different optical properties.

24. The method according to any one of the preceding claims, wherein the measuring element (30) comprises at least two sets of measuring elements (30), and each set of measuring elements (30) has the same type and / or kind of capture binder elements (35, 36, 37).

25. The method according to any one of the preceding claims, wherein the substrate (42) has at least two different surface binder elements (44, 45, 46), wherein the different surface binder elements (44, 45, 46) are configured to bind to different analytes (10), wherein preferably the different surface binder elements (44, 45, 46) are grouped into homogeneous groups, preferably separated by sensing elements (76).

26. The method according to any one of the preceding claims, wherein the orientation and / or rolling direction of the measuring element (30) changes when the measuring element (30) moves along the substrate (42) / on the substrate (42), or when the measuring element (30) moves downstream of the substrate (42) or downstream of the sensor unit (70).

27. The method according to any one of the preceding claims, wherein when the measuring element (30) moves downstream of the substrate (42) and / or downstream of the sensor unit (70), a second external force acts on the measuring element (30), the second external force preferably sorting the measuring element (30).

28. An apparatus (100) for detecting the concentration of an analyte (10) in a sample (20), the apparatus (100) comprising: Channel (40), wherein the channel (40) has a substrate (42) having surface binder elements (44, 45, 46) and / or the substrate (40) having surface test analyte (47), the channel (40) being configured such that a sample (20) containing analyte (10) and a measuring element (30) can flow therein, the measuring element (30) having capture binder elements (35, 36, 37) and / or having element test analyte (38); The device (100) is configured such that when the sample (20) containing the analyte (10) and the measuring element (30) flows through the channel, an external force (60) moves the measuring element (30) to the substrate (42), and the measuring element (30) moves along the substrate (42) / on the substrate (42). The device is further configured such that when the measuring element (30) moves along the substrate (42) or on the substrate (42), the velocity of at least one measuring element is changed by an interaction, the interaction including one or more of the following: (i) The interaction, preferably binding interaction, between at least one surface binder element (44, 45, 46) of the substrate (42), the analyte (10), and at least one trapping binder element (35, 36, 37) of the at least one measuring element (30); (ii) The interaction, preferably binding interaction, between the surface test analyte of the substrate and at least one of the trapping binder elements (35, 36, 37) of the at least one measuring element (30); and / or (iii) The interaction, preferably a binding interaction, between the test analyte of the at least one measuring element and at least one of the surface binder elements (44, 45, 46); and The device further includes a sensor unit (70) configured to determine the velocity variation of the measuring element (30) along the channel (40), and / or determine the loss of the measuring element (30) along the channel (40), preferably by differential counting of the measuring element (30), and / or determine the binding kinetics, preferably binding constants, between the analyte (10) and the trapping binder elements (35, 36, 37) and / or between the analyte (10) and the surface binder elements (44, 45, 46).

29. The apparatus (100) according to claim 28, wherein the apparatus (100) includes an external force generating device (120) configured to generate the external force (60).

30. The apparatus (100) according to claim 29, wherein the external force generating device (120) is a magnetic field generating device or includes a magnetic field generating device, and the external force (60) is a magnetic force, wherein preferably the magnetic field generating device is a permanent magnet or includes a permanent magnet.

31. The apparatus (100) according to any one of claims 28 to 30, wherein the sensor unit (70) includes a first sensing element (72, 76) and a second sensing element (74, 76), wherein the sensor unit (70) is configured to determine the concentration of the analyte (10) by determining the rolling time between the first sensing element (72, 76) and the second sensing element (74, 76).

32. The apparatus (100) according to claim 31, wherein the first sensing element (72, 76) is disposed upstream of the substrate (42), wherein preferably the second sensing element (74, 76) is disposed downstream of the first sensing element (72, 76) and / or the substrate (42), wherein preferably the first sensing element (72, 76) and / or the second sensing element (74, 76) respectively comprise a magnetic field sensor or a magnetic field sensor.

33. The apparatus (100) according to any one of claims 28 to 32, wherein the sensor unit (70) is configured to determine the velocity change by determining a first distribution function (80) of the measuring element (30) at a first position in the channel (40) and a second distribution function (82, 83) of the measuring element (30) at a second position in the channel (40). The first distribution function (80) is the distribution function of the time difference between the time point when the corresponding measuring element (30) crosses the first position and the first reference time point, and the second distribution function (82, 83) is the distribution function of the time difference between the time point when the corresponding measuring element (30) crosses the second position and the second reference time point.

34. The apparatus (100) of claim 33 and further referring to claim 32, wherein the first position is the position of the first sensing element (72, 76), and / or wherein the second position is the position of the second sensing element (74, 76).

35. The apparatus (100) according to claim 33 or 34, wherein the first distribution function (80) comprises a density function or a histogram, and wherein the second distribution function (82, 83) comprises a density function or a histogram, wherein the velocity change is determined by the time difference between the peak, preferably the maximum value of the second distribution function (82, 83) and the peak, preferably the maximum value of the first distribution function (80).

36. The apparatus (100) according to claim 33 or 34, wherein the first distribution function (80) includes a cumulative function, and wherein the second distribution function (82, 83) includes a cumulative function, wherein the velocity change is determined by the time difference between the inflection point of the second distribution function (82, 83) and the inflection point of the first distribution function (80).

37. The apparatus (100) according to claim 33 or 34, wherein the velocity change is determined by the distance between the second distribution function (82, 83) and the first distribution function (80), and / or by the shape change of the second distribution function (82, 83) compared to the first distribution function (80).

38. The apparatus (100) according to any one of claims 28 to 37, wherein the sensor unit (70) is an optical device (78) or includes an optical device (78), wherein the optical device (78) is preferably configured to determine the concentration of the analyte (10) by capturing the movement and / or position of the measuring element (30) in at least one field of view (79) with spatial resolution and / or temporal resolution.

39. The apparatus (100) according to any one of claims 28 to 38, wherein the substrate (42) has at least two different surface binder elements (44, 45, 46), wherein the different surface binder elements (44, 45, 46) are configured to bind to different analytes (10), wherein preferably the different surface binder elements (44, 45, 46) are grouped into homogeneous groups, preferably separated by sensing elements (76).

40. The apparatus (100) according to any one of claims 28 to 39, wherein the apparatus is configured to change the orientation and / or rolling direction of the measuring element (30) when the measuring element (30) rolls along the substrate (42) / on the substrate (42) or downstream of the substrate (42) or downstream of the sensor unit (70), and / or before the measuring element (30) rolls along the substrate (42) / on the substrate (42) or downstream of the substrate (42) or downstream of the sensor unit (70).

41. The device (100) according to claim 40, wherein the channel (40) and / or the substrate (42) have mechanical and / or magnetic patterns (48), preferably mechanical and / or magnetic herringbone patterns.

42. The device (100) according to any one of claims 28 to 41, wherein the channel (40) is tapered.

43. The device (100) according to any one of claims 28 to 42, wherein the device (100) includes a second external force generating device configured to generate a second external force, wherein preferably the second external force generating device is arranged at a position along the substrate (42) or downstream of the sensor unit (70).

44. The apparatus (100) according to claim 43, wherein the second external force generating device is a second magnetic field generating device or includes a second magnetic field generating device, and the second external force is a magnetic force, wherein preferably the second magnetic field generating device is a permanent magnet or includes a permanent magnet, and / or wherein preferably the second external force generating device is configured to generate the second external force with varying amplitude and / or spatial direction.

45. The apparatus (100) according to claim 43 or 44, wherein the second external force generating device is configured to sort the measuring element (30).

Images