Analysis device

Abstract

This analysis device detects a to-be-inspected substance in a sample while performing component separation by means of electrophoresis on the sample supplied to a flow channel of a measurement chip provided with the flow channel. The flow channel of the measurement chip includes a main flow channel and a plurality of branch flow channels branched from the main flow channel. The analysis device comprises: an electrophoresis mechanism that subjects the to-be-inspected substance to electrophoresis; and a detection unit that detects the to-be-inspected substance, wherein the electrophoresis mechanism forms, in the main flow channel, a concentrated layer in which the to-be-inspected substance is concentrated, branches the concentrated layer into the plurality of branch flow channels, and then subjects the to-be-inspected substance to gel electrophoresis in the branch flow channels. The detection unit detects the to-be-inspected substance from each of the plurality of branch flow channels.

Description

Analyzer

[0001] This disclosure relates to an analytical instrument.

[0002] Conventionally, analytical devices utilizing electrophoresis with microfluidic chips are known. For example, a technique is known in which isokinetic electrophoresis is performed to concentrate the sample and form a concentrated layer, and then the process is switched from isokinetic electrophoresis to capillary electrophoresis to separate specific components contained in the concentrated layer. Japanese Patent Publication No. 2003-536058 discloses a microfluidic chip that can also be used in electrophoresis devices. Japanese Patent Publication No. 2003-536058 discloses a microfluidic chip equipped with multiple channels that enable the simultaneous detection of multiple targets.

[0003] Japanese Patent Publication No. 2003-536058 does not disclose a specific method for applying a microfluidic chip with multiple channels to an electrophoretic analytical device.

[0004] This disclosure aims to provide an analytical device that utilizes electrophoresis and can perform multiple tests in parallel.

[0005] To achieve the above objective, an analytical apparatus according to a first aspect of the technology of this disclosure is an analytical apparatus for detecting a target substance in a sample while performing component separation by electrophoresis on a sample supplied to a channel of a measuring chip equipped with a channel, wherein the channel of the measuring chip includes a main channel and a plurality of branch channels branching off from the main channel, and comprises an electrophoresis mechanism for electrophoresing the target substance and a detection unit for detecting the target substance, wherein the electrophoresis mechanism forms a concentrated layer in the main channel by concentrating the target substance, branches the concentrated layer into a plurality of branch channels, performs gel electrophoresis on the target substance in the branch channels, and the detection unit detects the target substance from each of the plurality of branch channels.

[0006] In the first embodiment, the electrophoresis mechanism preferably forms a concentrated layer by performing constant-rate electrophoresis on the substance to be tested in the main channel.

[0007] In the first embodiment, the electrophoresis mechanism preferably has a detection function to detect the position of the concentrated layer in the main channel, and switches the channel end to which voltage or current is applied among a plurality of channel ends in the channel according to the position of the concentrated layer, thereby initiating gel electrophoresis in the branched channel.

[0008] In the first embodiment, it is preferable to have a reagent filling mechanism that introduces reagents containing different components into at least two of the branched channels of a plurality of branched channels.

[0009] In the first embodiment, the reagent filling mechanism preferably introduces the reagent into the branched channel by dispensing the reagent into the end of the branched channel and applying pressure.

[0010] In the first embodiment, it is preferable to include different molecules as distinct components that specifically bind to different structures of the substance to be tested.

[0011] In the first embodiment, it is preferable that the substance to be tested is a specific protein, and that the molecules being tested are molecules that specifically bind to different sugar chains for detecting sugar chain mutations in the protein.

[0012] In the first embodiment, it is preferable that the detection unit is a photodetection unit that detects an optical signal corresponding to the substance to be inspected.

[0013] In the first embodiment, it is preferable that the optical signal is fluorescence.

[0014] A second embodiment of the technology of the present disclosure is an analytical apparatus for detecting a target substance in a sample while performing component separation by electrophoresis on a sample supplied to a channel of a measuring chip equipped with a channel, wherein the channel of the measuring chip includes a main channel and a plurality of branch channels branching off from the main channel, and comprises an electrophoresis mechanism for electrophoresing the target substance in the main channel and the plurality of branch channels, and a single photodetection unit for detecting the target substance, wherein the electrophoresis mechanism branches the target substance from the main channel to the plurality of branch channels, shifts the time it takes for the target substance to reach the detection site in each of the plurality of branch channels, and the single detection unit sequentially detects the target substance from the detection site in each of the plurality of branch channels.

[0015] In the second embodiment, the electrophoresis mechanism preferably shifts the time it takes for the substance to be tested to reach the detection site in each branch channel by sequentially applying a first current, which has a relatively larger absolute value than the other branch channels where detection has not been completed, to each branch channel, and by applying a voltage or current to the channels that prevents the substance to be tested from flowing back into the main channel from the other branch channels where detection has not been completed.

[0016] In a second embodiment, the electrophoresis mechanism preferably applies a voltage or current to the channels through which a second current, which has a relatively smaller absolute value than the first current, flows in the same direction as the first current, while a first current is flowing through one branch channel.

[0017] In the second embodiment, it is preferable to have a detection unit movement mechanism that moves one detection unit between each detection site in a plurality of branched flow channels.

[0018] In the second embodiment, it is preferable to have a measurement chip moving mechanism that moves the measurement chip so that each detection point of the multiple branched channels is sequentially positioned at a detection position by a single photodetector.

[0019] In the second embodiment, it is preferable that the detection unit is a photodetection unit that detects an optical signal corresponding to the substance to be inspected.

[0020] In the second embodiment, it is preferable that the optical signal is fluorescence.

[0021] In a second embodiment, the electrophoresis mechanism preferably forms a concentrated layer in the main channel where the substance to be tested is concentrated, branches the concentrated layer into a plurality of branched channels, and then performs gel electrophoresis on the substance to be tested in the branched channels.

[0022] In a second embodiment, the electrophoresis mechanism preferably forms a concentration layer by performing constant-velocity electrophoresis on the substance to be tested in the main channel.

[0023] In a second embodiment, the electrophoresis mechanism preferably has a detection function to detect the position of the concentrated layer in the main channel, and switches the channel end to which voltage or current is applied among a plurality of channel ends in the channel according to the position of the concentrated layer, thereby initiating gel electrophoresis in the branched channel.

[0024] According to the technology disclosed herein, it is possible to perform multiple tests in parallel using electrophoresis.

[0025] This is a schematic diagram of the analytical apparatus of the first embodiment. This is a schematic diagram showing an example of the flow path of the measuring chip of the first embodiment. This is a block diagram showing an example of the hardware configuration of the control device of the embodiment. This is a process diagram of the analytical method in the analytical apparatus of the first embodiment (part 1). This is a process diagram of the analytical method in the analytical apparatus of the first embodiment (part 2). This is an explanatory diagram of application example 1. This is an explanatory diagram of application example 2. This is an explanatory diagram of application example 3. This is a schematic diagram of the analytical apparatus of the second embodiment. This is a schematic diagram showing an example of the flow path of the measuring chip of the second embodiment. This is a schematic diagram showing the reagent-filled state of the measuring chip of the second embodiment. This is a schematic diagram of the analytical apparatus of the third embodiment. This is a process diagram of the analytical method in the analytical apparatus of the third embodiment (part 1). This is a process diagram of the analytical method in the analytical apparatus of the third embodiment (part 2). This is an explanatory diagram of modified example 1 of the analytical method. This is an explanatory diagram of modified example 2 of the analytical method. This is a schematic diagram of the flow path of the modified example.

[0026] Hereinafter, an analyzer according to an embodiment of the present disclosure will be described with reference to the drawings. In each drawing, components shown using the same reference numeral mean the same component. Unless otherwise specifically stated in the specification, each component is not limited to one, and a plurality of components may be present.

[0027] "Analyzer of First Embodiment" FIG. 1 is a schematic diagram showing a schematic configuration of an analyzer 100 according to a first embodiment of the technology of the present disclosure. The present embodiment is one form of the analyzer according to the first aspect. The analyzer 100 detects a test target substance A (see FIG. 2) in a sample while performing component separation by electrophoresis on the sample (specimen) supplied to the flow path of the measurement chip 10 having the flow path 20. As an example, the sample is a biological sample such as blood collected from a living body. The analyzer 100 includes a support section (not shown) that supports the measurement chip 10, an electrophoresis mechanism 12, detection units 16A and 16B, and a control device 18. Here, "detecting the test target substance A" includes detecting a signal derived from the test target substance A itself, a complex containing the test target substance A, or a labeled molecule that specifically binds to the test target substance A.

[0028] The measurement chip 10 is a microchannel device provided with a microchannel as the channel 20, and as an example, is a measurement chip for μTAS (Micro-Total Analysis System). The measurement chip 10 is used by being loaded into the analyzer 100. Here, the microchannel refers to a channel on the micrometer (μm) scale. More specifically, a microchannel refers to a channel in which the width and height of the cross-section of the channel through which a fluid passes are less than 1 mm, and are mainly on the order of several tens of micrometers to several hundreds of micrometers. The measurement chip 10 is a single-use type that is used once for one sample.

[0029] In FIG. 1, the flow path 20 of the measurement chip 10 is shown schematically. FIG. 2 is an extracted view showing the flow path of the measurement chip 10.

[0030] The flow path 20 of the measurement chip 10 includes a main flow path 21 and a plurality of branched flow paths branched from the main flow path 21 (here, a first branched flow path 22 and a second branched flow path 23). Further, the flow path 20 includes four flow path ends, and wells 26a to 26d are respectively provided at the flow path ends. In this specification, the term "flow path end" refers to an end portion of the flow path 20, and means a portion to which only the main flow path 21 or the branched flow paths 22 and 23 are connected in one direction. The well 26a constitutes a flow path end of the main flow path 21. The well 26b constitutes a flow path end of the first branched flow path 22, and the well 26c constitutes a flow path end of the second branched flow path 23. Further, the well 26d constitutes a flow path end of a connection path 24 connected to the main flow path 21. Hereinafter, the location where the main flow path 21 branches into the first branched flow path 22 and the second branched flow path 23 is referred to as a branch point 29.

[0031] When the electrophoresis mechanism 12 applies voltage or current to the flow path 20, electrodes are inserted into the four flow path ends, which are the wells 26a to 26d. An electrophoresis flow path is formed by the main flow path 21, the first branched flow path 22, and the second branched flow path 23. The well 26a side, which is the flow path end of the main flow path 21, is referred to as the "upstream side", and the well 26b and 26c sides, which are the flow path ends of the branched flow paths 22 and 23, are referred to as the "downstream side".

[0032] The electrophoresis flow path has a concentration region 30 and a component separation region 32 from the upstream side toward the downstream side. The concentration region 30 is a region for concentrating a test substance (for example, a tumor marker) A using, for example, an immune reaction and isotachophoresis (ITP: IsoTachoPhoresis). In the example shown in FIG. 1, the concentration region 30 is provided from the upstream side of the main flow path 21 to the branch point 29 between the main flow path 21 and the branched flow paths 22 and 23. The component separation region 32 is a region for separating the test substance A from other components using capillary gel electrophoresis (hereinafter simply referred to as gel electrophoresis). The first branched flow path 22 and the second branched flow path 23 each constitute the component separation region 32.

[0033] Further, the first branched flow path 22 and the second branched flow path 23, which serve as the component separation region 32, respectively include detection sites 34A and 34B for detecting the test substance A.

[0034] Although not shown in Figure 1, reagents S1 to S5 are pre-filled in the channel 20 of the measurement chip 10 (see Figure 2). For example, as shown in Figure 2, the concentration region 30 is filled with reagents S1 to S5 containing buffer solutions adjusted so that the conductivity increases in steps from upstream to downstream. Reagent S1 has the lowest conductivity, and reagent S5 has the highest conductivity. Reagent S2 contains a buffer solution and a first-labeled antibody D having a specific polarity charge (e.g., a DNA (deoxyribonucleic acid) labeled antibody). Reagent S3 contains a buffer solution, a sample, and a second-labeled antibody B (e.g., a fluorescently labeled antibody). If the sample contains the substance to be tested A, an immunocomplex C is formed in reagent S3 by binding the substance to be tested A and the second-labeled antibody B. The first branch channel 22 and the second branch channel 23, which constitute the component separation region 32, are filled with reagent S5. Reagent S5 contains a gel for gel electrophoresis. In this embodiment, the second labeled antibody B is a fluorescently labeled antibody.

[0035] The electrophoresis mechanism 12 applies a voltage between predetermined channel ends to electrophores the substance A to be tested. The electrophoresis mechanism 12 forms a concentrated layer F in the main channel 21 by concentrating the substance A to be tested, then branches the concentrated layer F into a plurality of branch channels 22 and 23, and then performs gel electrophoresis on the substance A to be tested in the branch channels 22 and 23. The electrophoresis mechanism 12 includes a power supply for applying voltage or current to the channel 20, a plurality of electrodes connected to the power supply for insertion into the wells 26a to 26d described above, and a control unit. The control unit includes a processor and controls the power supply to control which region in the channel 20 to apply voltage or current to (i.e., between which wells to apply voltage), the timing of voltage application, and the magnitude of the voltage. Some or all of the functions of the control unit may be provided in the control device 18.

[0036] In this embodiment, the electrophoresis mechanism 12 forms a concentrated layer F by performing constant-velocity electrophoresis on the substance A to be tested in the main channel 21. The electrophoresis mechanism 12 also has a detection function to detect the position of the concentrated layer F in the main channel 21. The detection function is implemented by the control unit of the electrophoresis mechanism 12. The electrophoresis mechanism 12 also changes the region to which voltage or current is applied in the channel 20 according to the position of the concentrated layer F, that is, it switches which channel end 26a to 26d of the channel 20 to which voltage or current is applied, and starts gel electrophoresis of the substance A to be tested in the branch channels 22 and 23. More specifically, the electrophoresis mechanism 12 detects the position of the concentrated layer F and applies voltage or current for gel electrophoresis to each branch channel 22 and 23 at the timing when the concentrated layer F in the main channel 21 branches into each branch channel 22 and 23.

[0037] Detection units 16A and 16B are used to detect the substance A to be tested, for example, by optically detecting immune complexes. Detection unit 16A is positioned opposite the detection site 34A in the first branch channel 22. Detection unit 16B is positioned opposite the detection site 34B in the second branch channel 23.

[0038] Detection units 16A and 16B are photodetection units that detect an optical signal (in this case, fluorescence) corresponding to the substance A to be inspected. The photodetection unit includes an objective lens, an excitation light source, and a photodetector. The excitation light source is a laser diode (LD) or light-emitting diode (LED), etc., that emits excitation light. The photodetector is a photodiode (PD) or photomultiplier tube (PMT), etc., for detecting fluorescence generated by excitation by the excitation light. The excitation light is focused by the objective lens and irradiated onto the detection site, and the fluorescence emitted from the detection site is focused by the objective lens and received by the photodetector. Detection units 16A and 16B output a detection signal corresponding to the received fluorescence to the control device 18.

[0039] The control device 18 has the function of performing overall control related to electrophoresis. Specifically, it controls the electrophoresis mechanism 12 and the detection units 16A and 16B. The control device 18 controls the start of detection by the detection units 16A and 16B according to the timing of gel electrophoresis by the electrophoresis mechanism 12. The control device 18 also receives detection signals from the detection units 16A and 16B and outputs the received results.

[0040] Figure 3 shows a block diagram illustrating an example of the hardware configuration of the control device 18. As shown in Figure 3, the control device 18 includes a processor 80 such as a CPU (Central Processing Unit), memory 82, an I / F (Interface) unit 83, a storage unit 84, a display 86, and an input device 88. The processor 80, memory 82, I / F unit 83, storage unit 84, display 86, and input device 88 are connected to each other via a bus 89 such as a system bus or control bus, enabling the exchange of various types of information.

[0041] The processor 80 reads various programs, including the control program 85 stored in the storage unit 84, into the memory 82 and executes processing according to the read programs. In this way, the processor 80 performs control related to electrophoresis. The memory 82 is a work memory for which the processor 80 executes processing.

[0042] The control program 85 executed by the processor 80 is stored in the storage unit 84. Specific examples of the storage unit 84 include HDDs (Hard Disk Drives) and SSDs (Solid State Drives).

[0043] The I / F unit 83 communicates various information with the electrophoresis mechanism 12 and detection units 16A and 16B via wireless or wired communication. The display 86 and input device 88 function as a user interface. The display 86 provides the user with various information regarding the analysis of the sample. The display 86 is not particularly limited and can be an LCD monitor, LED monitor, etc. The input device 88 is operated by the user to input various instructions regarding the operation of the analyzer 100, such as setting analysis conditions, instructing the start of measurement, instructing the display switching of measurement results, etc. The input device 88 is not particularly limited and can be an example of a keyboard, stylus, mouse, etc. The control device 18 employs a touch panel display that integrates the display 86 and the input device 88.

[0044] The method for analyzing a sample using the analytical apparatus configured as described above will be explained with reference to Figures 4 and 5.

[0045] As shown in Figure 2, the flow path 20 of the measurement chip 10 is filled with reagents S1 to S5, which are adjusted so that the conductivity increases in steps from the upstream side to the downstream side. As previously described, reagent S2 contains the first labeled antibody D, and reagent S3 contains the sample and the second labeled antibody B. In this example, the sample contains the antigen A, which is the substance to be tested, and reagent S3 contains an immunocomplex C in which the substance to be tested A is bound to the second labeled antibody B. Here, the first labeled antibody D is assumed to have a negative charge as a charge of a specific polarity. If the charge of a specific polarity is a positive charge, the cathode and anode should be reversed in the following explanation.

[0046] As shown in Step 1 of Figure 4, the electrophoresis mechanism 12 applies a voltage between wells 26a and 26b, and between wells 26a and 26c, with the electrode inserted in well 26a as the cathode (-) and the electrodes inserted in wells 26b and 26c as the anodes (+). This allows isokinetic electrophoresis to be performed in the concentration region 30 of the main channel 21. The negatively charged first-labeled antibody D moves towards the anode side while being concentrated, and the antigen-antibody reaction causes the target substance A and the first-labeled antibody D to bind, forming a sandwich-type immune complex E. That is, immune complex C and the first-labeled antibody D bind to form immune complex E. As a result, the target substance A is gradually concentrated.

[0047] As shown in Step 2 of Figure 4, the uncharged second-labeled antibody B does not move, while the first-labeled antibody D and the immune complex E containing the first-labeled antibody D move towards the anode, resulting in B / F (Bound / Free) separation and the gradual formation of the enriched layer F (see Step 3).

[0048] As shown in Step 3 of Figure 4, the electrophoresis mechanism 12 controls the power supply to apply a voltage Vj to the electrode inserted in the well 26d during isokinetic electrophoresis so that no current flows through the connection channel 24. The state in which no current flows through the connection channel 24 is when the potential of the well 26d is equal to the potential of the connection point 33 where the connection channel 24 connects to the main channel 21. Therefore, by monitoring the voltage Vj at which no current flows through the connection channel 24, the potential in the connection channel 24 can be monitored. Due to isokinetic electrophoresis, the reagent S1 with high resistance (low conductivity) gradually moves from the upstream side to the downstream side, causing the potential at the connection point 33 to gradually change. By monitoring this change in the potential at the connection point 33, the position of the concentrated layer F at the connection point 33 can be detected.

[0049] Furthermore, the concentrated layer F moves downstream, and at the branching point 29, the concentrated layer F branches into the first branch channel 22 and the second branch channel 23 (see Step 4 in Figure 5). Subsequently, as shown in Step 4 in Figure 5, the cathode is switched from well 26a to well 26d, thereby switching from isokinetic electrophoresis to gel electrophoresis. The electrophoresis mechanism 12 detects the position of the concentrated layer F by monitoring the change in potential at the connection point 33, and switches from isokinetic electrophoresis to gel electrophoresis at the timing when the concentrated layer F reaches the branching point 29 and branches into the first branch channel 22 and the second branch channel 23. The electrophoresis mechanism 12 switches the cathode from well 26a to well 26d and applies a voltage between well 26d and well 26b, and between well 26d and well 26c. As a result, gel electrophoresis of the substance A to be tested is started in the first branch channel 22 and the second branch channel 23, respectively. In gel electrophoresis in the first branch channel 22 and the second branch channel 23, component separation is performed by the molecular sieving effect.

[0050] Then, as shown in Step 5 of Figure 5, the target substance A that has reached detection sites 34A and 34B is detected by detection units 16A and 16B, respectively. Specifically, detection unit 16A irradiates detection site 34A with excitation light and detects the fluorescence generated from detection site 34A due to the irradiation of excitation light. Similarly, detection unit 16B irradiates detection site 34B with excitation light and detects the fluorescence generated from detection site 34B due to the irradiation of excitation light. The fluorescence is generated from the second-labeled antibody B contained in the immune complex E that passes through detection site 34A or detection site 34B. Detection units 16A and 16B acquire the time change of fluorescence intensity. Due to the molecular sieving effect, the immune complex E in the enriched layer branched into the first branch channel 22 passes through detection site 34A near the peak time in the time change of fluorescence intensity. Similarly, due to the molecular sieving effect, the immune complex E in the enriched layer branched into the second branch channel 23 passes through detection site 34B near the peak time in the time change of fluorescence intensity. The target substance A can be quantified from the peaks in the time-dependent changes in fluorescence intensity obtained by detection units 16A and 16B.

[0051] As described above, the analyzer 100 of this embodiment has a measurement chip 10 equipped with multiple (in this case, two) branched channels 22 and 23, so that the substance A to be tested can be detected in each of the branched channels 22 and 23. Since it is equipped with multiple branched channels 22 and 23 and multiple tests can be performed in parallel, the accuracy of the tests can be improved and at least one of multiple items can be tested, thereby improving the diagnostic performance of the analyzer 100.

[0052] As in this embodiment, when detecting the same target substance A in branched channels 22 and 23, the detection results can be integrated and evaluated, thereby improving detection accuracy. In other words, by performing measurements using a measurement chip 10 equipped with multiple branched channels 22 and 23, higher accuracy detection results can be obtained compared to when a measurement chip with only one channel in the component separation region 32 is used. With a measurement chip with only one channel in the component separation region 32, improving accuracy required either lengthening the channel or performing the same test multiple times using multiple measurement chips. Both of these have the disadvantage of increasing measurement time, but as in this embodiment, by performing measurements using a measurement chip 10 equipped with multiple branched channels 22 and 23, accuracy can be improved without increasing the inspection time.

[0053] Furthermore, in the analytical apparatus 100 of this embodiment, the electrophoresis mechanism 12 forms a concentrated layer F in the main channel 21 by concentrating the substance A to be tested, and branches the concentrated layer F into a plurality of branched channels 22 and 23. By forming the concentrated layer F before starting gel electrophoresis, the substance A to be tested can be subjected to gel electrophoresis in a high-density state, which enhances the molecular sieving effect and improves detection accuracy.

[0054] In the above description, the concentrated layer F is formed in the main channel 21 using isokinetic electrophoresis. However, any method other than isokinetic electrophoresis can be used to form the concentrated layer F. Methods for forming the concentrated layer include the Field amplified stacking (FAS) method, Micelle sweeping method, Perm-selective method, and Solid Phase Extraction (SPE) method. For details on these concentration methods, please refer to Sachio Yamamoto, et al., "Microchip Electrophoresis and Sample Concentration," Biophysical Chemistry, 2008; 52: 127.

[0055] In the analytical apparatus 100 of this embodiment, the electrophoresis mechanism 12 has a detection function to detect the position of the concentrated layer F in the main channel 21. Depending on the position of the concentrated layer F, it switches the channel end to which voltage or current is applied among the multiple channel ends 26a to 26d of the channel 20, thereby switching to gel electrophoresis of the substance A to be tested in the branched channels 22 and 23. By detecting the position of the concentrated layer F, it is possible to switch to gel electrophoresis only after the concentrated layer F has reliably branched into the branched channels 22 and 23.

[0056] In this embodiment, the position of the enriched layer F is detected by monitoring the change in potential at the connection point 33, but the method for detecting the position of the enriched layer F is not limited to this embodiment. Any method that can directly or indirectly detect the time at which the enriched layer F reaches the branching point 29 is acceptable, and the analyzer 100 may be configured to detect the position of the enriched layer F by other electrical, optical, or magnetic methods.

[0057] In the above embodiment, the case in which the same inspection is performed in multiple branched channels 22 and 23 has been described, but it is also possible to perform different inspections in multiple branched channels 22 and 23. Here, different inspections may detect different target substances, or they may detect multiple structural displacements of a single target substance.

[0058] By introducing reagents containing different components into the two branched channels 22 and 23, various tests can be performed. By making the pH of the reagents packed into multiple branched channels 22 and 23 different, differences in isoelectric points can be detected. By making the sizes of the electrophoretic gels in the reagents packed into multiple branched channels 22 and 23 different, differences in molecular weight can be detected. By adding different lectins to the reagents packed into multiple branched channels 22 and 23, differences in carbohydrate chain structure can be detected when the substance to be detected is a protein. Furthermore, structural differences can also be detected by adding multiple antibodies that recognize a specific structure and detecting different antibodies in multiple branched channels 22 and 23.

[0059] An example of an application using a measurement chip 10 having multiple branched channels 22 and 23 will be described.

[0060] (Application Example 1) As an example, let's explain the case where AFP (alpha-fetoprotein), a tumor marker for hepatocellular carcinoma, is the substance to be tested. AFP is a protein. Differences in the sugar chain structure of AFP can be detected by the presence or absence of lectin binding. AFP-L1 has no lectin affinity and does not bind to lectins. On the other hand, AFP-L3 has lectin affinity and binds to lectins. It is known that AFP-L3 increases in hepatocellular carcinoma, and the ratio of AFP-L3 to total AFP is used to diagnose hepatocellular carcinoma.

[0061] As shown in Figure 6, lectin L is added to reagent S5 which is filled into the first branch channel 22 of the measurement chip 10. On the other hand, lectin L is not added to reagent S5 which is filled into the second branch channel 23. The fluorescently labeled AFP is concentrated in the concentration region 30 by ITP or the like and branched into the first branch channel 22 and the second branch channel 23. In the first branch channel 22, AFP is fractionated into AFP-L1 and AFP-L3 based on its reactivity with lectin. As a result, at detection site 34A, as shown in graph G1 in Figure 6, a peak of AFP-L3, which has affinity for lectin, and a peak of AFP-L1, which has no affinity for AFP-L3, are detected. On the other hand, at detection site 34B, since lectin L is not present and AFP does not undergo fractionation due to differences in sugar chain structure, a peak of AFP is detected as shown in graph G2 in Figure 6. The AFP peak in graph G2 represents AFP that is not bound to lectin L, and therefore it is detected at the same time as the AFP-L1 peak in graph G1. Consequently, by referring to graph G2, the AFP-L3 peak in graph G1 can be extracted.

[0062] In graph G1 in Figure 6, the peaks for AFP-L1 and AFP-L3 are clearly visible. However, in actual detection, due to the versatility of the sample, it is very rare but difficult to separate the two. As in this example, by acquiring graph G2 in parallel with graph G1, graph G2 can be referenced, allowing for accurate separation of the AFP-L3 peak from graph G1.

[0063] This method allows for more accurate evaluation of the binding affinity to lectin L than conventional methods. By providing multiple branched channels 22 and 23, and introducing reagents with different presences of lectin L into each branched channel 22 and 23, the accuracy of detecting specific glycosylation mutations (in this case, AFP-L3) in a protein (in this case, AFP) can be improved.

[0064] (Application Example 2) Alternatively, reagents containing different molecules that specifically bind to different structures of the substance A to be tested may be introduced into multiple branched channels 22 and 23 as different components. Specifically, if the substance A to be tested is a specific protein, the different molecules are molecules that specifically bind to different sugar chains for detecting sugar chain mutations in the protein.

[0065] Specifically, if the substance to be tested A is a specific protein, and protein P is one that undergoes multiple glycosylation mutations, this method can be used to detect multiple glycosylation mutations.

[0066] As shown in Figure 7, lectin L1 is added to reagent S5 which is filled into the first branch channel 22 of the measurement chip 10, and lectin L2 is added to reagent S5 which is filled into the second branch channel 23. Lectin L1 specifically binds to the first glycan structure of protein P, and lectin L2 specifically binds to the second glycan structure of protein P.

[0067] The fluorescently labeled protein P is concentrated in the enrichment region 30 by ITP or the like, and then branched into the first branch channel 22 and the second branch channel 23.

[0068] In the first branch channel 22, protein P is fractionated into those having a first glycan structure that is compatible with lectin L1 and those not having a first glycan structure. As a result, at the detection site 34A, as shown in graph G11 in Figure 7, a peak from protein P without a first glycan structure and a peak from protein P having a first glycan structure that is compatible with lectin L1 (L1 affinity peak in graph G11) are detected.

[0069] On the other hand, in the second branch channel 23, protein P is fractionated into those having a second glycan structure that is compatible with lectin L2 and those not having a second glycan structure. As a result, at the detection site 34B, as shown in graph G12 in Figure 7, a peak from protein P without a second glycan structure and a peak from protein P having a second glycan structure that is compatible with lectin L2 (L2 affinity peak in graph G12) are detected.

[0070] In this way, the presence or absence of multiple glycan mutations in a single target substance A (in this case, protein P) can be simultaneously detected in the first branch channel 22 and the second branch channel 23. That is, multiple glycan mutations in a single target substance A can be detected in a single measurement.

[0071] (Application Example 3) It can also be used to detect multiple target substances A in a sample. Here, the multiple target substances A are protein PA and protein PB.

[0072] As shown in Figure 8, lectin LA is added to reagent S5 which is filled into the first branch channel 22 of the measurement chip 10, and lectin LB is added to reagent S5 which is filled into the second branch channel 23. Lectin LA specifically binds to a specific sugar chain structure of protein PA but does not bind to protein PB. On the other hand, lectin LB specifically binds to a specific sugar chain structure of protein PB but does not bind to protein PA. In addition, the fluorescent labels mixed with the sample include a first fluorescent label that binds to protein PA and a second fluorescent label that binds to protein PB. The first and second fluorescent labels emit fluorescence at different wavelengths.

[0073] In this case, the first detection unit 16A and the second detection unit 16B may be, for example, equipped with a multicolor fluorescence detector. Alternatively, the first detection unit 16A may be equipped with a bandpass filter for detecting only the fluorescence of the first fluorescent label, and the second detection unit 16B may be equipped with a bandpass filter for detecting only the fluorescence of the second fluorescent label. A multicolor fluorescence detector is preferable from the viewpoint of cost and device size.

[0074] Protein PA, which has been given the first fluorescent label, and protein PB, which has been given the second fluorescent label, are concentrated in the enrichment region 30 by ITP or the like, and then branched into the first branch channel 22 and the second branch channel 23. Both protein PA and protein PB are introduced into both the first branch channel 22 and the second branch channel 23.

[0075] In the first branch channel 22, protein PA is fractionated into those that have a sugar chain structure that is compatible with lectin LA and those that do not. As a result, at the detection site 34A, as shown in graph G21 in Figure 8, a peak is detected from protein PA that does not have a sugar chain structure compatible with lectin LA, and a peak is detected from protein PA that has a sugar chain structure compatible with lectin LA (LA affinity peak in graph G21).

[0076] Meanwhile, in the second branch channel 23, protein PB is fractionated into those that have a sugar chain structure that is compatible with lectin LB and those that do not. As a result, at the detection site 34B, as shown in graph G22 in Figure 8, peaks are detected from protein PB that does not have a sugar chain structure compatible with lectin LB and peaks from protein PB that has a sugar chain structure compatible with lectin LB (LB affinity peaks in graph G22).

[0077] In this way, the presence or absence of glycosylation mutations in multiple target substances (in this case, protein PA and protein PB) can be simultaneously detected in the first branch channel 22 and the second branch channel 23. That is, glycosylation mutations in multiple target substances can be detected in a single measurement.

[0078] In the above, the case in which the measurement chip 10 is equipped with two branched channels 22 and 23 has been described, but the measurement chip 10 may be equipped with three or more branched channels. By providing three or more branched channels, it is possible to perform multiple inspections in a single measurement by combining the above embodiments or application examples 1 to 3.

[0079] In the above embodiment, it was described that reagents S1 to S5 are pre-loaded into the flow path 20 of the measurement chip 10, but it is preferable that the reagents are filled into the measurement chip 10 by the analyzer 100.

[0080] "Analyzer According to the Second Embodiment" FIG. 9 shows a schematic configuration of an analyzer 110 according to the second embodiment. The analyzer 110 includes a reagent filling mechanism 14 in addition to an electrophoresis mechanism 12 and a control device 18. Other basic configurations of the analyzer 110 according to the second embodiment are the same as those of the analyzer 100 according to the first embodiment, so descriptions of common features will be omitted and the description will focus on differences. In FIG. 9 and subsequent figures, the same reference numerals are assigned to components equivalent to those in FIGS. 1 to 8.

[0081] The reagent filling mechanism 14 includes a dispenser 14A and a pressure applying unit 14B. In the present embodiment, a measurement chip 10A not filled with a reagent is loaded, filled with a reagent in the analyzer 110, and then supplied for analysis. In the analyzer 110, the measurement chip 10A is placed on a support base 11, and the analyzer 110 includes a movement mechanism (not shown) that moves the measurement chip 10A from a reagent filling position by the reagent filling mechanism 14 to an electrophoresis position by the electrophoresis mechanism 12.

[0082] FIG. 10 shows a schematic configuration of the measurement chip 10A. In FIG. 10, a flow path 20A of the measurement chip 10A is schematically illustrated. As shown in FIG. 10, the flow path 20A of the measurement chip 10A includes a main flow path 21, branch flow paths 22 and 23, and wells 26a to 26d, as well as wells 26e to 26g for reagent filling and wells 27a to 27d for waste liquid storage. The wells 26e to 26g, 26d, and 27a to 27d are each connected via a connection path 24 １ to 24 ８ to the main flow path 21. The reagent filling wells 26e to 26g and the waste liquid storage wells 27a to 27d are arranged on opposite sides with the main flow path 21 interposed therebetween, at positions offset from each other. The connection path 24 １ to 24 ４ are connected from the upstream side to the downstream side in the order of the connection path 24 １ , the connection path 24 ２ , the connection path 24 ３ and the connection path 24 ４ . The connection path 24 ４ corresponds to the connection path 24 of the measurement chip 10. Further, the connection path 24 ５ to 24 ８ are the connection path 24 １ to 24４ On the opposite side of the main channel 21, from the upstream side to the downstream side, there is a connecting channel 24 ５ , connecting path 24 ６ , connecting path 24 ７ and connecting path 24 ８ They are connected in this order. More specifically, the connecting path 24 ５ is the connecting path 24 １ It is connected to the main channel 21 upstream of it. Connecting channel 24 ６ is the connecting path 24 １ It is downstream of the connecting passage 24 ２ It is connected upstream of the connection point 24. ７ is the connecting path 24 ２ It is downstream of the connecting passage 24 ３ It is located further upstream. Connecting passage 24 ８ is the connecting path 24 ３ It is downstream of the connecting passage 24 ４ It is located further upstream.

[0083] The dispenser 14A dispenses reagent S1 into well 26a, reagent S2 into well 26e, reagent S3 into well 26f, reagent S4 into well 26g, and reagent S5 into wells 26b, 26c, and 26d.

[0084] The pressure application unit 14B introduces reagents S1 to S5 into the flow path 20A by applying pressure to the reagents S1 to S5 dispensed into wells 26a to 26g. The pressure application unit 14B is configured to apply pressure to reagents S1 to S5 by supplying pressurized gas while keeping wells 26a to 26g airtight. The pressure application unit 14B includes, for example, a sealing mechanism to make each well 26a to 26g airtight and an air pump to supply the gas.

[0085] As shown in Figure 11, reagents S1 to S5 are introduced into the flow path 20A by dispensing reagents S1 to S5 into wells 26a to 26g using the dispenser 14A and by pressurizing reagents S1 to S5 using the pressure application unit 14B. At this time, well 27a functions as a waste liquid reservoir for storing excess reagent S1 introduced from well 26a and excess reagent S2 introduced from well 26e. Well 27b functions as a waste liquid reservoir for storing excess reagent S2 introduced from well 26e and excess reagent S3 introduced from well 26f. Well 27c functions as a waste liquid reservoir for storing excess reagent S3 introduced from well 26f and excess reagent S4 introduced from well 26g. Well 27d functions as a waste liquid reservoir for storing excess reagent S4 introduced from well 26g, and excess reagent S5 introduced from wells 26b, 26c, and 26d.

[0086] Furthermore, in cases where reagent S5 containing different components is introduced into the first branch channel 22 and the second branch channel 23, as in Application Examples 1 to 3 of the first embodiment, the reagent filling mechanism 14 should be configured to introduce reagent S5 containing different components into the first branch channel 22 and the second branch channel 23.

[0087] For example, in Application Example 1, the reagent filling mechanism 14 introduces reagent S5 containing lectin L from well 26b, which constitutes the end of the first branch channel 22, and introduces reagent S5 without lectin L into wells 26c and 26d. In Application Example 2, the reagent filling mechanism 14 introduces reagent S5 containing lectin L1 from well 26b, which constitutes the end of the first branch channel 22, introduces reagent S5 containing lectin L2 from well 26c, which constitutes the end of the second branch channel 23, and introduces reagent S5 without lectins L1 and L2 from well 26d. Lectins L1 and L2 are examples of "different molecules that specifically bind to different structures of the substance to be tested" in the technology of this disclosure. Furthermore, in the case of application example 3, the reagent filling mechanism 14 introduces reagent S5 containing lectin LA from well 26b which constitutes the channel end of the first branch channel 22, reagent S5 containing lectin LB from well 26c which constitutes the channel end of the second branch channel 23, and reagent S5 that does not contain lectin LA or LB from well 26d. Lectin LA and LB are examples of "molecules that specifically bind to different glycans for detecting glycan mutations in proteins" in the technology of this disclosure.

[0088] The analysis method in the analysis apparatus 110 of the second embodiment is the same as the analysis method of the first embodiment.

[0089] As described above, in this embodiment, the analyzer 110 has a reagent filling mechanism 14 that introduces reagents S1 to S5 into the flow path 20A of the measuring chip 10A by pressure. With this configuration, reagents S1 to S5 can be filled into the flow path 20A without any air pockets. The user only needs to load an empty measuring chip 10A into the analyzer 110, and a series of operations for analysis are completed within the analyzer 110, thus reducing the burden on the user. Furthermore, with this configuration, reagents S1 to S5, whose conductivity changes in stages, can be arranged sequentially from upstream to downstream in the measuring chip 10A as shown in Figure 11, enabling highly accurate isokinetic electrophoresis.

[0090] "Analytical Apparatus of the Third Embodiment" Figure 12 is a schematic diagram showing the general configuration of an analytical apparatus 120 of the third embodiment according to the technology of this disclosure. This embodiment is one form of the analytical apparatus of the second embodiment. The analytical apparatus 120 of the third embodiment differs from the first embodiment in its configuration for detecting the substance to be tested from the detection site. Since the other basic configurations of the analytical apparatus 120 of the third embodiment are the same as those of the analytical apparatus 100 of the first embodiment, the explanation of the common points will be omitted and the explanation will focus on the differences.

[0091] The analyzer 100 of the first embodiment is equipped with separate detection units 16A and 16B for detection points 34A and 34B of the two branched channels 22 and 23. In contrast, the analyzer 120 of the third embodiment is equipped with only one detection unit 16. The detection unit 16 is, for example, a photodetector, and like the detection units 16A and 16B, it is equipped with, for example, an objective lens, an excitation light source, and a photodetector, and detects an optical signal as a signal. It also detects fluorescence as an optical signal, similar to the first embodiment. On the other hand, since one detection unit 16 performs detection at the two detection points 34A and 34B, the analyzer 120 of this embodiment is equipped with a moving mechanism 17 that moves the measurement chip 10 and the detection unit 16 relative to each other.

[0092] In this embodiment, the moving mechanism 17 moves the detection unit 16 between each detection point 34A, 34B in the multiple branched channels 22, 23 (within the range indicated by the double arrows). For example, the moving mechanism 17 can be a piezo stage, a piezo motor stage, or a linear stage. The moving mechanism 17 may also be configured by combining a stepping motor or a servo motor with a ball screw or a linear guide.

[0093] In the analyzer 120, the electrophoresis mechanism 12 branches the substance A to be tested, which has moved from the upstream side of the main channel 21 and reached the branching point of the branching channels 22 and 23, into the branching channels 22 and 23, and shifts the time it takes for the substance A to be tested, which has been branched into the branching channels 22 and 23, to reach the detection sites 34A and 34B in each branching channel 22 and 23. Here, "shifting the time" means controlling the time so that it is different. That is, the electrophoresis mechanism 12 controls the voltage or current applied to the channel 20 so that the time it takes for the substance A to be tested to reach the detection site 34A by gel electrophoresis in the first branching channel 22 is different from the time it takes for the substance A to be tested to reach the detection site 34B by gel electrophoresis in the second branching channel 23. In this embodiment, the electrophoresis mechanism 12 controls the voltage or current applied to the channel 20 such that the time at which the substance A to be tested reaches the detection site 34A in the first branch channel 22 is earlier than the time at which the substance A to be tested reaches the detection site 34B in the second branch channel 23.

[0094] The movement mechanism 17 moves the detection unit 16 from a position facing detection site 34A to a position facing detection site 34B between the time the substance A to be tested reaches detection site 34A in the first branch channel 22 and the time the substance A to be tested reaches detection site 34B in the second branch channel 23. As a result, one detection unit 16 detects fluorescence corresponding to the substance A to be tested from each of the detection sites 34A and 34B in the multiple branch channels 22 and 23.

[0095] As previously described, the electrophoresis mechanism 12 includes a power supply and a control unit. Here, the control method by the control unit differs from that of the first embodiment. A method for shifting the time it takes for each of the substances to be tested A, which are branched into branched channels 22 and 23, to reach the detection sites 34A and 34B will be described.

[0096] The electrophoresis mechanism 12 sequentially applies a first current to each of the multiple branch channels 22, 23, which has a relatively larger absolute value than the other branch channels where detection is not yet complete, and applies a voltage or current to the channel 20 such that the substance A to be tested in the other branch channels where detection is not yet complete does not flow back to the main channel 21. For example, the first current is first applied to the first branch channel 22 to cause the substance A to be tested to reach the detection site 34A, and then the first current is applied to the second branch channel 23. This delays the time at which the substance A to be tested reaches the detection site 34B in the second branch channel 23 compared to the time at which it reaches the detection site 34A in the first branch channel 22. Furthermore, it is preferable that the electrophoresis mechanism 12 applies voltage or current to the channel 20 such that while a first current, which has a relatively larger absolute value than the other branch channels where detection is not yet complete, flows through one branch channel, a second current, which has a relatively smaller absolute value than the first current, flows through the other branch channels where detection is not yet complete.

[0097] A specific example will be explained with reference to Figures 13 and 14, along with the steps of the analysis method.

[0098] In this embodiment, the analysis method is the same as in the first embodiment up to the formation of the concentrated layer F in the concentrated region 30. That is, steps 1 to 3 shown in Figure 4 and step 4 shown in Figure 5 are carried out similarly, so the details are omitted. As shown in step 4 in Figure 5, after the concentrated layer F is branched into a plurality of branched channels 22 and 23, in step 15 in Figure 13, the electrophoresis mechanism 12 of this embodiment first applies a first current, which has a relatively larger absolute value than the second branched channel 23, to the first branched channel 22, and applies a voltage to the channel 20 such that the substance to be tested A in the second branched channel 23 does not flow back into the main channel 21. Note that in Figures 13 and 14, the substance to be tested A is contained in the immunocomplex E. Here, the substance to be tested A is bound to the first labeled antibody D which has a negative charge, and this state is referred to as "the substance to be tested A is negatively charged." Because the substance to be tested A is negatively charged, it electrophores toward the anode. Therefore, during electrophoresis, the upstream side is the cathode and the downstream side is the anode. As a result, the current flows from downstream to upstream, in the opposite direction to the electrophoretic direction of the substance A being tested. In Figure 13 and subsequent figures, the direction of the arrows is aligned with the direction of electron flow, which is the same as the flow direction of the substance A being tested. The current values ​​are expressed with positive for flow from the anode to the cathode and negative for the opposite direction.

[0099] Here, "applying a voltage to the channel 20 such that a first current with a relatively larger absolute value than that of the second branch channel 23 flows through the first branch channel 22, and the substance A to be tested in the second branch channel 23 does not flow back into the main channel 21" does not necessarily mean applying a voltage to the entire channel 20, but rather means applying a voltage to a part or part of the channel 20 such that the condition (hereinafter referred to as condition 1) is satisfied, namely, "a first current with a relatively larger absolute value than that of the second branch channel 23 flows through the first branch channel 22, and the substance A to be tested in the second branch channel 23 does not flow back into the main channel 21."

[0100] Specifically, the electrophoresis mechanism 12 is the channel end of the main channel 21, and the well 26a that constitutes the upstream channel end of the channel 20, and the connecting passage 24 that is the downstream side of the main channel 21. ４By controlling the voltage applied to the electrodes inserted in each of the wells 26d, which constitute the end of the flow path, well 26b, which constitutes the downstream end of the first branch flow path 22, and well 26c, which constitutes the downstream end of the second branch flow path 23, a voltage that satisfies condition 1 is applied to the flow path 20.

[0101] More specifically, as shown in Figure 13 as an example, the electrophoresis mechanism 12 applies voltages of 0V to well 26d, 1010V to well 26b, and 110V to well 26c. In addition, a voltage of 10V, equal to the potential of the connection point 33 where the connecting path 24 connects to the main flow path 21, is applied to well 26a. Note that "applying a voltage of aV (where a is any number) to the well" means setting the potential of the well to aV. As a result, a voltage of 1000V is applied between both ends of the first branched flow path 22 and a voltage of 100V is applied between both ends of the second branched flow path 23. That is, the electrophoresis mechanism 12 flows a current with a relatively large absolute value through the first branched flow path 22 while flowing a current with a relatively small absolute value through the second branched flow path 23. Here, the voltage across both ends of the first branch channel 22 is 10 times the voltage across both ends of the second branch channel 23, and as a result, the first current flowing in the first branch channel 22 is approximately 10 times that of the second current flowing in the second branch channel 23. This satisfies condition 1, and the substance A to be tested undergoes gel electrophoresis in the first branch channel 22 at approximately 10 times the speed of the substance A to be tested in the second branch channel 23.

[0102] As a result, as shown in Step 16 of Figure 13, the substance A to be tested in the first branch channel 22 reaches the detection site 34A faster than the substance A to be tested in the second branch channel 23 reaches the detection site 34B. The detection unit 16 is positioned in advance facing the detection site 34A of the first branch channel 22 by a moving mechanism 17 (see Figure 12), which is not shown in Figure 13. The substance A to be tested that reaches the detection site 34A is detected by the detection unit 16. In detail, the detection unit 16 irradiates the detection site 34A with excitation light and detects the fluorescence generated from the detection site 34A by the irradiation of the excitation light.

[0103] Next, the electrophoresis mechanism 12 applies a voltage to the channel 20 that satisfies condition 2, which is "a current equal to or greater than that of the first branch channel 22 flows through the second branch channel 23," from condition 1. Simultaneously with, or before or after, the electrophoresis mechanism 12 switches from the voltage of condition 1 to the voltage of condition 2, the moving mechanism 17 (see Figure 12) moves the detection unit 16 from the detection area 34A of the first branch channel 22 to a position facing the detection area 34B of the second branch channel 23.

[0104] Here, in order to promote gel electrophoresis in the second branch channel 23, the electrophoresis mechanism 12 applies voltages of 0V to well 26d, 110V to well 26b, and 1010V to well 26c, for example, as shown in Step 17 of Figure 14. In addition, a voltage of 10V, equal to the potential of the connection point 33 where the connecting path 24 connects to the main channel 21, is applied to well 26a. As a result, a voltage of 100V is applied between both ends of the first branch channel 22 and a voltage of 1000V is applied between both ends of the second branch channel 23. The voltage between both ends of the second branch channel 23 is 10 times the voltage between both ends of the first branch channel 22, and as a result, the first current flowing through the second branch channel 23 is approximately 10 times the second current flowing through the first branch channel 22. This satisfies condition 2, and in the second branch channel 23, the substance A to be tested undergoes gel electrophoresis at approximately 10 times the speed of the substance A to be tested in the first branch channel 22.

[0105] As a result, as shown in Step 18 of Figure 14, gel electrophoresis of the substance A to be tested in the second branch channel 23 is accelerated. The substance A to be tested that reaches the detection site 34B is detected by the detection unit 16. Here again, the detection unit 16 irradiates the detection site 34B with excitation light and detects the fluorescence generated from the detection site 34B by the irradiation of the excitation light.

[0106] In the above description, the electrophoresis mechanism 12 applied a specific voltage to each of the wells 26a, 26b, 26c, and 26d in order to apply a voltage or current to the channel 20 that satisfies condition 1 or condition 2. However, the electrophoresis mechanism 12 is not limited to this method of applying a voltage to the channel 20 that satisfies condition 1 or condition 2. For example, as shown in Figure 13, the electrophoresis mechanism 12 has a connecting path 24 leading from well 26d to the main channel 21. ４ For example, a current of 22 μA (microamperes) may be applied, a first current of 20 μA may be applied to the first branch channel 22, a second current of 2 μA may be applied to the second branch channel 23, and control may be performed to ensure that the current in the main channel 21 is 0 μA (i.e., no current flows) or to electrically disconnect the main channel 21 by withdrawing the electrode from the well 26a. In this case, the electrophoresis mechanism 12 has a connection path 24 ４ The voltages applied to wells 26a, 26b, 26c, and 26d are controlled so that the above current flows through the main flow path 21, the first branch flow path 22, and the second branch flow path 23, respectively.

[0107] In Step 17 of the above analysis method, the second branch channel 23, where detection is performed second, is controlled to have a larger current flowing through it than the first branch channel 22. That is, a first current with a relatively larger absolute value than that of the first branch channel 22 flows through the second branch channel 23, and a voltage is applied to the channel 20 so that the substance A to be tested in the first branch channel 22 does not flow back into the main channel 21. However, since the inspection in the first branch channel 22 is complete, the voltage (or current) applied to the first branch channel 22 is arbitrary and not particularly restricted. For example, the same current as the second branch channel 23 may flow through the first branch channel 22, or a voltage may be applied to the well 26b so that no current flows through the first branch channel 22. Also, after the first branch channel 22 is completed, even if the voltage of the second branch channel 23 is not changed, gel electrophoresis is occurring in the second branch channel 23, albeit at a slow speed, so the voltage may not be switched, and the system may wait for the substance A to be tested to pass through the inspection site 34B.

[0108] As described above, in the analyzer 120 of this embodiment, the electrophoresis mechanism 12 shifts the time it takes for the substance to be tested A to reach the detection sites 34A and 34B in the multiple branched channels 22 and 23. As a result, a single detection unit 16 can sequentially detect signals corresponding to the substance to be tested A from each of the detection sites 34A and 34B.

[0109] Since only one detection unit 16 is needed for each of the multiple branch channels 22 and 23, i.e., each of the multiple detection points 34A and 34B, the overall configuration can be made more compact compared to the case where each of the multiple branch channels 22 and 23 has its own individual detection unit 16. Also, having only one detection unit 16 improves the flexibility of placement compared to having multiple units. Furthermore, if the detection unit 16 is a photodetector, the objective lens is larger in size than the microchannel, so if one is to be placed for each of the branch channels 22 and 23, the branch channels 22 and 23 must be spaced far apart, reducing the design flexibility of the branch channels 22 and 23. In contrast, having only one detection unit 16 provides greater design flexibility for the branch channels 22 and 23. In addition, using only one detection unit 16 can reduce the cost of the device.

[0110] In the above description, we explained the case where an analytical device for measuring a measurement chip 10 having two branched channels 22 and 23 is equipped with one detection unit 16. However, in an analytical device for measuring a measurement chip having three or more branched channels, it is not limited to having only one detection unit, but may be equipped with fewer detection units than the number of branched channels. For example, an analytical device for measuring a measurement chip having four branched channels may be equipped with two or three detection units. In this case, at least one detection unit can be moved relative to the multiple branched channels and detection can be performed from the detection point of each branched channel. This can reduce costs and allow for a more compact configuration than when a detection unit is provided for each branched channel.

[0111] In this embodiment, the electrophoresis mechanism 12 applies a voltage or current that satisfies condition 1 to the channel 20, thereby shifting the time it takes for the substance to be tested, which is branched into the branched channels 22 and 23, to reach each detection site 34A and 34B. This can be achieved by controlling the voltage applied to wells 26a, 26b, 26c, and 26d, and the time it takes for the substance to reach the detection sites 34A and 34B can be shifted without adding any additional structures.

[0112] The method for shifting the time it takes for the substance A to be tested to reach the detection sites 34A and 34B in the multiple branched channels 22 and 23 is not limited to the above embodiment. For example, the time it takes to reach the detection sites 34A and 34B may be shifted by making the distances from the branching point 29 of the multiple branched channels 22 and 23 to the detection sites 34A and 34B different. For example, in the measurement chip 10, if the length of the branched channel 22 is shorter than the length of the branched channel 23, and the distance from the branching point 29 to the detection site 34A is set to be shorter than the distance from the branching point 29 to the detection site 34B, the detection time at the detection site 34B can be delayed compared to the detection time at the detection site 34A.

[0113] Furthermore, in this embodiment, the electrophoresis mechanism 12 applies a voltage (or current) to the channel 20 such that, while a first current with a relatively larger absolute value than that of the branch channel where detection is not yet complete (here, the second branch channel 23) flows through one branch channel (here, the first branch channel 22), a second current with a relatively smaller absolute value than that of the first current flows through the branch channel where detection is not yet complete. Here, the first and second currents are currents in the same direction. By applying a relatively small second current to the branch channel where detection is not yet complete while the substance to be tested is undergoing gel electrophoresis in one branch channel through which the first current is flowing, it is possible to effectively suppress the backflow of the substance to be tested A from these branch channels into the main channel 21. In particular, when a concentrated layer F is formed in the concentrated region 30 of the main channel 21, and the concentrated layer F is branched into each of the branch channels 22 and 23, the diffusion of the concentrated layer F can be suppressed, and the decrease in the accuracy of the test in the branch channel where detection is not yet complete can be effectively suppressed.

[0114] In this embodiment, the moving mechanism 17 is a detection unit moving mechanism that moves the detection unit 16. In conventional analytical devices that detect fluorescence from a measurement chip equipped with a microchannel using a photodetector unit, a moving mechanism is sometimes provided to move the photodetector unit in order to focus the objective lens of the photodetector unit. Therefore, an existing moving mechanism can be used as a detection unit moving mechanism that moves the detection unit 16 between detection parts 34A and 34B, thereby suppressing the increase in size of the analytical device 120.

[0115] On the other hand, the moving mechanism 17 may be a measuring chip moving mechanism that moves the measuring chip 10 relative to the detection unit 16. In this case, the measuring chip moving mechanism can sequentially move each of the detection points 34A and 34B of the multiple branched channels 22 and 23 to the detection position of the detection unit 16. Furthermore, the moving mechanism 17 may comprise a detection unit moving mechanism that moves the detection unit 16 and a measuring chip moving mechanism that moves the measuring chip 10, and move the two relative to each other.

[0116] As described above, in this embodiment, the electrophoresis mechanism 12 applies a voltage (or current) to the channel 20 such that a first current, which has a relatively larger absolute value than the other branch channels (here, the second branch channel 23) through which detection is not yet complete, flows through one branch channel (here, the first branch channel 22), while a second current, which has a relatively smaller absolute value than the first current, flows through the other branch channels through which detection is not yet complete. However, it is sufficient that condition 1 is satisfied, and the embodiment is not limited to this form. Modifications 1 and 2 of the applied voltage in Step 15 are shown below.

[0117] (Modification 1) The electrophoresis mechanism 12 may be modified to Step 15A as shown in Figure 15, instead of Step 15, by applying voltages of 10V to well 26a, 0V to well 26d, 1010V to well 26b, and 10V to well 26c. This also satisfies condition 1. In this case, a voltage of 1000V is applied between both ends of the first branch channel 22, promoting gel electrophoresis of the substance to be tested A, and the substance to be tested A moves toward the detection site 34A. On the other hand, the potential at the connection point 33 is 10V, so no current flows in the second branch channel 23, and no movement of the substance to be tested A occurs within the second branch channel 23. After detection of the substance to be tested A at the detection site 34A of the first branch channel 22, voltages of 10V to well 26b and 1010V to well 26c are applied to cause gel electrophoresis of the substance to be tested A in the second branch channel 23. This allows for the detection of the target substance A at the detection site 34B of the second branch channel 23 with a time delay.

[0118] In Step 15A, instead of setting the voltage applied to wells 26a to 26d as described above, the electrophoresis mechanism 12 uses a connecting path 24 from well 26d to the main channel 21. ４ For example, a current of 20 μA may be passed through the first branch channel 22, and control may be performed so that no current flows through the second branch channel 23 and the main channel 21. In this case, the electrophoresis mechanism 12 is connected to the connection channel 24 ４ The voltages applied to wells 26a, 26b, 26c, and 26d are controlled so that the above current flows through the main flow path 21, the first branch flow path 22, and the second branch flow path 23, respectively.

[0119] In this way, even when controlling the flow so that no current flows to other branch channels 23 where detection is not yet complete while a first current for gel electrophoresis is flowing through one branch channel 22, it is possible to suppress the backflow of the substance A to be tested to the branching point in the other branch channels 23. Furthermore, if a concentrated layer F is formed in the concentrated region 30 of the main channel 21 and the concentrated layer F is branched into each of the branch channels 22 and 23, the diffusion of the concentrated layer F can be suppressed, thereby suppressing a decrease in the accuracy of the test in the other branch channels 23 where detection is not yet complete.

[0120] (Modification 2) Alternatively, the electrophoresis mechanism 12 may be configured as Step 15B as shown in Figure 16, instead of Step 15, applying voltages of 10V to well 26a, 0V to well 26d, 1010V to well 26b, and 0V to well 26c. This also satisfies condition 1. In this case, a voltage of 1000V is applied between both ends of the first branch channel 22, and for example, a current of 20.2μA flows through the first branch channel 22, promoting gel electrophoresis of the substance A to be tested, and the substance A to be tested moves toward the detection site 34A. On the other hand, the potential at the connection point 33 is 10V, and a small current flows through the second branch channel 23 from downstream to upstream (for example, 0.2μA in Figure 16). However, this is not a problem as long as the concentrated layer F (i.e., the substance A to be tested) in the second branch channel 23 does not flow back to the branch point 29.

[0121] In the case of the second modification, after detecting the substance A to be tested at the detection site 34A of the first branch channel 22, a voltage of 0V is applied to well 26b and a voltage of 1010V is applied to well 26c to perform gel electrophoresis on the substance A to be tested in the second branch channel 23. This allows for the detection of the substance A to be tested at the detection site 34B of the second branch channel 23 with a time delay.

[0122] In the third embodiment and its modifications 1 and 2, the application of voltage (or current) was described assuming that the substance A to be tested is negatively charged. Therefore, the direction of movement of substance A by electrophoresis coincides with the direction of electron flow. On the other hand, if the substance to be tested is positively charged, the direction of movement of the substance by electrophoresis will coincide with the direction of current. This can be achieved by reversing the sign of the voltage (or current) in the above description.

[0123] Furthermore, the analyzer 120 of the third embodiment may be configured to include a reagent filling mechanism 14, similar to the analyzer 110 of the second embodiment, and to be loaded with a measurement chip 10A.

[0124] In the analytical devices 100, 110, and 120 of the first to third embodiments described above, a concentrated layer F is formed in the main channel 21 of the measurement chips 10 and 10A, and then the concentrated layer F is branched into multiple branch channels 22 and 23. However, the formation of the concentrated layer F is not essential. Even with a configuration that performs gel electrophoresis from the main channel 21 without forming a concentrated layer F, the effect of being able to perform multiple tests in parallel can be obtained.

[0125] In the analyzers 100, 110, and 120 of the first to third embodiments described above, the detection units 16A, 16B, and 16 are each photodetector units, and they detect fluorescence from a fluorescent label as a signal corresponding to the substance to be tested. However, the system is not limited to this configuration. In addition to fluorescent labels, the system may be configured to detect optical signals from enzyme labels or gold colloid labels. These can be achieved by changing the label attached to the second-labeled antibody. Alternatively, the signal corresponding to the substance to be tested is not limited to an optical signal. For example, instead of a fluorescent label, a radioactive label may be used, and the system may be configured to detect radiation as a signal corresponding to the substance to be tested using a detection unit that includes a radiation detector. Furthermore, instead of a fluorescent label, a magnetic particle label may be used, and the system may be equipped with a detection unit that magnetically detects the substance to be tested.

[0126] In the analysis devices 100, 110, and 120 of the first to third embodiments described above, the measuring tips 10 and 10A used are described as having two branched channels 22 and 23, but they may also be configured to have three or more branched channels.

[0127] Here, we will briefly explain an example in which, when a single detection unit 16 detects the three branched channels sequentially using a measuring chip equipped with three branched channels, as in the analytical apparatus 120 of the third embodiment, the arrival times of the target substance A in each branched channel are made to differ from one another. For the sake of explanation, the three branched channels will be referred to as branched channel 1, branched channel 2, and branched channel 3, and detection will be performed in the order of branched channel 1, branched channel 2, and branched channel 3. An example of the current flowing through each branched channel 1 to 3 will be explained. First, in order to detect branched channel 1, a larger current is flowed through branched channel 1 than that of branched channels 2 and 3, which have not yet been detected. For example, branched channel 1: 20 μA, branched channel 2: 2 μA, branched channel 3: 2 μA. Next, in order to detect branched channel 2, a larger current is flowed through branched channel 2 than that of branched channel 3, which has not yet been detected. For example, branched channel 1: arbitrary, branched channel 2: 20 μA, branched channel 3: 2 μA. In this case, it is acceptable whether or not current is flowing through branched channel 1, which has already been detected. Next, detection is performed in branch channel 3. In this case, a current greater than 0 μA (a current that causes gel electrophoresis) is passed through branch channel 3. For example, branch channel 1: arbitrary, branch channel 2: arbitrary, branch channel 3: 0 μA or more. Since branch channel 3 is the last channel to be detected, it is sufficient that the time it takes for the target substance to reach the detection site is later than that of branch channels 1 and 2.

[0128] Furthermore, the measurement chip may be configured to branch from one branch channel into multiple branch channels.

[0129] Figure 17 shows a schematic diagram of a modified flow path 20B. Flow path 20B comprises a main flow path 21 as an electrophoretic flow path, two branched flow paths 131 and 132 branching from the main flow path 21 at the first branching point 29A, two branched flow paths 141 and 142 branching from branched flow path 131 at the second branching point 29B, and two branched flow paths 143 and 144 branching from branched flow path 132 at the third branching point 29C. One end of each branched flow path 141 to 144 is a flow path end, and wells 126a, 126b, 126c, and 126d are formed therein. Similar to wells 26a and 26d, electrodes are inserted into wells 126a to 126d as appropriate when a voltage is applied by the electrophoresis mechanism 12. In this case, the electrophoresis mechanism 12 controls the direction and magnitude of the voltage or current flowing in each region of the channel 20 by adjusting the voltage applied to the wells 26a, 26d, and 126a to 126d.

[0130] In channel 20B, a concentration region 30 is provided from the upstream side of the main channel 21 to the branching point 29. Then, four branch channels 141 to 144, each having a channel end, constitute a component separation region 32, each having detection sites 145A to 145D. In this way, when the electrophoresis channel has multiple stepwise branching points 29A to 29C, the branch channels 141 to 144, each having a downstream channel end, become the component separation region 32. Therefore, when such a channel 20B is provided, the electrophoresis mechanism 12 controls the applied voltage to wells 26a, 26d, and 126 to 126d so that after the concentration layer F is formed in the concentration region 30, the concentration layer F branches at the first branching point 29A, and then branches again at the second branching point 29B and the third branching point 29C, a voltage or current for gel electrophoresis is applied to each branch channel 141 to 144.

[0131] As an analytical device for measuring a measuring chip having a flow path 20B, a detection unit may be provided at each of the detection points 145A to 145D of the plurality of branched flow paths 141 to 144, as in the analytical device 100 of the first embodiment. In this case, signals corresponding to the substance A to be tested can be detected in parallel from all detection points 145A to 145D.

[0132] Furthermore, the analytical apparatus for measuring the measuring chip having the flow path 20B may be equipped with only one detection unit 16, as in the analytical apparatus 120 of the third embodiment. In this case, the electrophoresis mechanism 12 controls the time it takes for the substance to be tested A to reach each of the detection sites 145A to 145D of the multiple branched flow paths 141 to 144 to be different from each other. For example, the voltage applied to the wells 26a, 26d, 126a to 126d is controlled so that the times t1, t2, t3, and t4 at which the substance to be tested A, which branched off at the first branching point 29A, reaches each of the detection sites 145A to 145D are, for example, t1 < t2 < t3 < t4. First, a first current with a relatively large absolute value is passed through branch channel 141, while a second current with a relatively small absolute value is passed through the other branch channels 142, 143, and 144. Here, branch channels 142, 143, and 144 correspond to branch channels where detection has not yet been completed. This promotes gel electrophoresis in branch channel 141, allowing the target substance A to be detected at detection site 145A. Next, a first current with a relatively larger absolute value than the other branch channels 143 and 144 where detection has not yet been completed is passed through branch channel 142, while a second current with a relatively small absolute value is passed through branch channels 143 and 144. This promotes gel electrophoresis in branch channel 142, allowing the target substance A to be detected at detection site 145B. Next, a first current with a relatively larger absolute value than the other branch channels 144 where detection has not yet been completed is passed through branch channel 143, while a second current with a relatively small absolute value is passed through branch channel 144. This promotes gel electrophoresis in the branch channel 143, allowing the target substance A to be detected at the detection site 145C. Next, a larger current than the one previously flowing is applied to the branch channel 144, promoting gel electrophoresis in the branch channel 144, and allowing the target substance A to be detected at the detection site 145D. For the branch channel 144, which is the last to be detected, it is not necessarily required to apply a relatively larger current than the other branch channels; it is sufficient to apply a current that causes the target substance A to flow toward the detection site, even at a slower speed.

[0133] Furthermore, in each of the above embodiments, for example, the power supply control by the control unit of the electrophoresis mechanism 12, the movement mechanism 17, or the control device 18 of the detection units 16, 16A, 16B include a processor. The processor can be any type of processor having the following various hardware structures.

[0134] The various types of processors mentioned above include, as previously stated, CPUs, which are general-purpose processors that execute software (programs) and function as various processing units, as well as programmable logic devices (PLDs), such as FPGAs (Field Programmable Gate Arrays), which are processors whose circuit configuration can be changed after manufacturing, and dedicated electrical circuits, such as ASICs (Application Specific Integrated Circuits), which are processors with circuit configurations specifically designed to perform particular processing.

[0135] A single processing unit may be composed of one of these various processors, or it may be composed of a combination of two or more processors of the same or different types (for example, a combination of multiple FPGAs, or a combination of a CPU and an FPGA). Alternatively, multiple processing units may be composed of a single processor.

[0136] Examples of configuring multiple processing units with a single processor include, firstly, a configuration where one or more CPUs and software are combined to form a single processor, as exemplified by client and server computers, and this processor functions as multiple processing units. Secondly, a configuration using a processor that realizes the functions of the entire system, including multiple processing units, on a single IC (Integrated Circuit) chip, as exemplified by System-on-a-Chip (SoC). Thus, various processing units are configured, in terms of hardware structure, using one or more of the above-mentioned various processors.

[0137] Furthermore, the hardware structure of these various processors can more specifically utilize electrical circuits, which are combinations of circuit elements such as semiconductor devices.

[0138] Furthermore, although the above embodiments describe a configuration in which the control program 85 is pre-stored (installed) in the storage unit 84, the invention is not limited to this configuration. The control program 85 may be provided in the form of a recording medium such as a CD-ROM (Compact Disc Read Only Memory), DVD-ROM (Digital Versatile Disc Read Only Memory), or USB (Universal Serial Bus) memory. Alternatively, the control program 85 may be provided in the form of a download from an external device via a network.

[0139] The technology of this disclosure is not limited to the embodiments described above, and various configurations can be adopted as long as they do not deviate from the gist of the technology of this disclosure. Furthermore, the technology of this disclosure extends not only to the program for executing the analytical method in the analytical instrument, but also to a computer-readable storage medium for non-temporarily storing the program.

[0140] The descriptions and illustrations presented above are detailed explanations of the technical aspects of this disclosure and are merely examples of the technical aspects. For example, the above descriptions of the structure, function, operation, and effect are examples of the structure, function, operation, and effect of the technical aspects of this disclosure. Therefore, it goes without saying that you may delete unnecessary parts, add new elements, or replace elements in the descriptions and illustrations presented above, as long as you do not deviate from the essence of the technical aspects of this disclosure. Furthermore, in order to avoid confusion and facilitate understanding of the technical aspects of this disclosure, explanations of common technical knowledge and the like that do not require special explanation to enable the implementation of the technical aspects of this disclosure have been omitted from the descriptions and illustrations presented above.

[0141] The disclosure of Japanese Patent Application No. 2025-057162, filed on 28 March 2025, is incorporated herein by reference in its entirety. All documents, patent applications and technical standards described herein are incorporated herein by reference to the same extent as if each individual document, patent application and technical standard were specifically and individually noted to be incorporated by reference.

[0142] The above explanation allows us to understand the technologies related to the following first and second supplementary items.

[0143] [First Appendix 1] An analytical apparatus for detecting a target substance in a sample while performing component separation by electrophoresis on a sample supplied to the flow channel of a measuring chip equipped with a flow channel, wherein the flow channel of the measuring chip includes a main flow channel and a plurality of branched flow channels branching off from the main flow channel, and comprises an electrophoresis mechanism for electrophoresing the target substance and a detection unit for detecting the target substance, wherein the electrophoresis mechanism forms a concentrated layer in the main flow channel by concentrating the target substance, branches the concentrated layer into a plurality of branched flow channels, performs gel electrophoresis on the target substance in the branched flow channels, and the detection unit detects the target substance from each of the plurality of branched flow channels. [First Appendix 2] The analytical apparatus according to First Appendix 1, wherein the electrophoresis mechanism forms a concentrated layer by performing constant-rate electrophoresis on the target substance in the main flow channel. [1st Appendix 3] The electrophoresis mechanism has a detection function to detect the position of the concentrated layer in the main channel, and initiates gel electrophoresis in the branched channel by switching the channel end to which voltage or current is applied among a plurality of channel ends of the channel according to the position of the concentrated layer. [1st Appendix 4] The analyzer according to any one of the 1st Appendix 1 to 1st Appendix 3, having a reagent filling mechanism for introducing reagents containing different components into at least two of the branched channels of a plurality of branched channels. [1st Appendix 5] The analyzer according to the 1st Appendix 4, having a reagent filling mechanism for introducing reagents into the branched channels by dispensing reagents into the ends of the branched channels and applying pressure. [1st Appendix 6] The analyzer according to the 1st Appendix 4 or 1st Appendix 5, comprising different molecules that specifically bind to different structures of the substance to be tested, as different components. [1st Appendix 7] The analytical apparatus according to 1st Appendix 6, wherein the substance to be tested is a specific protein, and the molecules are different molecules that specifically bind to different sugar chains for detecting sugar chain mutations in the protein. [1st Appendix 8] The analytical apparatus according to any one of 1st Appendix 1 to 1st Appendix 7, wherein the detection unit is a photodetector that detects an optical signal corresponding to the substance to be tested. [1st Appendix 9] The analytical apparatus according to 1st Appendix 8, wherein the optical signal is fluorescence.

[0144] [Second Appendix 1] An analytical apparatus for detecting a target substance in a sample while performing component separation by electrophoresis on a sample supplied to the flow channel of a measuring chip equipped with a flow channel, wherein the flow channel of the measuring chip includes a main flow channel and a plurality of branched flow channels branching off from the main flow channel, and comprises an electrophoresis mechanism for electrophoresing the target substance in the main flow channel and the plurality of branched flow channels, and a single photodetector unit for detecting the target substance, wherein the electrophoresis mechanism branches the target substance from the main flow channel to the plurality of branched flow channels, shifts the time it takes for the target substance to reach the detection site in each of the plurality of branched flow channels, and the single detection unit sequentially detects the target substance from the detection site in each of the plurality of branched flow channels. [Second Appendix 2] The electrophoresis mechanism flows a first current, which has a relatively larger absolute value than other branch channels in which detection has not been completed, sequentially through each of the multiple branch channels, and applies a voltage or current to the channels that prevents the substance to be tested from flowing back into the main channel from other branch channels in which detection has not been completed, thereby shifting the time it takes for the substance to be tested branched into the branch channels to reach the detection site in each branch channel, as described in Second Appendix 1. [Second Appendix 3] The electrophoresis mechanism applies a voltage or current to the channels that allows a second current, which has a relatively smaller absolute value than the first current, to flow in the same direction as the first current, while the first current is flowing through one branch channel, as described in Second Appendix 2. [Second Appendix 4] The electrophoresis mechanism is equipped with a detection unit movement mechanism that moves one detection unit between each detection site in the multiple branch channels, as described in any one of Second Appendix 1 to Second Appendix 3. [Second Appendix 5] An analytical apparatus according to any one of Second Appendix 1 to 4, comprising a measurement chip moving mechanism that moves the measurement chip so that each detection site of a plurality of branched channels is sequentially positioned at a detection position by a single photodetector. [Second Appendix 6] An analytical apparatus according to any one of Second Appendix 1 to 5, wherein the detection unit is a photodetector that detects an optical signal corresponding to the substance to be tested. [Second Appendix 7] An analytical apparatus according to Second Appendix 6, wherein the optical signal is fluorescence.[Second Appendix 8] The electrophoresis mechanism forms a concentrated layer in the main channel by concentrating the substance to be tested, branches the concentrated layer into multiple branch channels, and then performs gel electrophoresis on the substance to be tested in the branch channels, as described in any one of the Second Appendix 1 to 7. [Second Appendix 9] The electrophoresis mechanism forms a concentrated layer by performing constant-velocity electrophoresis on the substance to be tested in the main channel, as described in the Second Appendix 8. [Second Appendix 10] The electrophoresis mechanism has a detection function to detect the position of the concentrated layer in the main channel, and starts gel electrophoresis in the branch channels by switching the channel end to which voltage or current is applied among multiple channel ends of the channel according to the position of the concentrated layer.

Claims

1. An analytical apparatus for detecting a target substance in a sample while performing component separation by electrophoresis on a sample supplied to the channel of a measuring chip equipped with a channel, wherein the channel of the measuring chip includes a main channel and a plurality of branch channels branching from the main channel, and comprises an electrophoresis mechanism for electrophoresing the target substance, and a detection unit for detecting the target substance, wherein the electrophoresis mechanism forms a concentrated layer in the main channel in which the target substance is concentrated, branches the concentrated layer into the plurality of branch channels, performs gel electrophoresis on the target substance in the branch channels, and the detection unit detects the target substance from each of the plurality of branch channels.

2. The electrophoresis mechanism forms the concentrated layer by performing constant electrophoresis on the substance to be tested in the main channel, according to claim 1.

3. The electrophoresis mechanism has a detection function for detecting the position of the concentrated layer in the main channel, and initiates gel electrophoresis in the branched channel by switching the channel end to which voltage or current is applied among a plurality of channel ends of the channel according to the position of the concentrated layer.

4. The analytical apparatus according to claim 1, further comprising a reagent filling mechanism for introducing reagents containing different components into at least two of the plurality of branched channels.

5. The analytical apparatus according to claim 4, wherein the reagent filling mechanism dispenses the reagent into the branched channel by dispensing the reagent into the end of the branched channel and applying pressure.

6. The analytical apparatus according to claim 4, wherein the aforementioned distinct components include distinct molecules that specifically bind to distinct structures of the substance to be tested.

7. The analytical apparatus according to claim 6, wherein the substance to be tested is a specific protein, and the different molecules are molecules that specifically bind to different sugar chains for detecting sugar chain mutations of the protein.

8. The analytical apparatus according to any one of claims 1 to 7, wherein the detection unit is a photodetector that detects an optical signal corresponding to the substance to be tested.

9. The analytical apparatus according to claim 8, wherein the optical signal is fluorescence.

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