Probe

Abstract

A probe attached to a flow cell having a first flow path through which a fluid containing a substance to be measured for physical property data of a spectroscopic analysis device flows, the probe being disposed in a state in which a tip portion protrudes from the first flow path, the tip portion having a configuration in which the tip portion protrudes from the first flow path when viewed from the flow direction of the fluid. The area of the flow path through which the fluid flows at the protruding portion of the tip portion is equal to or greater than the area of the flow path at the narrowest portion where the area as viewed from the flow direction of the fluid becomes narrowest among the flow paths through which the fluid flows downstream of the tip portion.

Description

Technical Field

[0001] The technology of this invention relates to a probe. Background Technology

[0002] The spectrophotometer includes a probe. An optical system is built into the tip of the probe for irradiating the substance being measured with measurement light and capturing the reflected light from the substance. As described in Japanese Patent Application Publication No. 2020-511635, the probe (marked as an optical analysis device connector in Japanese Patent Application Publication No. 2020-511635) is mounted in a flow cell. Japanese Patent Application Publication No. 2020-511635 discloses a method in which a spherical lens, as part of the optical system disposed at the tip of the probe, protrudes into the flow path of the flow cell through which the fluid containing the substance being measured flows. Summary of the Invention

[0003] The technical problem to be solved by the invention As described in Japanese Patent Application Publication No. 2020-511635, when the tip of the probe protrudes into the flow path of the flow cell, this protruding portion becomes a constriction where the area of ​​the flow path, viewed from the direction of fluid flow, is narrower than other portions. Because this constriction generates eddies, it may damage the substance being measured within the fluid. However, Japanese Patent Application Publication No. 2020-511635 does not provide countermeasures to control the damage to the substance being measured caused by the protruding portion within acceptable limits.

[0004] One embodiment of the technology involved in this invention provides a probe capable of controlling damage to the substance being measured within an acceptable range.

[0005] means for solving technical problems The probe of the present invention is installed in a flow cell having a first flow path through which fluid flows, the fluid containing the substance for which the physical property data of the spectrophotometer is to be measured, and the probe is configured such that its front end protrudes from the first flow path, wherein the front end has a structure such that, when viewed from the direction of fluid flow, the area of ​​the flow path through the protruding part of the front end is greater than or equal to the area of ​​the narrowest part of the flow path through which the fluid flows downstream of the front end, when viewed from the direction of fluid flow.

[0006] Preferably, the flow cell is connected to an inflow path that allows fluid to flow from the outside into the first flow path and an outflow path that allows fluid from the first flow path to flow out to the outside, with the narrowest part existing in the outflow path.

[0007] Preferably, a second flow path through which fluid flows is formed at the front end.

[0008] Preferably, the fluid flow direction in the second flow path is the same as the fluid flow direction in the first flow path.

[0009] The second flow path preferably has an inlet and an outlet for the fluid that are surrounded by walls.

[0010] Raman spectral data are preferred for physical property data.

[0011] The fluid is preferably any one of cell culture medium, culture supernatant, purification solution and culture medium.

[0012] Invention Effects According to the technology of the present invention, it is possible to provide a probe that can control the damage to the substance being measured within an acceptable range. Attached Figure Description

[0013] Figure 1 This is a graph showing the Raman spectral data of antibodies and other substances in the culture supernatant obtained in the cell culture section, measured by a measurement system.

[0014] Figure 2 This is a diagram representing the excitation light and the Raman scattered light.

[0015] Figure 3 It is an exploded three-dimensional view of the flow cell, the spectrophotometer probe unit, and the measuring head.

[0016] Figure 4 This is a dissected view of the flow cell, the spectrophotometer probe unit, and the measuring head.

[0017] Figure 5 It is a cross-sectional view of the flow cell, the probe unit for spectrophotometry, and the measuring head.

[0018] Figure 6 This is a diagram showing the optical system built into the measuring head.

[0019] Figure 7 This is an explanatory diagram showing the size relationship of the flow path areas of each part of the flow pool and the narrowest part of the second conveyor path. Detailed Implementation

[0020] As an example, such as Figure 1 As shown, the assay system 2 includes a flow cell 10 and a Raman spectrometer 11. The assay system 2 is, for example, assembled in the cell culture section 12 of a pharmaceutical manufacturing system for active pharmaceutical ingredients. The cell culture section 12 has a culture tank 13 and a decellularized filter 14. Cell culture medium 15 is stored in the culture tank 13. The Raman spectrometer 11 is an example of a "spectral analysis apparatus" according to the technology of this invention.

[0021] Antibody-producing cells 16 are seeded in culture tank 13 and cultured in cell culture medium 15. Antibody-producing cells 16 are, for example, cells established by integrating an antibody gene into a host cell such as Chinese hamster ovary cells (CHO cells). During culture, antibody-producing cells 16 produce immunoglobulins, i.e., antibodies 17. Therefore, antibody-producing cells 16 and antibodies 17 are present in cell culture medium 15. Antibody 17 is, for example, a monoclonal antibody, which becomes an effective component of a biopharmaceutical. Antibody 17 is an example of the "analyte substance" involved in the technology of this invention.

[0022] The first transport path 18 is connected to the culture tank 13. A cell removal filter 14 is disposed in the first transport path 18. The cell removal filter 14, for example, uses a tangential flow filtration (TFF) method to capture antibody-producing cells 16 in the cell culture medium 15 with a filter membrane (not shown) and removes the antibody-producing cells 16 from the cell culture medium 15. Furthermore, the cell removal filter 14 allows antibodies 17 to pass through. Therefore, downstream of the cell removal filter 14 in the first transport path 18, the flow is primarily of cell culture medium 15 containing antibodies 17. The cell culture medium 15 obtained by removing antibody-producing cells 16 through the cell removal filter 14 is referred to as culture supernatant. Hereinafter, the cell culture medium 15 obtained by removing antibody-producing cells 16 through the cell removal filter 14 will be labeled as culture supernatant 15A. Culture supernatant 15A is an example of a "fluid" according to the technology of this invention. In addition to antibody 17, the culture supernatant 15A also contains cell-derived proteins, cell-derived DNA (deoxyribonucleic acid), antibody 17 agglomerates, or viruses. These cell-derived proteins, cell-derived DNA, antibody 17 agglomerates, and viruses are also examples of the "analyte material" involved in the technology of this invention. Furthermore, the decellularization filter 14 can also be an alternating tangential flow filtration (ATF) filter.

[0023] The flow-through cell 10 is connected to the first delivery path 18 downstream of the decellularized filter 14. Culture supernatant 15A flows into the flow-through cell 10 from the first delivery path 18 at a predetermined flow rate. The first delivery path 18 is an example of the "flow path" and "inflow path" involved in the technology of this invention. A delivery pump (not shown) is provided downstream of the decellularized filter 14 in the first delivery path 18 (between the decellularized filter 14 and the flow-through cell 10). The delivery pump delivers culture supernatant 15A to the flow-through cell 10 at a flow rate of 200 cc / min or more, for example, 300 cc / min.

[0024] The second transport path 19 is also connected to the flow cell 10. The culture supernatant 15A flowing into the flow cell 10 from the first transport path 18 flows out to the second transport path 19. The second transport path 19 is an example of the "flow path" and "outflow path" involved in the technology of this invention. The second transport path 19 is, for example, connected to a purification unit that uses a chromatographic apparatus to purify antibody 17 from culture supernatant 15A, and transports the culture supernatant 15A from the flow cell 10 to the purification unit.

[0025] As an example, such as Figure 2 As shown, the Raman spectrometer 11 is a device for evaluating a substance M using the characteristics of Raman scattered light RSL. When an excitation light EL is irradiated onto a substance M, Raman scattered light RSL with a different wavelength than the excitation light EL is generated through the interaction between the excitation light EL and the substance M. The wavelength difference between the excitation light EL and the Raman scattered light RSL corresponds to the energy portion of the molecular vibrations of the substance M. Therefore, Raman scattered light RSL with different wavenumbers can be obtained between substances M with different molecular structures. Furthermore, the Raman scattered light RSL preferably uses Stokes lines and anti-Stokes lines.

[0026] Return to Figure 1 The Raman spectrometer 11 consists of a measuring head 25 and an analyzer 26. The measuring head 25 is connected to the analyzer 26 via a cable 27. The measuring head 25 is mounted on the flow cell 10. In the following description, the measuring head 25 side will be referred to as "upper" or "base end", and the flow cell 10 side will be referred to as "lower", "bottom", or "front end".

[0027] The analyzer 26 has a built-in light source that emits excitation light EL. The excitation light EL emitted from the light source is guided to the measuring head 25 via cable 27. The measuring head 25 emits the excitation light EL from its front end. The excitation light EL irradiates the culture supernatant 15A flowing through the flow cell 10. Raman scattered light RSL is generated through the interaction between the excitation light EL and antibodies 17 and the like in the culture supernatant 15A. The measuring head 25 receives the Raman scattered light RSL. The Raman scattered light RSL received by the measuring head 25 is output to the analyzer 26 via cable 27.

[0028] Analyzer 26 decomposes the Raman scattered light RSL by wavenumber, deriving the intensity value of the Raman scattered light RSL for each wavenumber, thereby generating Raman spectral data 28. Raman spectral data 28 contains data recording the intensity values ​​of the Raman scattered light RSL for each wavenumber. Figure 1 In the middle, Raman spectral data 28 are at 1cm -1 Scale-derived wavenumber 700cm -1 ~1800cm -1 Data on the intensity values ​​of Raman scattered light RSL within a range. Furthermore, the graph shown at the bottom of Raman spectral data 28 is a graph plotting the intensity values ​​of Raman spectral data 28 by wavenumber and connecting them with lines. Raman spectral data 28 is an example of "physical property data" involved in the technology of this invention.

[0029] In this way, the measurement system 2 causes the culture supernatant 15A obtained from the culture tank 13 in the cultured antibody production cells 16 to flow cell 10. Then, the culture supernatant 15A flowing through the flow cell 10 is irradiated with excitation light EL by the measuring head 25, thereby measuring the Raman spectral data 28 of the antibody 17 and the like in the culture supernatant 15A.

[0030] Raman spectroscopy data 28 is used to predict the state of the analyte, such as the concentration of antibody 17 in the culture supernatant 15A. For example, a machine learning model can be used to output the concentration of antibody 17 based on the input of Raman spectroscopy data 28. Furthermore, the concentration of aggregates in the culture supernatant 15A can be predicted as a measure of the state of the analyte. It is also possible to predict density, or other parameters, in addition to concentration.

[0031] As an example, such as Figure 3 , Figure 4 and Figure 5 As shown, the flow cell 10 is a cuboid component having a straight, circularly shaped flow path 30 at its internal center. The flow path 30 is an example of the "flow path" and "first flow path" involved in the technology of this invention. The flow cell 10 is made of a metal such as Hastelloy. Alternatively, the flow cell 10 can be made of a resin such as a polyolefin resin. In the case of a resin, the flow cell 10 can be for single use.

[0032] A cylindrical boss-shaped first connecting portion 31 and a second connecting portion 32 are provided on opposite end faces of the flow cell 10. The first connecting portion 31 has an inlet 33 for the flow path 30, and the second connecting portion 32 has an outlet 34 for the flow path 30. The direction parallel to the flow path 30 from the inlet 33 to the outlet 34 is the flow direction FD1 of the culture supernatant 15A in the flow path 30. The flow direction FD1 is an example of the "fluid flow direction in the first flow path" involved in the technology of this invention.

[0033] The first transport path 18 and the first connecting portion 31 are liquid-tightly connected via a ferrule connector. The ferrule connector consists of a ferrule (not shown) formed at one end of the first transport path 18 and one end of the first connecting portion 31, a gasket (not shown) clamped in a groove of the ferrule, and a first clamping member 35 that secures one end of the first transport path 18 and one end of the first connecting portion 31. Similarly, the second transport path 19 and the second connecting portion 32 are liquid-tightly connected via a ferrule connector including a second clamping member 36. Furthermore, parallel threads or tapered threads can be used in the connection between the first transport path 18 and the first connecting portion 31 and the second transport path 19 and the second connecting portion 32.

[0034] The first delivery path 18 has the same diameter from the decellularization filter 14 to the end connected to the first connection portion 31. Furthermore, the end of the first delivery path 18 connected to the first connection portion 31 is an inverted cone shape, with its diameter gradually widening from the upstream side (cell culture section 12 side) towards the downstream side (flow chamber 10 side) of the flow direction FD1. Conversely, the end of the second delivery path 19 connected to the second connection portion 32 is a cone shape, with its diameter gradually narrowing from the upstream side (flow chamber 10 side) towards the downstream side (purification section side) of the flow direction FD1. The second delivery path 19 then maintains this narrowing diameter up to the purification section. The narrowing point in the second delivery path 19 is the narrowest point 41, where the area observed from the flow direction FD1 is narrowest in the flow path through which the culture supernatant 15A flows downstream of the tip 49 of the probe 45 (described later).

[0035] A mounting hole 37 is provided at the center of the upper surface of the flow cell 10. The mounting hole 37 is a circular hole for detachably mounting a spectrophotometer probe unit (hereinafter referred to as "unit") 38 in the flow cell 10, and a thread 39 is cut into its inner wall surface. The mounting hole 37 and the fitting hole 40 (see reference) Figure 4 and Figure 5 Connected to the mounting hole 37. The fitting hole 40 is also a circular hole, and its center coincides with that of the mounting hole 37. The diameter of the fitting hole 40 is smaller than that of the mounting hole 37. The fitting hole 40 passes through the flow path 30.

[0036] Unit 38 comprises a probe 45, a rotation limiting member 46, and a connecting member 47. Unit 38 may be made of a metal such as Hastelloy. Alternatively, unit 38 may be made of a resin such as a polyolefin resin. In the case of a resin-based unit, unit 38 may be for single use only.

[0037] The probe 45 is a cylindrical component with a diameter matching that of the fitting hole 40. The probe 45 is inserted into the mounting hole 37 and fitted into the fitting hole 40 by the operator of the measuring system 2. Thus, the probe 45 is disposed within the flow cell 10. With the rotation limiting member 46 not installed in the mounting hole 37 but only inserted into the mounting hole 37 and fitted into the fitting hole 40, the probe 45 can rotate 360° circumferentially. Here, "consistent" means not only completely consistent, but also consistent in the sense of an error that includes the error generally permissible in the technical field to which the present invention pertains and does not depart from the technical spirit of the present invention. The error referred to here is preferably ±10%, more preferably ±5%.

[0038] The probe 45 has a main body 48 and a front end 49. The main body 48 has a linear and circular optical path 50 at its internal center. Excitation light EL and Raman scattered light RSL pass through the optical path 50. Excitation light EL passes through the optical path 50 from the measuring head 25 and the main body 48 toward the front end 49. Conversely, Raman scattered light RSL passes through the optical path 50 from the front end 49 toward the main body 48 and the measuring head 25.

[0039] On the outer circumferential surface of the upper part of the main body 48, a radially recessed groove 51 is formed around the entire circumference. The front end of the bolt 53 presses against the bottom surface 52 of the groove 51 (see reference). Figure 5 ).

[0040] Furthermore, a radially protruding flange 54 is formed on the outer peripheral surface of the central portion of the main body 48. When the probe 45 is inserted into the mounting hole 37 and fitted into the fitting hole 40, the lower surface of the flange 54 and the bottom surface 55 of the mounting hole 37 (see reference) Figure 4 and Figure 5 The flange 54 acts as a stop to prevent the main body 48 of the probe 45, which is located higher than the flange 54, from being pulled out from the bottom surface 55.

[0041] A circular groove 56 is formed on the lower surface of the flange portion 54 (see reference). Figure 4 and Figure 5 An O-ring 57 is embedded in the groove 56. The O-ring 57 is made of elastic rubber. The O-ring 57 is flattened between the bottom surface 55 of the mounting hole 37 and the groove 56 to prevent the culture supernatant 15A flowing through the flow path 30 from leaking out. In addition, although not shown in the figure, an O-ring for preventing leakage of culture supernatant 15A is also provided between the main body 48 and the front end 49.

[0042] An optical system 60 is built into the lower part of the main body 48. The optical system 60 is positioned on its optical axis OA1 (see reference). Figure 4 and Figure 5 The position is consistent with the center of optical path 50.

[0043] The optical system 60 consists of a hemispherical lens 61 and a transparent plate 62. The hemispherical lens 61 is a lens that is literally hemispherical, for example, made of sapphire glass or quartz glass. The hemispherical lens 61 has a hemispherical incident surface for the excitation light EL and a planar exit surface for the excitation light EL.

[0044] The transparent plate 62 has an incident surface and an exit surface 63 for excitation light EL that are parallel to each other (see reference). Figure 4 and Figure 5 The disc is made of, for example, sapphire glass or quartz glass. Here, "parallel" means not only perfectly parallel, but also parallel in the sense of an error that is generally permissible in the technical field to which this invention pertains and does not deviate from the technical spirit of this invention. The error referred to here is preferably ±10%, more preferably ±5%.

[0045] The exit surface of the hemispherical lens 61 and the incident surface of the transparent plate 62 have the same curvature (0 in this case). The exit surface of the hemispherical lens 61 and the incident surface of the transparent plate 62 can be fixedly joined by an adhesive or the like, or they can be held at the front end 49 in a state where they are simply joined together without relying on an adhesive or the like. Here, "same" means not only completely identical, but also identical in the sense of error that is generally permissible in the technical field to which the present invention pertains and does not depart from the technical spirit of the present invention. The error referred to here is preferably ±10%, more preferably ±5%.

[0046] The front end portion 49 protrudes from the fitting hole 40 into the flow path 30. Therefore, when viewed from the upstream side of the flow path 30 of the fitting hole 40, the portion of the fitting hole 40 becomes a constriction. The portion of the fitting hole 40 is an example of the "protruding front end portion" involved in the technology of this invention.

[0047] The front end 49 is cylindrical, with its upper side connected to the main body 48 and open, and its lower side covered by a flat bottom plate 64 (see reference). Figure 4 and Figure 5 The base plate 64 is enclosed, and its surroundings are enclosed by a peripheral plate erected vertically upwards from the base plate 64. At 180° symmetrical positions in the center of the peripheral plate are inlet 65 and outlet 66 for the culture supernatant 15A. The inlet 65 and outlet 66 are rectangular, or more precisely, square, and are surrounded by walls. The space formed by these inlet 65 and outlet 66, and by the base plate 64 and peripheral plate, constitutes the flow path 67 through which the culture supernatant 15A flows (see reference). Figure 4 and Figure 5The direction parallel to the flow path 67 from the inlet 65 to the outlet 66 is the flow direction FD2 of the culture supernatant 15A in the flow path 67. The flow path 67 is an example of the "flow path" and "second flow path" according to the technology of the present invention. Furthermore, the flow direction FD2 is an example of the "fluid flow direction in the second flow path" according to the technology of the present invention.

[0048] With the flange 54 abutting against its bottom surface 55, the mounting hole 37 is formed at a depth where the center of the flow path 30 coincides with the center of the flow path 67. Then, with the rotation limiting member 46 not installed in the mounting hole 37, the probe 45 is rotated by the operator about the optical axis OA1, aligning its orientation with the flow direction FD2 coinciding with the flow direction FD1. Thus, the centers of the flow path 30 and flow path 67 are aligned, and the flow direction FD1 coincides with the flow direction FD2. Therefore, the line L1 connecting the center of the inlet 33 and the center of the outlet 34 of the flow path 30 and the line L2 connecting the center of the inlet 65 and the center of the outlet 66 of the flow path 67 are aligned (see reference). Figure 5 The term "consistency" as used here also refers not only to complete consistency, but also to consistency in the sense of an error that is generally permissible in the technical field to which the present invention pertains and does not deviate from the technical spirit of the present invention. The error referred to here is preferably ±10%, more preferably ±5%. More specifically, consistency between line L1 and line L2 means, for example, allowing an angular deviation of ±3°. Furthermore, the centers of flow path 30 and flow path 67 do not necessarily have to be aligned.

[0049] The rotation limiting member 46 has a large-diameter portion 70 at its base and a small-diameter portion 71 at its front end. A circular insertion hole 72 is formed at the center of the interior of the rotation limiting member 46. The diameter of the insertion hole 72 is larger than the diameter of the probe 45. Furthermore, the diameter of the insertion hole 72 decreases by one size at the boundary between the large-diameter portion 70 and the small-diameter portion 71. The probe 45 is inserted into the insertion hole 72. Therefore, the probe 45 forms the inner cylinder, and the rotation limiting member 46 forms the outer cylinder.

[0050] A thread 73 is cut into the small diameter portion 71. The thread 73 engages with the thread 39 of the mounting hole 37. Through these threads 39 and 73, the rotation limiting member 46 can be installed and removed relative to the flow pool 10.

[0051] When the rotation limiting member 46 is installed in the flow cell 10 via the mounting hole 37, the front end portion 74 of the small diameter portion 71 of the rotation limiting member 46 (see reference) Figure 4 and Figure 5 Pressed against flange portion 54 (reference) Figure 5 The flange 54 is held by the bottom surface 55 of the mounting hole 37 and the front end 74 of the small diameter portion 71 of the rotation limiting member 46. Thus, the rotation limiting member 46 restricts the rotation of the probe 45 about the optical axis OA1.

[0052] A through hole 75 is formed at the position of the large-diameter portion 70 corresponding to the groove 51 of the probe 45. The diameter of the through hole 75 is larger than the diameter of the bolt 53. The bolt 53 is inserted into the through hole 75.

[0053] The connecting member 47 has a large-diameter portion 80, a medium-diameter portion 81, and a small-diameter portion 82 sequentially from the base end side. The diameter of the large-diameter portion 80 is larger than the diameter of the through hole 72 in the large-diameter portion 70 of the rotation limiting member 46. The diameter of the medium-diameter portion 81 is slightly smaller than the diameter of the through hole 72 in the large-diameter portion 70 of the rotation limiting member 46. Furthermore, the diameter of the small-diameter portion 71 is slightly smaller than the diameter of the through hole 72, which decreases in size at the boundary between the large-diameter portion 70 and the small-diameter portion 71. Therefore, the connecting member 47 is inserted into the space between the probe 45 and the rotation limiting member 46 with a certain gap between it and the rotation limiting member 46. When the connecting member 47 is inserted into the space between the probe 45 and the rotation limiting member 46, in the radial direction, the medium-diameter portion 81 faces the large-diameter portion 70 of the rotation limiting member 46, and the small-diameter portion 82 faces the small-diameter portion 71 of the rotation limiting member 46.

[0054] A circular mounting hole 83 is formed at the center of the connecting component 47. A thread 84 is cut into the inner wall of the mounting hole 83. The mounting hole 83 and the fitting hole 85 (see reference) Figure 4 and Figure 5 The mating hole 85 is also a circular hole, and its center coincides with that of the mounting hole 83. The diameter of the mating hole 85 is smaller than that of the mounting hole 83.

[0055] The diameter of the fitting hole 85 is the same as the diameter of the probe 45, and the probe 45 fits into the fitting hole 85. More specifically, the diameter of the probe 45 and the fitting hole 85 is, for example, 12.1 mm. 12.1). Furthermore, the fitting tolerance of probe 45 is, for example, H7 (0 to +18 μm), and the fitting tolerance of fitting hole 85 is, for example, G6 (+6 μm to +17 μm). By fitting probe 45 into fitting hole 85, in other words, by fitting probe 45 into connecting member 47, the optical axis OA1 of optical system 60 and the optical axis OA2 of optical system 100 built into measuring head 25 (both referenced) are... Figure 6 Align the optical axis.

[0056] A threaded hole 86 is formed at the position of the middle diameter portion 81 corresponding to the groove 51 of the probe 45 and the through hole 75 of the rotation limiting member 46. The threaded hole 86 is a through hole, and when the probe 45 is fitted into the connecting member 47, the bottom surface 52 of the groove 51 of the probe 45 is exposed. The thread 87 cut at the front end of the bolt 53 is screwed into the threaded hole 86. A hexagonal hole for inserting a hex wrench is formed on the head of the bolt 53. After the probe 45 is fitted into the connecting member 47, the operator inserts the bolt 53 through into the through hole 75, and the thread 87 is screwed into the threaded hole 86. At this time, the probe 45 and the connecting member 47 are fixed by the front end of the bolt 53 pressing against the bottom surface 52 of the groove 51 of the probe 45. In addition, the bolt 53 can be made of metal or resin.

[0057] The fitting hole 85 has a length approximately the same as the length of the main body portion 48 of the probe 45, which is located higher than the flange portion 54. Then, when the connecting member 47 is inserted into the space between the probe 45 and the rotation limiting member 46, and the probe 45 is fitted into the connecting member 47, the front end of the middle diameter portion 81 abuts against the bottom surface of the through hole 72 in the large diameter portion 70 of the rotation limiting member 46. Furthermore, the front end of the small diameter portion 71 abuts against the upper surface of the flange portion 54.

[0058] The measuring head 25 has a linear and circular optical path 90 at its internal center. Similar to the optical path 50 of the probe 45, the excitation light EL and the Raman scattered light RSL pass through the optical path 90. The center of the optical path 90 is aligned with the optical axis OA2 of the optical system 100 built into the measuring head 25.

[0059] The measuring head 25 has a main body 91 and a front end 92. The diameter of the front end 92 is smaller than the diameter of the main body 91. The diameter of the front end 92 matches the diameter of the mounting hole 83 of the connecting member 47. Threads 93 are cut into the front end 92. The threads 93 engage with the threads 84 of the mounting hole 83 of the connecting member 47. The connecting member 47 and the unit 38 can be mounted and dismounted relative to the measuring head 25 through these threads 84 and 93.

[0060] As an example, such as Figure 6 As shown, an optical system 100 is built into the measuring head 25. The optical system 100 includes a collimating lens 101, a reflecting mirror 102, a dichroic filter 103, and a condenser lens 104.

[0061] The collimating lens 101 is positioned opposite the excitation optical fiber 105 laid on the cable 27. Excitation light EL, guided from the analyzer 26 by the excitation optical fiber 105, is incident on the collimating lens 101. The collimating lens 101 parallelizes the excitation light EL and directs it to the reflecting mirror 102. The reflecting mirror 102 reflects the parallelized excitation light EL toward the dichroic filter 103.

[0062] The dichroic filter 103 reflects the excitation light EL from the mirror 102 toward the optical system 60 of the probe 45. The excitation light EL is transmitted through the optical system 60 and focused to the focusing position FP. The focal length is determined by the diameter and refractive index of the hemispherical lens 61, and the focusing position FP is determined by the focal length. At this time, the focusing position FP is a point. The focusing position FP is located between the exit surface 63 of the transparent plate 62 and the inner wall surface of the bottom plate 64 of the front end 49. Ideally, the focusing position FP coincides with the point on the exit surface 63 of the transparent plate 62 that intersects with the optical axis OA1. That is, the focusing position FP is located on the exit surface 63 of the transparent plate 62.

[0063] The dichroic filter 103 transmits the Raman scattered light RSL captured by the optical system 60 and directs it to the condenser lens 104. The condenser lens 104 is positioned opposite the Raman scattered light optical fiber 106 laid on the cable 27. The condenser lens 104 focuses the Raman scattered light RSL from the dichroic filter 103 onto the Raman scattered light optical fiber 106. Furthermore, the optical axis OA2 of the optical system 100 is a line passing through the center of the dichroic filter 103 and the condenser lens 104.

[0064] Figure 7 The diagram shows a cross-sectional view of the lower part of the flow cell 10, the portion of the fitting hole 40 protruding from the front end portion 49, the downstream portion of the front end portion 49 (hereinafter referred to as other portions), and the narrowest portion 41 of the second transport path 19, cut in a plane orthogonal to the flow direction FD1. In the portion of the fitting hole 40, the flow path through which the culture supernatant 15A flows exists on both the inner and outer sides of the front end portion 49. In other words, the culture supernatant 15A flows through both the inner and outer sides of the front end portion 49. The flow path on the inner side of the front end portion 49 is the second flow path 67. The flow path on the outer side of the front end portion 49 is defined by the inner wall surface of the flow path 30 of the flow cell 10 and the outer wall surface of the front end portion 49. The area observed from the flow direction FD1 of the flow path of the culture supernatant 15A in the portion of the fitting hole 40 is denoted as A (in...). Figure 7 The area marked as "flow path area A of the protruding part at the front end" is shown in the middle.

[0065] In other locations, the culture supernatant 15A flows through the first flow path 30 itself. The area observed from the flow direction FD1 of this first flow path 30 is denoted as B (in...). Figure 7 (This is marked as "Flow path area B of other parts"). Furthermore, in the narrowest part 41, the flow path through which the culture supernatant 15A flows is the second transport path 19 itself. The area of ​​the second transport path 19, as observed from the flow direction FD1 of this narrowest part 41, is defined as C (in...). Figure 7 The area of ​​the narrowest part (C) is marked as "the flow path area at the narrowest point".

[0066] Similar to the portion of the fitting hole 40 when viewed from the upstream side of the flow path 30 at the front end 49, the narrowest portion 41 constitutes a flow-constriction section when viewed from the downstream side of the flow path 30 at the fitting hole 40. The difference BC between the area B of the flow path 30 of the flow cell 10 and the area C of the narrowest portion 41 is preset so that the damage to the antibody 17 and the like caused by the eddies generated at the narrowest portion 41 is within an acceptable range. Therefore, in the portion of the fitting hole 40, similar to the narrowest portion 41, the condition under which the damage to the antibody 17 and the like caused by the eddies is within an acceptable range can be expressed by the following equation (1).

[0067] BA≤BC (1) If we rearrange equation (1), then the condition under which the damage of the eddy current to antibody 17, etc., in the location of the interlocking hole 40 is within the allowable range can ultimately be represented by the following equation (2).

[0068] A≥C (2) That is, in order to keep the damage of the eddy current to the antibody 17 and the like within an acceptable range in the part of the interlocking hole 40, the front end 49 can have the following structure, that is, when viewed from the flow direction FD1, the area A of the flow path in the part of the interlocking hole 40 is greater than or equal to the area C of the narrowest part 41.

[0069] Next, the function of the above structure will be explained. First, the operator inserts the probe 45 into the mounting hole 37 of the flow cell 10 and fits the probe 45 into the fitting hole 40, thus positioning the probe 45 within the flow cell 10. The lower surface of the flange portion 54 of the probe 45 abuts against the bottom surface 55 of the mounting hole 37. As a result, the centers of the flow path 30 and the flow path 67 are aligned.

[0070] Next, the operator rotates probe 45 around optical axis OA1, aligning the direction of probe 45 so that the flow direction FD2 of culture supernatant 15A in flow path 67 of probe 45 is consistent with the flow direction FD1 of culture supernatant 15A in flow path 30 of flow cell 10. By performing these steps, the line L1 connecting the center of inlet 33 and outlet 34 of flow path 30 and the line L2 connecting the center of inlet 65 and outlet 66 of flow path 67 are aligned.

[0071] Next, the operator screws the thread 73 of the small diameter portion 71 of the rotation limiting member 46 into the thread 39 of the mounting hole 37 of the flow pool 10, and installs the rotation limiting member 46 into the mounting hole 37. At this time, the front end portion 74 of the small diameter portion 71 of the rotation limiting member 46 presses against the flange portion 54 of the probe 45, thus limiting the rotation of the probe 45 around the optical axis OA1.

[0072] When the rotation limiting member 46 is installed in the flow cell 10, a space is formed between the probe 45 and the rotation limiting member 46. The operator inserts the connecting member 47 into this space. As a result, the main body portion 48 of the probe 45, which is located higher than the flange portion 54, is fitted into the fitting hole 85 of the connecting member 47. In this way, by fitting the probe 45 into the connecting member 47, the optical axis OA1 of the optical system 60 is aligned with the optical axis OA2 of the optical system 100.

[0073] The operator inserts bolt 53 into the through hole 75 of the rotation limiting component 46. Then, the thread 87 of bolt 53 is screwed into the threaded hole 86 of the connecting component 47. At this time, the probe 45 and the connecting component 47 are fixed by the front end of bolt 53 pressing against the bottom surface 52 of the groove 51 of probe 45.

[0074] Finally, the operator installs the measuring head 25 onto the connecting member 47 by screwing the thread 93 of the front end 92 of the measuring head 25 into the thread 84 of the mounting hole 83 of the connecting member 47. Furthermore, the first transport path 18 is connected to the first connecting part 31, and the second transport path 19 is connected to the second connecting part 32. Thus, the measurement system 2, consisting of the flow cell 10 and the Raman spectrometer 11, is assembled into the cell culture section 12. Additionally, the measuring head 25 can be installed onto the connecting member 47 before inserting the connecting member 47 into the space between the probe 45 and the rotation limiting member 46. Furthermore, the first transport path 18 and the first connecting part 31, and the second transport path 19 and the second connecting part 32 can be connected before placing the probe 45 into the flow cell 10.

[0075] The culture supernatant 15A obtained from the culture tank 13 in the antibody production cells 16 flows in the flow path 30 of the flow cell 10. The culture supernatant 15A flows into the flow path 67 from the inlet 65 and flows out of the flow path 67 from the outlet 66.

[0076] The front end portion 49 has the following structure: when viewed from the flow direction FD1, the area A of the flow path through which the culture supernatant 15A flows in the portion of the interlocking hole 40 is greater than or equal to the area C of the narrowest portion 41 through which the culture supernatant 15A flows in the downstream side of the front end portion 49, where the area is narrowest when viewed from the flow direction FD1. Therefore, damage to antibodies 17 and the like from the eddies in the portion of the interlocking hole 40 can be controlled within an acceptable range.

[0077] Within the front end 49, the culture supernatant 15A is irradiated with excitation light EL, which passes through the excitation optical fiber 105, optical system 100, optical path 90, optical path 50, and optical system 60. The excitation light EL is focused at the focusing position FP by the optical system 60.

[0078] Raman scattered light (RSL) is generated through the interaction between excitation light (EL) and antibody 17 in the culture supernatant 15A. The RSL is captured by optical system 60 and output to analyzer 26 via optical paths 50 and 90, optical system 100, and Raman scattered light fiber 106. The RSL is converted into Raman spectral data 28 by analyzer 26.

[0079] like Figure 3 As shown, the narrowest portion 41 exists in the second transport path 19. Conversely, the narrowest portion 41 does not exist in the flow path 30 of the flow pool 10. Therefore, the possibility of eddies being generated in the flow path 30 of the flow pool 10, except for the portion with the fitting hole 40, can be reduced.

[0080] like Figure 3 As shown, a flow path 67 for the culture supernatant 15A is formed at the front end 49. Therefore, Raman spectral data 28 of the culture supernatant 15A can be measured within the front end 49. Furthermore, the front end 49 does not obstruct the flow of the culture supernatant 15A in the flow cell 10.

[0081] like Figure 5 As shown, the flow direction FD2 of the culture supernatant 15A in the flow path 67 of probe 45 is consistent with the flow direction FD1 of the culture supernatant 15A in the flow path 30 of flow cell 10. Therefore, the culture supernatant 15A flows smoothly in the flow path 30 and the flow path 67 without pressure loss, thereby reducing damage to antibodies such as antibody 17 in the culture supernatant 15A.

[0082] like Figure 3 As shown, the flow path 67 has an inlet 65 and an outlet 66 for the culture supernatant 15A, which is surrounded by a wall. Therefore, the flow of the culture supernatant 15A near the focusing position FP of the excitation light EL can be stabilized. Reduced component deviation in the culture supernatant 15A near the focusing position FP improves the stability of the Raman spectral data 28 measurement.

[0083] Raman scattering (RSL) readily reflects information about the functional groups of amino acids derived from proteins. Therefore, as shown in this example, by setting the physical property data to Raman spectral data 28, it is possible to obtain physical property data that more accurately reflects the concentration of antibody 17 as a protein.

[0084] Biopharmaceuticals containing antibody 17 as a cell product are called antibody pharmaceuticals, and are widely used not only for the treatment of chronic diseases such as cancer, diabetes, and rheumatoid arthritis, but also for the treatment of rare diseases such as hemophilia and Crohn's disease. Therefore, based on this example, where the culture supernatant 15A obtained from the culture tank 13 in the cultured antibody production cells 16 serves as the source of the antibody pharmaceutical, the development of antibody pharmaceuticals that can be widely used for the treatment of various diseases can be facilitated.

[0085] The narrowest part 41 can also exist in the flow path 30 of the flow pool 10.

[0086] The connecting component 47 can be non-removably mounted to the measuring head 25. That is, the measuring head 25 and the connecting component 47 can be integrated. Furthermore, the measuring head 25 and the probe 45 can be integrated.

[0087] The method for restricting the rotation of probe 45 about optical axis OA1 is not limited to the example of pressing the front end 74 of rotation restricting member 46 against the flange 54 of probe 45. For example, it could also be a method of fitting a protrusion formed on the front end 74 of rotation restricting member 46 into two or more fitting holes formed on flange 54. Furthermore, the method of fixing probe 45 and connecting member 47 is not limited to pressing the front end of bolt 53 against the bottom surface 52 of groove 51 of probe 45. It could also be a method of using a stop having a front end that can be protruded / retracted by spring force.

[0088] The connection between the flow cell 10 and the rotation limiting member 46, and the connection between the measuring head 25 and the connecting member 47, are secured by threads, but are not limited to this. Connections can also be made via spring-like claws that hang on the lower edge of the hole.

[0089] The flow path 67 of the probe 45 is not limited to a flow path having an inlet 65 and an outlet 66 surrounded by a wall, as illustrated in Japanese Patent Application Publication No. 2021-048872. It could also be a probe with an L-shaped front end in cross-section, as shown in Figures 13 and 19. Furthermore, the probe 45 may not have a flow path 67. For example, as described in Japanese Patent Application Publication No. 2020-511635, a portion of the optical system 60 may protrude into the flow path 30. In short, any probe can be configured with its front end 49 protruding from the flow path 30.

[0090] The inlet 65 and outlet 66 are not limited to a rectangular shape; they can also be circular, elliptical, or other shapes. Furthermore, the front end 49 is not limited to the illustrated cylindrical container shape; it can also be a square container shape, a hexagonal container shape, or other shapes. Therefore, the base plate 64 is not limited to the illustrated circular shape; it can also be rectangular, hexagonal, or other shapes. Furthermore, the peripheral plate is not limited to the illustrated curved surface; it can also be a plane.

[0091] The base plate 64, and consequently the inner wall surface of the base plate 64, can be a downwardly convex curved surface that mimics the shape of the flow path 30 of the flow cell 10. If the inner wall surface is set as a downwardly convex curved surface, the inner wall surface functions as a reflecting surface that directs the Raman scattered light RSL towards the optical system 60. Therefore, the S / N ratio of the Raman spectral data 28 can be further improved. Alternatively, the downwardly convex curved surface can also be in the shape of a parabolic antenna.

[0092] The lens constituting the optical system 60 is not limited to the illustrated hemispherical lens 61. It can also be a sphere, such as a spherical lens, a plano-convex lens, a biconvex lens, or a cylindrical lens. Furthermore, it is not limited to the illustrated circular transparent plate 62 with parallel incident and exit surfaces 63. It can also be a transparent plate with an incident surface and a planar exit surface, such as a spherical lens, a plano-convex lens, or a biconvex lens.

[0093] Alternatively, the lens and the transparent plate can be formed as a single lens, instead of constructing the optical system 60 by combining the lens and the transparent plate. Furthermore, the optical system 60 can be constructed without the transparent plate, or it can consist solely of the lens.

[0094] Alternatively, the first connecting portion 31 and the second connecting portion 32 can be disposed on the lower surface of the flow cell 10, and the flow path 30 can be U-shaped. The shape of the flow cell 10 is not limited to a cuboid shape, but can also be cylindrical or square. The cross-sectional shape of the flow path 30 is not limited to a circular shape, but can be elliptical or rectangular. When the flow path 30 is rectangular, the shape of the flow path 30 viewed from the flow direction FD1 can be such that the front end 49 is almost seamlessly embedded, and the size of the inlet 65 and the outlet 66 can be maximized so that the area of ​​the flow path 30 viewed from the flow direction FD1 is approximately the same as the area of ​​the flow path 67 (B≈A). Furthermore, the flow cell 10 and the unit 38 can be formed using composite materials such as carbon fiber reinforced resin.

[0095] The substances to be measured are not limited to antibody 17. They can also include proteins, peptides, nucleic acids (DNA, RNA (ribonucleic acid)), lipids, viruses, viral subunits, and virus-like particles, in addition to antibody 17.

[0096] Cell products are not limited to antibody 17. They can also be cytokines (interferon, interleukin, etc.) or hormones (insulin, glucagon, follicle-stimulating hormone, erythropoietin, etc.), growth factors (IGF (Insulin-Like Growth Factor)-1, bFGF (Basic Fibroblast Growth Factor), etc.), coagulation factors (Factors 7, 8, 9, etc.), enzymes (lysosomal enzymes, DNA (Deoxyribonucleic acid) degrading enzymes, etc.), Fc (Fragment Crystallizable) fusion proteins, receptors, albumins, and protein vaccines. Furthermore, antibody 17 also includes bispecific antibodies, antibody-drug conjugates, low-molecular-weight antibodies, and glycan-modified antibodies.

[0097] Physical property data are not limited to Raman spectroscopy data28. They can also be infrared absorption spectroscopy data, near-infrared absorption spectroscopy data, nuclear magnetic resonance spectroscopy data, ultraviolet-visible (UV-Vis) spectroscopy data, or fluorescence spectroscopy data.

[0098] The fluid is not limited to culture supernatant 15A. It can also be cell culture medium 15 before cells are removed by the cell removal filter 14. It can also be cell culture medium (so-called culture medium) that does not contain cell products before being supplied to the culture tank 13. It can also be a purified solution obtained by purifying culture supernatant 15A using a chromatographic apparatus in the purification section. The fluid is not limited to liquids related to cell culture, such as river water collected for investigating water pollution. It can also be a raw material (e.g., lithium polystyrene and methanol aqueous solution, etc.) and / or a product used in the continuous generation of monomers or polymers (e.g., polystyrene) through flow synthesis. Furthermore, the fluid is not limited to liquids and can also be a gas.

[0099] Based on the above records, one can master the techniques described in the following notes.

[0100] [Note 1] A probe is mounted in a flow cell having a first flow path through which fluid contains a substance for which the physical properties of a spectrophotometer are to be determined, and the probe is configured such that its distal end protrudes from the first flow path. The front end portion has the following structure: when viewed from the direction of fluid flow, the area of ​​the flow path through the protruding portion of the front end portion is greater than or equal to the area of ​​the narrowest portion of the flow path through which the fluid flows in the downstream side of the front end portion is narrowest when viewed from the direction of fluid flow.

[0101] [Note 2] According to the probe described in Appendix 1, wherein, The flow tank is connected to an inflow path that allows the fluid to flow into the first flow path from the outside and an outflow path that allows the fluid from the first flow path to flow out from the outside. The narrowest part exists in the outflow path.

[0102] [Note 3] The probe according to appendix 1 or 2, wherein, A second flow path through which the fluid flows is formed at the front end.

[0103] [Note 4] According to the probe described in Appendix 3, wherein, The flow direction of the fluid in the second flow path is the same as the flow direction of the fluid in the first flow path.

[0104] [Note 5] According to the probe described in Appendix 3 or 4, wherein, The second flow path has an inlet and an outlet for the fluid that are surrounded by walls.

[0105] [Note 6] The probe according to any one of notes 1 to 5, wherein, The physical property data are Raman spectral data.

[0106] [Note 7] The probe according to any one of notes 1 to 6, wherein, The fluid is any one of cell culture medium, culture supernatant, purified solution, and culture medium.

[0107] The technology of the present invention can also be appropriately combined with the various embodiments and / or variations described above. Furthermore, it is not limited to the embodiments described above; of course, various structures can be adopted as long as the spirit remains unchanged.

[0108] The descriptions and illustrations shown above are detailed explanations of the parts involved in the technology of this invention, and are merely examples of the technology of this invention. For example, the descriptions of the structure, function, effect, and effect described above are examples of the structure, function, effect, and effect of the parts involved in the technology of this invention. Therefore, it is natural that, without departing from the spirit of the technology of this invention, unnecessary parts may be deleted, new elements may be added, or substitutions may be made to the descriptions and illustrations shown above. Furthermore, in order to avoid complexity and to facilitate understanding of the parts involved in the technology of this invention, descriptions related to technical common sense that do not require special explanation have been omitted in the descriptions and illustrations shown above, based on the technology that enables the implementation of this invention.

[0109] In this specification, "A and / or B" has the same meaning as "at least one of A and B". That is, "A and / or B" means that it can be only A, only B, or a combination of A and B. Furthermore, in this specification, when three or more items are expressed in combination using "and / or", the same concept as "A and / or B" may also be applied.

[0110] All documents, patent applications and technical standards described in this specification are incorporated herein by reference to the same extent as when each document, patent application and technical standard was specifically and separately described.

Claims

1. A probe mounted to a flow cell having a first flow path through which a fluid containing a substance to be measured for physical property data by a spectroscopic analysis device is caused to flow, the probe being arranged in a state in which a front end portion thereof protrudes into the first flow path, wherein the front end portion has a structure in which, when viewed from the direction of flow of the fluid, the area of the flow path through which the fluid flows at the position where the front end portion protrudes is equal to or greater than the area of the flow path at the narrowest position where the area of the flow path viewed from the direction of flow of the fluid becomes narrowest on the downstream side of the front end portion.

2. The probe according to claim 1, wherein a flow-in path through which the fluid is caused to flow from the outside into the first flow path and a flow-out path through which the fluid from the first flow path is caused to flow to the outside are connected in the flow cell, and the narrowest position is present in the flow-out path.

3. The probe according to claim 1, wherein a second flow path through which the fluid flows is formed in the front end portion.

4. The probe according to claim 3, wherein the direction of flow of the fluid in the second flow path coincides with the direction of flow of the fluid in the first flow path.

5. The probe according to claim 3, wherein the second flow path has a flow inlet and a flow outlet of the fluid surrounded by a wall surface.

6. The probe according to claim 1, wherein the physical property data is Raman spectral data.

7. The probe according to claim 1, wherein the fluid is any one of a cell culture solution, a culture supernatant, a purified solution, and a culture medium. ​ ​ ​ ​ ​ ​ ​ ​

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