Probe

Abstract

This probe of a spectrum analyzer is attached to a flow cell having a first flow path through which flows a fluid containing a substance for which physical property data are to be measured. The probe is disposed in a state in which a tip part protrudes into the first flow path. The tip part has a configuration such that an area of a flow path through which the fluid flows at a location where the tip part protrudes, as viewed from a flow direction of the fluid, is equal to or larger than an area of a flow path at a narrowest location where an area viewed from the flow direction of the fluid becomes the narrowest in a flow path through which the fluid flows on a downstream side of the tip part.

Description

probe

[0001] The technology of the present disclosure relates to probes.

[0002] The spectroscopic analyzer has a probe. The tip of the probe has a built-in optical system for irradiating a measurement light onto a substance to be measured for physical property data and capturing return light from the substance to be measured. As described in JP-A-2020-511635, the probe (referred to as an optical analyzer connector in JP-A-2020-511635) is attached to a flow cell. JP-A-2020-511635 discloses an embodiment in which a ball lens as an optical system disposed at the tip of the probe protrudes into a flow path of a flow cell through which a fluid containing the substance to be measured flows.

[0003] As in the embodiment described in JP-A-2020-511635, when the tip of the probe protrudes into the flow path of the flow cell, the protruding portion becomes a flow constriction portion where the area of ​​the flow path as viewed from the direction of fluid flow is narrower than other portions. This flow constriction portion generates vortices, which may damage the substance to be measured in the fluid. However, JP-A-2020-511635 does not take measures to ensure that damage to the substance to be measured caused by the protruding portion is within an acceptable range.

[0004] One embodiment of the technique of the present disclosure provides a probe that can keep damage to a measurement target substance within an acceptable range.

[0005] The probe disclosed herein is a probe that is 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, and is positioned with its tip protruding into the first flow path, and the tip is configured so that the area of ​​the flow path through which the fluid flows at the point where the tip protrudes, as viewed from the direction of fluid flow, is equal to or greater than the area of ​​the flow path at the narrowest point downstream of the tip, which is the narrowest area as viewed from the direction of fluid flow.

[0006] The flow cell is connected to an inlet channel for allowing fluid to flow into the first flow channel from the outside and an outlet channel for allowing fluid to flow out from the first flow channel to the outside, and it is preferable that the narrowest point is located in the outlet channel.

[0007] The tip portion preferably has a second flow path formed therein through which a fluid flows.

[0008] It is preferable that the direction of fluid flow in the second flow path coincides with the direction of fluid flow in the first flow path.

[0009] The second flow path preferably has an inlet and an outlet for the fluid, the inlet and outlet being surrounded by a wall surface.

[0010] The physical property data is preferably Raman spectrum data.

[0011] The fluid is preferably any one of a cell culture medium, a culture supernatant, a purified liquid, and a medium.

[0012] According to the technology of the present disclosure, it is possible to provide a probe that can keep damage to a measurement target substance within an acceptable range.

[0013] FIG. 1 is a diagram showing how Raman spectral data of antibodies and the like in culture supernatant obtained in a cell culture section is measured by a measurement system. FIG. 2 is a diagram showing excitation light and Raman scattered light. FIG. 3 is an exploded perspective view of a flow cell, a spectroscopic analysis probe unit, and a measurement head. FIG. 4 is an exploded cross-sectional view of a flow cell, a spectroscopic analysis probe unit, and a measurement head. FIG. 5 is a cross-sectional view of a flow cell, a spectroscopic analysis probe unit, and a measurement head. FIG. 6 is a diagram showing an optical system built into the measurement head. FIG. 7 is an explanatory diagram showing the relationship in size between the flow channel areas of various parts of the flow cell and the narrowest part of the second delivery path.

[0014] As an example, as shown in Figure 1, the measurement system 2 includes a flow cell 10 and a Raman spectrometer 11. The measurement system 2 is incorporated into a cell culture unit 12 in a manufacturing system for active pharmaceutical ingredients for biopharmaceuticals, for example. The cell culture unit 12 has a culture tank 13 and a cell removal filter 14. A cell culture solution 15 is stored in the culture tank 13. The Raman spectrometer 11 is an example of a "spectroscopic analysis device" according to the technology of the present disclosure.

[0015] Antibody-producing cells 16 are seeded in the culture vessel 13, and the antibody-producing cells 16 are cultured in a cell culture solution 15. The antibody-producing cells 16 are cells established by incorporating an antibody gene into host cells such as Chinese hamster ovary cells (CHO cells). The antibody-producing cells 16 produce immunoglobulins, i.e., antibodies 17, during the culture process. Therefore, not only the antibody-producing cells 16 but also the antibodies 17 are present in the cell culture solution 15. The antibodies 17 are, for example, monoclonal antibodies, and are the active ingredients of biopharmaceuticals. The antibodies 17 are an example of a "substance to be measured" according to the technology of the present disclosure.

[0016] A first delivery path 18 is connected to the culture vessel 13. A cell removal filter 14 is disposed in the first delivery path 18. The cell removal filter 14 captures antibody-producing cells 16 in the cell culture solution 15 using a filter membrane (not shown), for example, by tangential flow filtration (TFF), thereby removing the antibody-producing cells 16 from the cell culture solution 15. The cell removal filter 14 also allows antibodies 17 to pass through. Therefore, the cell culture solution 15 containing mainly antibodies 17 flows downstream of the cell removal filter 14 in the first delivery path 18. The cell culture solution 15 from which the antibody-producing cells 16 have been removed by the cell removal filter 14 is referred to as a culture supernatant 15A. Hereinafter, the cell culture solution 15 from which the antibody-producing cells 16 have been removed by the cell removal filter 14 will be referred to as a culture supernatant 15A. The culture supernatant 15A is an example of a "fluid" according to the technology disclosed herein. In addition to the antibody 17, the culture supernatant 15A also contains cell-derived proteins, cell-derived DNA (deoxyribonucleic acid), aggregates of the antibody 17, viruses, etc. These cell-derived proteins, cell-derived DNA, aggregates of the antibody 17, and viruses are also examples of the "measurement target substance" according to the technology of the present disclosure. The cell removal filter 14 may be an alternating tangential flow filtration (ATF) filter.

[0017] The flow cell 10 is connected to a first delivery path 18 downstream of the cell removal filter 14. Culture supernatant 15A flows into the flow cell 10 from the first delivery path 18 at a preset flow rate. The first delivery path 18 is an example of a "flow path" and an "inflow path" according to the technology of the present disclosure. A delivery pump (not shown) is provided on the first delivery path 18 downstream of the cell removal filter 14 (between the cell removal filter 14 and the flow cell 10). The delivery pump delivers the culture supernatant 15A toward the flow cell 10 at a flow rate of 200 cc / min or more, for example, 300 cc / min.

[0018] A second outlet path 19 is also connected to the flow cell 10. The culture supernatant 15A that flows into the flow cell 10 from the first outlet path 18 flows out to the second outlet path 19. The second outlet path 19 is an example of a "flow path" and an "outlet path" according to the technology of the present disclosure. The second outlet path 19 is connected to a purification unit that purifies the antibody 17 from the culture supernatant 15A using, for example, a chromatography device, and sends the culture supernatant 15A from the flow cell 10 to the purification unit.

[0019] As an example, as shown in Figure 2, the Raman spectrometer 11 is an instrument that evaluates a substance M by utilizing the characteristics of Raman scattered light RSL. When excitation light EL is irradiated onto the substance M, the excitation light EL interacts with the substance M, generating Raman scattered light RSL having a different wavelength from that of the excitation light EL. The wavelength difference between the excitation light EL and the Raman scattered light RSL corresponds to the energy 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. Of the Stokes line and anti-Stokes line, it is preferable to use the Stokes line for the Raman scattered light RSL.

[0020] Returning to Figure 1, the Raman spectrometer 11 is composed of a measurement head 25 and an analyzer 26. The measurement head 25 is connected to the analyzer 26 via a cable 27. The measurement head 25 is attached to the flow cell 10. In the following description, the measurement head 25 side will be referred to as the "top" or "base end," and the flow cell 10 side will be referred to as the "bottom," "bottom," or "tip."

[0021] The analyzer 26 incorporates a light source that emits excitation light EL. The excitation light EL emitted from the light source is guided to the measurement head 25 via a cable 27. The measurement head 25 emits the excitation light EL from its tip. The excitation light EL is irradiated onto the culture supernatant 15A flowing within the flow cell 10. Raman scattered light RSL is generated by the interaction between this excitation light EL and the antibodies 17 and the like in the culture supernatant 15A. The measurement head 25 receives the Raman scattered light RSL. The Raman scattered light RSL received by the measurement head 25 is output to the analyzer 26 via the cable 27.

[0022] The analyzer 26 resolves the Raman scattered light RSL into wavenumbers and derives the intensity value of the Raman scattered light RSL for each wavenumber, thereby generating Raman spectrum data 28. The Raman spectrum data 28 is data in which the intensity value of the Raman scattered light RSL for each wavenumber is registered. In FIG. 1 , the Raman spectrum data 28 is －１ ~1800cm －１ The intensity values ​​of the Raman scattered light RSL in the range from 1 cm －１ The graph shown below the Raman spectrum data 28 is obtained by plotting the intensity values ​​of the Raman spectrum data 28 for each wavenumber and connecting them with a line. The Raman spectrum data 28 is an example of "physical property data" according to the technology of the present disclosure.

[0023] In this way, the measurement system 2 causes the culture supernatant 15A obtained from the culture tank 13 in which antibody-producing cells 16 are being cultured to flow through the flow cell 10. Then, the culture supernatant 15A flowing through the flow cell 10 is irradiated with excitation light EL via the measurement head 25, thereby measuring Raman spectrum data 28 of the antibodies 17 and the like in the culture supernatant 15A.

[0024] The Raman spectrum data 28 is used to predict the state of the substance to be measured, such as the concentration of antibody 17 in the culture supernatant 15A. In this case, for example, a machine learning model that outputs the concentration of antibody 17 in response to input of the Raman spectrum data 28 is utilized. Note that the concentration of aggregates in the culture supernatant 15A may also be predicted as the state of the substance to be measured. Furthermore, instead of or in addition to the concentration, density or the like may also be predicted.

[0025] As shown in Figures 3, 4, and 5 as an example, the flow cell 10 is a rectangular parallelepiped member having a linear flow path 30 with a circular cross section at its center. The flow path 30 is an example of the "flow path" and "first flow path" according to the technology of the present disclosure. The flow cell 10 is made of a metal such as Hastelloy. Alternatively, the flow cell 10 may be made of a resin such as a polyolefin resin. If made of a resin, the flow cell 10 may be single-use.

[0026] A cylindrical boss-shaped first connector 31 and a second connector 32 are provided on opposing end surfaces of the flow cell 10. The first connector 31 has an inlet 33 of the flow path 30, and the second connector 32 has an outlet 34 of the flow path 30. The direction parallel to the flow path 30 from the inlet 33 toward 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 "flow direction of the fluid in the first flow path" according to the technology of the present disclosure.

[0027] The first feed path 18 and the first connecting portion 31 are liquid-tightly connected by a ferrule joint. The ferrule joint is composed of ferrules (not shown) formed at one end of the first feed path 18 and one end of the first connecting portion 31, a gasket (not shown) sandwiched in a groove of the ferrule, and a first clamp 35 that secures one end of the first feed path 18 and one end of the first connecting portion 31. The second feed path 19 and the second connecting portion 32 are similarly liquid-tightly connected by a ferrule joint including a second clamp 36. Note that a parallel thread or a tapered thread may be used to connect the first feed path 18 and the first connecting portion 31 to the second feed path 19 and the second connecting portion 32.

[0028] The first delivery path 18 has the same diameter from the cell removal filter 14 to the end connected to the first connection part 31. The end of the first delivery path 18 connected to the first connection part 31 has a reverse tapered shape, gradually widening in diameter from the upstream side (the cell culture section 12 side) toward the downstream side (the flow cell 10 side) in the flow direction FD1. Conversely, the end of the second delivery path 19 connected to the second connection part 32 has a tapered shape, gradually narrowing in diameter from the upstream side (the flow cell 10 side) toward the downstream side (the purification section side) in the flow direction FD1. The second delivery path 19 maintains this narrowed diameter all the way to the purification section. The narrowed diameter point in the second delivery path 19 is the narrowest point 41, which is the smallest area, as viewed from the flow direction FD1, of the flow path through which the culture supernatant 15A flows, downstream of the tip 49 of the probe 45 (described later).

[0029] An attachment hole 37 is provided in the center of the top surface of the flow cell 10. The attachment hole 37 is a circular hole for detachably attaching a spectroscopic probe unit (hereinafter simply referred to as unit) 38 to the flow cell 10, and has a thread 39 on its inner wall surface. The attachment hole 37 is connected to a fitting hole 40 (see Figures 4 and 5). The fitting hole 40 is also a circular hole, and its center coincides with that of the attachment hole 37. The fitting hole 40 has a diameter slightly smaller than that of the attachment hole 37. The fitting hole 40 penetrates into the flow path 30.

[0030] The unit 38 is made up of a probe 45, a rotation restricting member 46, and a connecting member 47. The unit 38 is made of metal, for example, Hastelloy or the like. Alternatively, the unit 38 may be made of resin, for example, polyolefin resin. If the unit 38 is made of resin, it may be a single-use unit.

[0031] The probe 45 is a cylindrical member having a diameter that matches the diameter of the fitting hole 40. The operator of the measurement system 2 inserts the probe 45 into the mounting hole 37 and fits it into the fitting hole 40. This positions the probe 45 within the flow cell 10. When the rotation restricting member 46 is not attached to the mounting hole 37, but the probe 45 is simply inserted into the mounting hole 37 and fitted into the fitting hole 40, the probe 45 can rotate 360° in the circumferential direction. Here, "match" refers to not only perfect match, but also match that includes an error that is generally acceptable in the technical field to which the technology of the present disclosure belongs and that does not contradict the spirit of the technology of the present disclosure. The error here is preferably ±10%, and more preferably ±5%.

[0032] The probe 45 has a main body 48 and a tip 49. The main body 48 has a linear, circular optical path 50 at its center. The excitation light EL and the Raman scattered light RSL pass through the optical path 50. The excitation light EL passes through the optical path 50 from the measurement head 25 and the main body 48 toward the tip 49. Conversely, the Raman scattered light RSL passes through the optical path 50 from the tip 49 toward the main body 48 and ultimately the measurement head 25.

[0033] A radially recessed groove 51 is formed around the entire periphery of the outer peripheral surface of the upper portion of the main body 48. The tip of a bolt 53 is pressed against a bottom surface 52 of the groove 51 (see FIG. 5).

[0034] Furthermore, a flange 54 that protrudes radially is formed around the entire outer periphery of the central portion of the main body 48. When the probe 45 is inserted through the mounting hole 37 and fitted into the fitting hole 40, the lower surface of the flange 54 abuts against the bottom surface 55 (see FIGS. 4 and 5 ) of the mounting hole 37. In other words, the flange 54 functions as a stopper that prevents the main body 48 of the probe 45 above the flange 54 from slipping below the bottom surface 55.

[0035] A circular groove 56 (see FIGS. 4 and 5) is formed on the underside of the flange 54. An O-ring 57 is fitted into the groove 56. The O-ring 57 is made of elastic rubber. The O-ring 57 is compressed between the bottom surface 55 of the mounting hole 37 and the groove 56, thereby preventing the culture supernatant 15A flowing through the flow path 30 from leaking to the outside. Although not shown, O-rings for preventing leakage of the culture supernatant 15A are also disposed between the main body 48 and the tip 49, etc.

[0036] An optical system 60 is built into the lower part of the main body 48. The optical system 60 is disposed at a position where its optical axis OA1 (see FIGS. 4 and 5) coincides with the center of the optical path 50.

[0037] The optical system 60 is composed of a hemispherical lens 61 and a transparent plate 62. The hemispherical lens 61 is literally a hemispherical lens and is made of, for example, sapphire glass or quartz glass. The hemispherical lens 61 has a hemispherical entrance surface for the excitation light EL and a flat exit surface for the excitation light EL.

[0038] The transparent plate 62 is a circular plate having an incident surface and an exit surface 63 for the excitation light EL that are parallel to each other (see FIGS. 4 and 5 ), and is made of, for example, sapphire glass or quartz glass. Here, "parallel" refers to parallelism in the sense that it includes, in addition to perfect parallelism, an error that is generally acceptable in the technical field to which the technology of the present disclosure pertains and that does not contradict the spirit of the technology of the present disclosure. The error here is preferably ±10%, more preferably ±5%.

[0039] The curvature of the exit surface of the hemispherical lens 61 and the curvature of the entrance surface of the transparent plate 62 are the same (0 in this case). The exit surface of the hemispherical lens 61 and the entrance surface of the transparent plate 62 may be fixedly joined with an adhesive or the like, or may be held at the tip portion 49 in a state where the surfaces are simply joined together without using an adhesive or the like. Here, "same" refers to not only complete sameness, but also sameness in the sense of including an error that is generally acceptable in the technical field to which the technology of the present disclosure belongs and that does not contradict the spirit of the technology of the present disclosure. The error here is preferably ±10%, more preferably ±5%.

[0040] The tip portion 49 protrudes from the fitting hole 40 into the flow path 30. Therefore, when viewed from the flow path 30 upstream of the fitting hole 40, the location of the fitting hole 40 becomes a contracted flow section. The location of the fitting hole 40 is an example of a "location from which the tip portion protrudes" according to the technology of the present disclosure.

[0041] The tip portion 49 has a cylindrical container shape, with an open upper side connected to the main body portion 48, a flat bottom plate 64 (see FIGS. 4 and 5 ) at the lower side, and a peripheral plate extending upward from the bottom plate 64 and closed around the periphery. An inlet 65 and an outlet 66 for the culture supernatant 15A are formed at positions 180° symmetrical about the center of the peripheral plate. The inlet 65 and the outlet 66 are rectangular, or more precisely, square, and surrounded by a wall. The inlet 65 and the outlet 66, together with the space formed by the bottom plate 64 and the peripheral plate, form a flow path 67 (see FIGS. 4 and 5 ) through which the culture supernatant 15A flows. The direction parallel to the flow path 67 from the inlet 65 toward 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 a "flow path" and a "second flow path" according to the technology disclosed herein. Furthermore, the flow direction FD2 is an example of the "flow direction of the fluid in the second flow path" according to the technology of the present disclosure.

[0042] The mounting hole 37 is formed to a depth such that the center of the flow channel 30 coincides with the center of the flow channel 67 when the flange 54 abuts against its bottom surface 55. The probe 45, with the rotation restricting member 46 not attached to the mounting hole 37, is rotated around the optical axis OA1 by an operator to orient the probe 45 so that the flow direction FD2 coincides with the flow direction FD1. Since the centers of the flow channels 30 and 67 coincide and the flow directions FD1 and FD2 coincide, the line L1 connecting the center of the inlet 33 of the flow channel 30 and the center of the outlet 34 of the flow channel 67 coincides with the line L2 connecting the center of the inlet 65 of the flow channel 67 and the center of the outlet 66 of the flow channel 67 (see FIG. 5 ). The term "coincidence" as used herein refers not only to perfect coincidence but also to a degree of error generally acceptable in the technical field to which the technology of the present disclosure pertains, which does not contradict the spirit of the technology of the present disclosure. The error here is preferably ±10%, more preferably ±5%. More specifically, the line L1 and the line L2 being aligned means that an angular deviation of, for example, ±3° is permitted. Note that the centers of the flow channels 30 and 67 do not necessarily have to be aligned.

[0043] The rotation restricting member 46 has a large diameter portion 70 on the base end side and a small diameter portion 71 on the tip end side. A circular insertion hole 72 is formed in the center of the interior of the rotation restricting member 46. The insertion hole 72 is slightly larger in diameter than the probe 45. The diameter of the insertion hole 72 is smaller 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 serves as an inner cylinder, and the rotation restricting member 46 serves as an outer cylinder.

[0044] A screw 73 is formed in the small diameter portion 71. The screw 73 is threadedly engaged with the screw 39 of the mounting hole 37. The screw 39 and the screw 73 allow the rotation restricting member 46 to be attached to and detached from the flow cell 10.

[0045] When the rotation restricting member 46 is attached to the flow cell 10 via the mounting hole 37, the tip 74 (see FIGS. 4 and 5) of the small diameter portion 71 of the rotation restricting member 46 is pressed against the flange 54 (see FIG. 5). The flange 54 is sandwiched between the bottom surface 55 of the mounting hole 37 and the tip 74 of the small diameter portion 71 of the rotation restricting member 46. In this way, the rotation restricting member 46 restricts the rotation of the probe 45 around the optical axis OA1.

[0046] An insertion hole 75 is formed in the large diameter portion 70 at a position corresponding to the groove 51 of the probe 45. The diameter of the insertion hole 75 is larger than the diameter of the bolt 53. The bolt 53 is inserted into the insertion hole 75.

[0047] The connecting member 47 has, in order from the proximal end, a large diameter portion 80, a medium diameter portion 81, and a small diameter portion 82. The diameter of the large diameter portion 80 is larger than the diameter of the insertion hole 72 in the large diameter portion 70 of the rotation restricting member 46. The diameter of the medium diameter portion 81 is slightly smaller than the diameter of the insertion hole 72 in the large diameter portion 70 of the rotation restricting member 46. The diameter of the small diameter portion 71 is also slightly smaller than the diameter of the insertion hole 72, which is one step smaller 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 restricting member 46 with some backlash from the rotation restricting member 46. When the connecting member 47 is inserted into the space between the probe 45 and the rotation restricting member 46, the medium diameter portion 81 faces the large diameter portion 70 of the rotation restricting member 46, and the small diameter portion 82 faces the small diameter portion 71 of the rotation restricting member 46 in the radial direction.

[0048] A circular mounting hole 83 is formed in the center of the interior of the connecting member 47. A thread 84 is cut into the inner wall surface of the mounting hole 83. The mounting hole 83 is connected to a fitting hole 85 (see FIGS. 4 and 5). The fitting hole 85 is also a circular hole, and its center coincides with that of the mounting hole 83. The diameter of the fitting hole 85 is slightly smaller than that of the mounting hole 83.

[0049] The diameter of the fitting hole 85 matches the diameter of the probe 45, and the probe 45 is fitted into the fitting hole 85. More specifically, the diameter of the probe 45 and the fitting hole 85 is, for example, 12.1 mm (φ12.1). The fit tolerance of the probe 45 is, for example, H7 (0 to +18 μm), and the fit tolerance of the fitting hole 85 is, for example, G6 (+6 μm to +17 μm). When the probe 45 is fitted into the fitting hole 85, in other words, when the probe 45 is fitted into the connecting member 47, the optical axis OA1 of the optical system 60 and the optical axis OA2 of the optical system 100 built into the measuring head 25 (both see FIG. 6 ) are aligned.

[0050] A screw hole 86 is formed in the medium diameter portion 81 at a position corresponding to the groove 51 of the probe 45 and the insertion hole 75 of the rotation restricting member 46. The screw hole 86 is a through-hole that exposes the bottom surface 52 of the groove 51 of the probe 45 when the probe 45 is fitted to the connecting member 47. A screw 87 cut into the tip of a bolt 53 is threaded into the screw hole 86. A hexagonal hole is formed in the head of the bolt 53 for inserting a hexagonal wrench. After the probe 45 is fitted to the connecting member 47, the operator inserts the bolt 53 into the insertion hole 75 and threads the screw 87 into the screw hole 86. At this time, the tip of the bolt 53 is pressed against the bottom surface 52 of the groove 51 of the probe 45, thereby fixing the probe 45 and the connecting member 47. The bolt 53 may be made of metal or resin.

[0051] The fitting hole 85 has approximately the same length as the length of the main body 48 of the probe 45 above the flange 54. When the connecting member 47 is inserted into the space between the probe 45 and the rotation restricting member 46 and the probe 45 is fitted to the connecting member 47, the tip of the medium diameter portion 81 abuts against the bottom surface of the insertion hole 72 in the large diameter portion 70 of the rotation restricting member 46. The tip of the small diameter portion 71 abuts against the upper surface of the flange 54.

[0052] The measurement head 25 has a linear, circular optical path 90 at its center. Like 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 coincides with the optical axis OA2 of the optical system 100 built into the measurement head 25.

[0053] The measuring head 25 has a main body 91 and a tip 92. The tip 92 has a smaller diameter than the main body 91. The diameter of the tip 92 matches the diameter of the mounting hole 83 of the connecting member 47. A thread 93 is formed in the tip 92. The thread 93 screws into the thread 84 of the mounting hole 83 of the connecting member 47. The thread 84 and the thread 93 allow the connecting member 47, and therefore the unit 38, to be attached to and detached from the measuring head 25.

[0054] 6, the measurement head 25 includes an optical system 100. The optical system 100 includes a collimating lens 101, a mirror 102, a dichroic filter 103, and a condenser lens 104.

[0055] The collimating lens 101 is provided at a position facing the excitation light optical fiber 105 laid in the cable 27. The excitation light EL guided from the analyzer 26 by the excitation light optical fiber 105 is incident on the collimating lens 101. The collimating lens 101 collimates the excitation light EL and outputs it to the mirror 102. The mirror 102 reflects the collimated excitation light EL toward the dichroic filter 103.

[0056] 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 passes through the optical system 60 and is focused at a 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. In this case, the focusing position FP is point-like. 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 tip portion 49. Ideally, the focusing position FP coincides with the point where the exit surface 63 of the transparent plate 62 intersects with the optical axis OA1. In other words, the focusing position FP is located at the exit surface 63 of the transparent plate 62.

[0057] The dichroic filter 103 transmits the Raman scattered light RSL taken in by the optical system 60 and outputs it to the condenser lens 104. The condenser lens 104 is provided in a position facing the optical fiber 106 for Raman scattered light laid in the cable 27. The condenser lens 104 condenses the Raman scattered light RSL from the dichroic filter 103 into the optical fiber 106 for Raman scattered light. The optical axis OA2 of the optical system 100 is a line passing through the centers of the dichroic filter 103 and the condenser lens 104.

[0058] FIG. 7 shows a cross-sectional view of the lower portion of the flow cell 10, as well as a cross-sectional view taken along a plane perpendicular to the flow direction FD1 of the fitting hole 40 where the tip 49 protrudes, the downstream portion of the tip 49 (hereinafter referred to as "other portion"), and the narrowest portion 41 of the second delivery path 19. At the fitting hole 40, the flow path through which the culture supernatant fluid 15A flows exists both inside and outside the tip 49. In other words, the culture supernatant fluid 15A flows inside and outside the tip 49. The flow path inside the tip 49 is the second flow path 67. The flow path outside the tip 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 tip 49. The area of ​​the flow path for the culture supernatant fluid 15A at the fitting hole 40 as viewed in the flow direction FD1 is designated A (denoted as "flow path area A at the portion where the tip protrudes" in FIG. 7).

[0059] At other locations, the flow path through which the culture supernatant fluid 15A flows is the first flow path 30 itself. The area of ​​this first flow path 30 as viewed from the flow direction FD1 is designated B (indicated as "flow path area B at other locations" in FIG. 7). At the narrowest point 41, the flow path through which the culture supernatant fluid 15A flows is the second delivery path 19 itself. The area of ​​the second delivery path 19 as viewed from the flow direction FD1 at this narrowest point 41 is designated C (indicated as "flow path area C at narrowest location" in FIG. 7).

[0060] Similar to the location of the fitting hole 40 when viewed from the flow path 30 upstream of the tip 49, the narrowest point 41 is a contracted flow section when viewed from the flow path 30 downstream of the fitting hole 40. The difference B-C between the area B of the flow path 30 of the flow cell 10 and the area C of the narrowest point 41 is preset to a value that ensures that damage to the antibodies 17 and other components caused by vortex currents generated at the narrowest point 41 is within an acceptable range. Therefore, the condition for the damage to the antibodies 17 and other components caused by vortex currents at the fitting hole 40 to be within an acceptable range, just like at the narrowest point 41, is expressed by the following formula (1): B-A≦B-C (1)

[0061] Rearranging equation (1), the condition under which damage to antibodies 17 and the like due to vortex currents at fitting hole 40 is within an acceptable range is ultimately expressed by the following equation (2): A ≥ C (2) In other words, in order for damage to antibodies 17 and the like due to vortex currents at fitting hole 40 to be within an acceptable range, tip portion 49 should be configured so that the area A of the flow path at fitting hole 40 as viewed from flow direction FD1 is equal to or greater than the area C of narrowest point 41.

[0062] Next, the operation of the above configuration will be described. 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, thereby placing the probe 45 inside the flow cell 10. The lower surface of the flange 54 of the probe 45 abuts against the bottom surface 55 of the mounting hole 37. This aligns the centers of the flow channels 30 and 67.

[0063] Next, the operator rotates the probe 45 around the optical axis OA1 and orients the probe 45 so that the flow direction FD2 of the culture supernatant 15A in the flow path 67 of the probe 45 coincides with the flow direction FD1 of the culture supernatant 15A in the flow path 30 of the flow cell 10. By performing these procedures, the line L1 connecting the center of the inlet 33 and the center of the outlet 34 of the flow path 30 coincides with the line L2 connecting the center of the inlet 65 and the center of the outlet 66 of the flow path 67.

[0064] Next, the operator screws the thread 73 of the small diameter portion 71 of the rotation restricting member 46 into the thread 39 of the mounting hole 37 of the flow cell 10, and attaches the rotation restricting member 46 to the mounting hole 37. At this time, the tip 74 of the small diameter portion 71 of the rotation restricting member 46 is pressed against the flange 54 of the probe 45, thereby restricting rotation of the probe 45 around the optical axis OA1.

[0065] When the rotation restricting member 46 is attached to the flow cell 10, a space is formed between the probe 45 and the rotation restricting member 46. The operator inserts the connecting member 47 into this space. As a result, the main body 48 of the probe 45 above the flange 54 is fitted into the fitting hole 85 of the connecting member 47. By fitting the probe 45 into the connecting member 47 in this manner, the optical axes OA1 of the optical system 60 and OA2 of the optical system 100 are aligned.

[0066] The operator inserts the bolt 53 into the insertion hole 75 of the rotation restricting member 46. The operator also screws the thread 87 of the bolt 53 into the screw hole 86 of the connecting member 47. At this time, the tip of the bolt 53 is pressed against the bottom surface 52 of the groove 51 of the probe 45, thereby fixing the probe 45 and the connecting member 47 together.

[0067] Finally, the operator attaches the measuring head 25 to the connecting member 47 by threading the screw 93 on the tip 92 of the measuring head 25 into the screw 84 in the mounting hole 83 of the connecting member 47. The operator also connects the first outlet path 18 to the first connecting portion 31, and the second outlet path 19 to the second connecting portion 32. This allows the measurement system 2, consisting of the flow cell 10 and the Raman spectrometer 11, to be incorporated into the cell culture unit 12. The measuring head 25 may be attached to the connecting member 47 before inserting the connecting member 47 into the space between the probe 45 and the rotation restricting member 46. The operator may also connect the first outlet path 18 to the first connecting portion 31, and the second outlet path 19 to the second connecting portion 32 before placing the probe 45 in the flow cell 10.

[0068] A culture supernatant 15A obtained from a culture tank 13 in which antibody-producing cells 16 are being cultured flows through the flow path 30 of the flow cell 10. The culture supernatant 15A flows into the flow path 67 from an inlet 65 and flows out of the flow path 67 from an outlet 66.

[0069] The tip portion 49 is configured so that the area A of the flow path through which the culture supernatant 15A flows at the fitting hole 40 as viewed in the flow direction FD1 is equal to or larger than the area C of the flow path at the narrowest point 41, which is the narrowest area as viewed in the flow direction FD1 among the flow paths through which the culture supernatant 15A flows downstream of the tip portion 49. This makes it possible to keep damage to the antibody 17 and the like due to vortex flow at the fitting hole 40 within an acceptable range.

[0070] Within the tip portion 49, the culture supernatant 15A is irradiated with excitation light EL that has passed through the excitation light optical fiber 105, the optical system 100, the optical path 90, the optical path 50, and the optical system 60. The excitation light EL is focused by the optical system 60 at a focusing position FP.

[0071] Raman scattered light RSL is generated by the interaction between the excitation light EL and the antibodies 17 and the like in the culture supernatant 15A. The Raman scattered light RSL is taken in by the optical system 60, passes through the optical path 50, the optical path 90, the optical system 100, and the Raman scattered light optical fiber 106, and is output to the analyzer 26. The analyzer 26 converts the Raman scattered light RSL into Raman spectrum data 28.

[0072] 3 and other figures, the narrowest point 41 is present in the second delivery path 19. Conversely, the narrowest point 41 is not present in the flow path 30 of the flow cell 10. Therefore, it is possible to reduce the risk of vortex flow occurring in the flow path 30 of the flow cell 10 at a location other than the fitting hole 40.

[0073] 3 and other figures, a flow path 67 through which the culture supernatant 15A flows is formed in the tip 49. This allows Raman spectrum data 28 of the culture supernatant 15A to be measured within the tip 49. Furthermore, the tip 49 does not interfere with the flow of the culture supernatant 15A in the flow cell 10.

[0074] 5, the flow direction FD2 of the culture supernatant 15A in the flow path 67 of the probe 45 coincides with the flow direction FD1 of the culture supernatant 15A in the flow path 30 of the flow cell 10. This allows the culture supernatant 15A to flow smoothly through the flow paths 30 and 67 without causing pressure loss, reducing damage to the antibodies 17 and the like in the culture supernatant 15A.

[0075] 3 and other figures, the flow path 67 has an inlet 65 and an outlet 66 for the culture supernatant 15A, which are surrounded by a wall. This stabilizes the flow of the culture supernatant 15A near the focusing position FP of the excitation light EL. This reduces the concentration of components in the culture supernatant 15A near the focusing position FP, thereby improving the measurement stability of the Raman spectrum data 28.

[0076] Raman scattered light RSL is likely to reflect information derived from the functional groups of amino acids in proteins. Therefore, by using Raman spectrum data 28 as the physical property data as in this example, it is possible to obtain physical property data that better reflects the physical properties, such as the concentration of antibody 17, which is a protein.

[0077] Biopharmaceuticals containing antibody 17, which is a cell product, are called antibody drugs and are widely used to treat chronic diseases such as cancer, diabetes, and rheumatoid arthritis, as well as rare diseases such as hemophilia and Crohn's disease. Therefore, according to this example, in which culture supernatant 15A, which is the source of antibody drugs obtained from culture tank 13 in which antibody-producing cells 16 are cultured, is used as the fluid, it is possible to promote the development of antibody drugs that are widely used to treat various diseases.

[0078] The narrowest point 41 may be present in the flow path 30 of the flow cell 10 .

[0079] The connecting member 47 may be attached non-detachably to the measuring head 25. That is, the measuring head 25 and the connecting member 47 may be integrated. Also, the measuring head 25 and the probe 45 may be integrated.

[0080] The method for restricting the rotation of the probe 45 around the optical axis OA1 is not limited to the exemplified method of pressing the tip 74 of the rotation restricting member 46 against the flange 54 of the probe 45. For example, a method in which protrusions formed on the tip 74 of the rotation restricting member 46 are fitted into two or more fitting holes formed in the flange 54 may be used. Furthermore, the method for fixing the probe 45 and the connecting member 47 is not limited to the method in which the tip of the bolt 53 is pressed against the bottom surface 52 of the groove 51 of the probe 45. A method using a stopper having a tip that can be protruded and retracted by spring bias may also be used.

[0081] Although screw fastening is used to connect the flow cell 10 and the rotation restricting member 46, and the measuring head 25 and the connecting member 47, etc., the present invention is not limited to this. Connections may also be made using leaf spring-like claws that engage with the lower edges of holes.

[0082] The flow path 67 of the probe 45 is not limited to the example having an inlet 65 and an outlet 66 surrounded by a wall. It may also be a probe having a tip with an L-shaped cross section, as shown in FIGS. 13 and 19 of JP 2021-048872 A. The probe 45 may not have a flow path 67. For example, as in the embodiment described in JP 2020-511635 A, a portion of the optical system 60 may protrude into the flow path 30. In short, it is sufficient for the probe to be disposed with its tip 49 protruding into the flow path 30.

[0083] The inlet 65 and the outlet 66 are not limited to a rectangular shape but may be circular, elliptical, or the like. Furthermore, the tip 49 is not limited to the exemplified cylindrical container shape but may be a square or hexagonal cylindrical container shape. Therefore, the bottom plate 64 is also not limited to the exemplified circular shape but may be a rectangular or hexagonal shape. Furthermore, the peripheral plate is also not limited to the exemplified curved surface but may be a flat surface.

[0084] The bottom plate 64, and in turn the inner wall surface of the bottom plate 64, may be a downwardly convex curved surface that follows the shape of the flow channel 30 of the flow cell 10. If the inner wall surface is a downwardly convex curved surface, the inner wall surface serves as a reflective surface that directs the Raman scattered light RSL toward the optical system 60. This makes it possible to further increase the S / N ratio of the Raman spectrum data 28. The downwardly convex curved surface may also be in the shape of a parabolic antenna.

[0085] The lenses constituting the optical system 60 are not limited to the exemplified hemispherical lens 61. They may be spherical ball lenses, plano-convex lenses, biconvex lenses, cylindrical lenses, or the like. Furthermore, they are not limited to the exemplified circular transparent plate 62 having parallel entrance and exit surfaces 63. They may be transparent plates having an entrance surface shaped to match the shape of the exit surface of a ball lens, plano-convex lens, biconvex lens, or the like, and a flat exit surface.

[0086] The optical system 60 may not be formed by bonding a lens and a transparent plate, but may be formed integrally with the lens as a single lens. Also, the transparent plate may not be necessary, and the optical system 60 may be formed only with a lens.

[0087] The first connector 31 and the second connector 32 may be disposed on the underside of the flow cell 10, forming the flow path 30 in a U-shape. The shape of the flow cell 10 is not limited to a rectangular parallelepiped, but may also be cylindrical or rectangular. The cross-sectional shape of the flow path 30 is also not limited to a circular shape, but may be an elliptical or rectangular shape. When the flow path 30 is rectangular, the shape of the flow path 30 as viewed from the flow direction FD1 may be such that the tip 49 fits in with almost no gap, and the sizes of the inlet 65 and the outlet 66 may be as large as possible so that the area of ​​the flow path 30 as 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, the unit 38, and the like may be formed from a composite material such as carbon fiber reinforced resin.

[0088] The substance to be measured is not limited to the antibody 17, etc. Other substances than the antibody 17, such as proteins, peptides, nucleic acids (DNA, RNA (Ribonucleic Acid)), lipids, viruses, virus subunits, and virus-like particles, may also be used.

[0089] The cell product is not limited to antibody 17, etc. It may also be a cytokine (interferon, interleukin, etc.), a hormone (insulin, glucagon, follicle-stimulating hormone, erythropoietin, etc.), a growth factor (IGF (insulin-like growth factor)-1, bFGF (basic fibroblast growth factor), etc.), a blood coagulation factor (factor 7, factor 8, factor 9, etc.), an enzyme (lysosomal enzyme, DNA (deoxyribonucleic acid) degrading enzyme, etc.), an Fc (fragment crystalline) fusion protein, a receptor, albumin, or a protein vaccine. The antibody 17 also includes bispecific antibodies, antibody-drug conjugates, low molecular weight antibodies, sugar chain modified antibodies, and the like.

[0090] The physical property data is not limited to the Raman spectrum data 28. It may be infrared absorption spectrum data, near-infrared absorption spectrum data, nuclear magnetic resonance spectrum data, ultraviolet visible absorption spectroscopy (UV-Vis) spectrum data, or fluorescence spectrum data.

[0091] The fluid is not limited to the culture supernatant 15A. It may be the cell culture fluid 15 before the cells are removed by the cell removal filter 14. It may be a cell culture fluid (so-called culture medium) that does not contain cell products before being supplied to the culture tank 13. It may be a purified liquid obtained by purifying the culture supernatant 15A using a chromatography device in the purification unit. The fluid is not limited to a liquid related to cell culture, and may be, for example, river water collected to check for water pollution. It may also be a raw material (e.g., polystyryllithium and an aqueous methanol solution) and / or a product when continuously producing a product such as a monomer or polymer (e.g., polystyrene) by flow synthesis. Furthermore, the fluid is not limited to a liquid and may be a gas.

[0092] From the above description, the technology described in the following supplementary paragraphs can be understood.

[0093] [Supplementary Item 1] 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 analyzer flows, the probe being disposed with a tip protruding into the first flow path, wherein the tip is configured so that the area of ​​the flow path through which the fluid flows at the point where the tip protrudes, as viewed in the direction of fluid flow, is equal to or greater than the area of ​​a narrowest point of the flow path through which the fluid flows downstream of the tip, the narrowest point being the area of ​​the flow path through which the fluid flows downstream of the tip. [Supplementary Item 2] The flow cell is connected to an inflow path through which the fluid flows from the outside into the first flow path and an outflow path through which the fluid flows from the first flow path to the outside, the narrowest point being present in the outflow path. [Supplementary Item 3] The probe according to Supplementary Item 1 or Supplementary Item 2, wherein the tip is formed with a second flow path through which the fluid flows. [Supplementary Item 4] The probe according to Supplementary Item 3, wherein the tip is formed with a second flow path through which the fluid flows. [Supplementary Item 4] The probe according to Supplementary Item 3, wherein the flow direction of the fluid in the second flow path and the flow direction of the fluid in the first flow path are the same. [Supplementary Item 5] The probe according to Supplementary Item 3 or Supplementary Item 4, wherein the second flow path has an inlet and an outlet for the fluid surrounded by a wall. [Supplementary Item 6] The probe according to any one of Supplementary Items 1 to 5, wherein the physical property data is Raman spectrum data. [Supplementary Item 7] The probe according to any one of Supplementary Items 1 to 6, wherein the fluid is any one of a cell culture medium, a culture supernatant, a purified solution, and a culture medium.

[0094] The technology of the present disclosure can be appropriately combined with the various embodiments and / or various modified examples described above. Furthermore, it is needless to say that the technology is not limited to the above-described embodiments, and various configurations can be adopted as long as they do not deviate from the gist of the present disclosure.

[0095] The above-described description and illustrations are a detailed explanation of the parts related to the technology of the present disclosure and are merely an example of the technology of the present disclosure. For example, the above description of the configuration, functions, actions, and effects is an explanation of an example of the configuration, functions, actions, and effects of the parts related to the technology of the present disclosure. Therefore, it goes without saying that unnecessary parts may be deleted, new elements may be added, or replacements may be made to the above-described description and illustrations within the scope of the gist of the technology of the present disclosure. Furthermore, to avoid confusion and facilitate understanding of the parts related to the technology of the present disclosure, the above-described description and illustrations omit explanations of common technical knowledge that do not require particular explanation to enable the implementation of the technology of the present disclosure.

[0096] In this specification, "A and / or B" is synonymous with "at least one of A and B." In other words, "A and / or B" means that it may be only A, only B, or a combination of A and B. Furthermore, in this specification, the same concept as "A and / or B" is also applied when three or more things are expressed by connecting them with "and / or."

[0097] All publications, patent applications, and technical standards mentioned in this specification are herein incorporated by reference to the same extent as if each individual publication, patent application, or technical standard was specifically and individually indicated to be incorporated by reference.

Claims

1. A probe that is 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 instrument flows, the probe being positioned with its tip protruding into the first flow path, wherein the tip is configured so that the area of ​​the flow path through which the fluid flows at the point where the tip protrudes, as viewed in the direction of flow of the fluid, is greater than or equal to the area of ​​the flow path at the narrowest point downstream of the tip, which is the narrowest in area as viewed in the direction of flow of the fluid.

2. A probe as described in claim 1, wherein the flow cell is connected to an inflow path for allowing the fluid to flow into the first flow path from the outside, and an outflow path for allowing the fluid to flow out from the first flow path to the outside, and the narrowest point is present in the outflow path.

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

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

5. The probe according to claim 3, wherein said second flow path has an inlet and an outlet for said fluid, the inlet and outlet being surrounded by a wall.

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

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

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