Reaction vessel with stator insert and rotor

Abstract

The reaction vessel comprises a housing having a first end for connection to the lid and an opposing second end closed by a bottom wall, a rib inside the housing for a central opening adapted to receive the stator, the stator arranged inside the housing, a rotor having a cylindrical portion arranged within the stator, and a shaft having a first end to which a magnetic shaft drive is fixed and an opposing second end covered by and fixed to the rotor, the shaft extending into a first bearing of the bearing block, the first bearing being arranged on the shaft at an axial portion arranged between the magnetic shaft drive and the rotor, preferably the first end of the shaft is a free end and arranged within a recess in the lid, the free end of the shaft and the recess in the lid forming a second bearing which is a friction bearing, the shaft, the magnetic shaft drive, the first bearing and the second bearing from which the shaft extends, the rotor, the stator, preferably an extension of the stator, and / or the detection unit being arranged coaxially around the longitudinal axis of the housing.

Description

[Technical Field]

[0001] The present invention relates to a device, which is a reaction vessel suitable for applying a predetermined shear force to a liquid sample, a method for manufacturing the reaction vessel, and a process for using the device for applying a predetermined shear force to a sample, preferably including optical analysis of the sample passing through a portion of the reaction vessel. [Background technology]

[0002] The reaction vessel comprises a rotor extending on a bearing, optionally on just one bearing, the rotor having a stem with a drive part at one end and adapted to receive rotational torque from a drive motor which can be located adjacent to the drive part or at a distance from the drive part. Preferably, the bearing is located at a distance from the level of the liquid sample placed in the vessel so that the bearing does not come into contact with the liquid.

[0003] The reactor vessel includes a stator coaxially disposed with a rotor, and a predetermined gap is formed between the rotor and the stator, which allows a predetermined shear stress to be applied to the liquid, the shear stress being proportional to the rotational frequency of the rotor, the radius of the rotor, and the viscosity of the liquid, and the shear stress being inversely proportional to the gap width between the rotor and the stator.

[0004] The reaction vessel allows for circulation of the liquid sample driven by the rotation of the rotor, which acts as a centrifugal pump element. The vessel geometry is designed so that the maximum shear stress is applied to the liquid at the section with the smallest gap width between the rotor and stator.

[0005] The reaction vessel preferably comprises a detection portion located at the lower end of the reaction vessel, the detection portion being at least partially optically transmissive, e.g., for light to illuminate the interior volume of the reaction vessel and for detecting radiation passing through and / or emitted from the interior volume of the reaction vessel.

[0006] The reaction vessel has the advantage that it can be manufactured by arranging the elements of the reaction vessel in a manner predetermined by the shape of the elements. All of the elements of the reaction vessel, except for some of the elements of the reaction vessel, in particular the magnet and preferably the drive shaft, can be manufactured by injection molding of a synthetic resin. Thus, the reaction vessel can be manufactured from a single component, preferably manufactured by injection molding of a synthetic resin, by arranging the elements for a friction fit and / or a form fit, preferably by moving the elements of the reaction vessel relative to each other for a friction fit until they abut against a stop surface.

[0007] prior art Patent Document 1 (WO2012 / 110570A1) shows a schematic diagram of a reaction vessel in which a rotor extends on a mandrel that extends over a bearing located on a lid covering the vessel and a second bearing located at the bottom of the lid, the rotor having a conical shape parallel to the conically tapering bottom of the vessel. In an alternative example, the vessel extends on a sleeve located in the upper half of a cylindrical vessel, and the rotor extends into a tube that is open at both ends, the tube being located in the lower half of the vessel at a distance from the rotor and from the vessel bottom.

[0008] Patent Document 2 (WO2016 / 001334A1) shows a reaction vessel in which a stator is inserted at a distance from the vessel wall and at a distance from a central rotor. The rotor is cylindrical with a chamfered periphery. The stator has an extension below the portion surrounding the rotor, and the light source and detector are oriented at 90° to each other and 45° to a common central axis and are directed into the space formed by the extension. [Prior art documents] [Patent documents]

[0009] [Patent Document 1] International Publication No. 2012 / 110570 [Patent Document 2] International Publication No. 2016 / 001334 Summary of the Invention [Problem to be solved by the invention]

[0010] It is an object of the present invention to provide an alternative reaction vessel suitable for applying shear stress to a liquid, and an alternative process for applying shear force to a liquid by using a reaction vessel. Preferably, the object is to provide a reaction vessel assembled to effectively apply shear force to the entire volume of liquid contained therein, and to provide a vessel that allows optical detection of a representative portion of the liquid, the vessel being assembled from elements that can be manufactured separately, for example by plastic injection molding, a magnet as part of the drive, and a metal drive shaft. [Means for solving the problem]

[0011] The present invention achieves this object by the features of the claims, specifically by providing a reaction vessel suitable for use in applying a shear force to the entire volume of a liquid contained therein, the vessel being constructed for circulating a liquid therethrough, the vessel comprising: - a housing extending along a central longitudinal axis, the housing having a first end for connection to a lid and an opposing second end closed by a bottom wall, and at least one window portion, e.g. a detection portion of the housing wall having preferably two window portions arranged opposite each other or arranged at an angle of 90° to each other, arranged at the second end, the window portion being optically transparent and preferably flat, the detection portion preferably arranged adjacent to the bottom wall; the housing includes at least three ribs extending along the housing wall to the interior of the housing and projecting toward the longitudinal axis; the rib adjacent the first end of the housing has a first portion, the rib adjacent the first portion has a second portion, the second portion of the rib extending toward the longitudinal axis to define a central opening about the longitudinal axis, the central opening adapted to receive the stator; a first portion of at least one of the ribs, and preferably the first portion of each rib, extends further toward the longitudinal axis than a second portion thereof; The ribs have a stop surface formed between a first portion and a second portion of at least one rib, and preferably the first portion and the second portion of each rib are respectively disposed along a common straight line parallel to the longitudinal axis. Housing and - a stator disposed within the housing, the stator having a cylindrical inner surface for receiving the rotor at a fixed interval, the cylindrical surface having a first open end cross-section and a second end opening, e.g., a second open end cross-section, opposite the first open end cross-section, the stator outer surface being spaced apart from the housing; the form-fitted and / or friction-fitted stator is positioned such that a first end of the stator abuts a stop surface formed between the first portion and the second portion of the at least one rib; the stator is disposed in a friction fit and / or form fit between the second portions of the at least three ribs; the ribs, the stator and the housing wall define an outer channel of the stator, the channel having an open end cross section at each end thereof, preferably the outer peripheral surface of the stator is cylindrical, more preferably the end cross section of the stator comprises a ring-shaped surface perpendicular to the central longitudinal axis, the surface optionally being chamfered; - optionally, the stator having the second end cross-sectional opening is positioned directly adjacent to or spaced apart from the sensing portion; - optionally, the stator at its second end has an opening extending across the entire cross section or has a terminal portion tapering to an opening opening to the detection portion, the opening terminating directly adjacent to the detection portion or spaced apart from the detection portion; a stator; - a rotor having a cylindrical portion disposed within a stator or consisting of a cylindrical portion disposed within a stator, the rotor and stator being spaced apart by a ring-shaped gap of constant radius, the rotor having a second end covering the second end of the shaft, the rotor having a flat front face and a chamfered or rounded periphery or having a rounded front face; Preferably the rotor has a peripheral terminal collar at a first end of the rotor that extends across the cylindrical rotor portion; the collar, spaced from the stator, preferably extends across the first end cross-sectional opening of the stator and / or across the cylindrical surface of the stator; Accordingly, the collar has a spacing that extends across an annular gap formed between the rotor and the stator. A rotor, - a first end section of the housing is covered by an integral bearing block including a first bearing for a shaft, the first end section of the shaft being connected to the shaft drive and an opposite second end of the shaft being connected to the rotor, the bearing block being connected to the housing by a form fit and / or a friction fit, the bearing block preferably having a first stop surface abutting the lid and an opposite second stop surface abutting the first end of the housing, a shaft having a first end to which a magnetic shaft drive is fixed and an opposite second end covered by and fixed to a rotor, The shaft extends into a first bearing of the bearing block, the first bearing being disposed on the shaft at an axial portion disposed between the magnetic shaft drive and the rotor, and preferably the first end of the shaft is a free end and disposed in a recess in the lid, the free end of the shaft and the recess in the lid forming a second bearing which is a friction bearing; A shaft and - the shaft, the magnetic shaft drive, the first bearing and the second bearing from which the shaft extends, the rotor, the stator, preferably the stator extension, and / or the detector are arranged coaxially around the longitudinal axis of the housing; - preferably a lid covering the first end of the housing, preferably connected to the bearing block and / or the housing by form-fit and / or friction-fit, preferably having a recess arranged coaxially with the housing, preferably having a cylindrical side wall that clamps onto the cylindrical portion of the bearing block and / or clamps onto the cylindrical portion of the first end of the housing, The lid and The device may comprise or consist essentially of:

[0012] A stop surface formed between the first and second portions of at least one of the ribs, preferably at least three of the ribs, with the first portion extending further relative to the central longitudinal axis than the second portion, is positioned adjacent to the second end of the stator so that the stop surface of at least one rib, preferably each rib, contacts the second end of the stator. During assembly of the reactor vessel, the stator moves along the longitudinal axis from the first end of the housing toward the second end of the housing until it abuts against the stop surface. The second portions of at least three of the ribs, each extending the same radial distance from the central longitudinal axis, form at least three contact surfaces for clamping the stator therebetween, such that the stator is held by a friction and / or form fit such that its inner cylindrical wall is rotatable and preferably its outer cylindrical wall is coaxial with the central longitudinal axis, i.e., rotationally symmetrical.

[0013] The first portions of the ribs preferably extend the same distance from the central longitudinal axis, leaving a free volume between them that is assembled to allow liquid to flow through. This free volume is connected to a channel formed between the outer surface of the stator and the housing, and is connected to an opening at the second end of the stator. Because there is no relative movement between the stator and the housing in the gap that forms the channel for liquid, shear forces in this channel are significantly lower—essentially no significant shear forces, for example, compared to the shear forces generated between the rotor and the stator. As a result, in the channel between the stator and the housing, liquid can return to the gap between the rotor and the stator without being exposed to any relevant shear forces. For example, with a stator outer radius of 2.8 mm and a channel width of 1 mm, and a rotor radius of 1.5 mm and a rotor-to-stator distance of 0.3 mm, the shear force generated in the channel is less than 1% of the shear force generated between the rotor and the stator at 3000 rpm. Preferably, the spacing of the stator from the housing is at least as large as the annular gap between the rotor and stator, preferably at least 1.5 times larger, for example up to 3 times larger, to generate maximum shear forces between the rotor and stator and avoid significant shear forces as the liquid flows through the gap between the stator and the housing.

[0014] The first end cross section of the housing spans the entire cross section of the first end of the housing. The bearing block can be connected to the first end of the housing by being clamped to the circumferential surface of the housing, preferably to the inner surface of the housing, by a form fit, preferably a friction fit.

[0015] Typically, the inner cross section of the housing, excluding the rib, is cylindrical, e.g., having the same diameter along the portion of the longitudinal axis along which the first and second rib portions extend, and having the same or a larger diameter along the portion of the longitudinal axis extending from the second rib portion to the first end cross section of the housing. Generally, it is preferred that the second rib portion extends only over the detection portion. At least one detection window, preferably at least two detection windows, are each formed by an optically transparent, flat, constant-thickness wall of the detection portion.

[0016] Preferably, the housing includes a bottom wall and ribs, and the at least one detection window is integrally formed, for example, from an optically transparent synthetic resin, preferably by injection molding. The stator, preferably manufactured by injection molding, is disposed between the second portions of the ribs in a friction and / or form fit, with the second end of the stator abutting against a stop surface formed by the first portion of at least one of the ribs.

[0017] The magnetic shaft drive is disposed on the shaft and is fixed, for example, by a form fit and / or a friction fit, spaced apart from the bearing block, preferably from the first end of the shaft. The magnetic shaft drive may comprise or consist of a shaft extending, for example, as a cylindrical bore, within a friction bearing disposed in the bearing block. The shaft drive is connected to the first end of the shaft, and optionally the shaft protrudes above the shaft drive, with the protruding end of the shaft adapted to be placed within a recess in the lid, which forms a bearing for the first end of the shaft. The shaft drive is disposed between the first end of the shaft and the bearing block, and preferably the lid covers the first end of the shaft and the shaft drive and is connected, for example, by a friction fit and / or a form fit with a portion of the bearing block, for example, the lid can be circumferentially fastened to the bearing block. The rotor is connected, for example, by a friction fit and / or a form fit, to a second end of the shaft opposite the first end of the shaft. Preferably, a circumferential portion of the bearing block opposite the circumferential portion of the bearing block clamped by the lid is adapted to clamp into the first end cross-sectional opening of the housing, for example by a friction fit and a form fit.

[0018] Preferably, the reaction vessel comprises: a housing in which the stator is disposed against the stop surface and is held between the first portion and the second portion of the ribs by a friction fit and / or a form fit; a second part, a bearing block for holding the shaft in a bearing, the bearing block having a shaft drive disposed at a first end of the shaft; a rotor connected to the opposing second end of the shaft, e.g., by a friction fit; a lid connected to the bearing block, the lid covering the shaft drive and preferably clamped to the circumferential surface of the bearing block by a friction fit; or provided as a kit of parts consisting of these, Preferably, the first end of the shaft extends into a central recess in the lid, and the free end of the shaft and the recess in the lid form a second bearing; The circumferential portion of the bearing block is assembled for a friction fit, e.g., a clamping connection, into the first end cross-section opening of the housing, in particular for a friction fit of the portion of the bearing block inside the housing at the first end cross-section of the housing.

[0019] Generally, the rotor, rotor peripheral collar, stator, any extension pipes, and first and optional second bearings and magnetic shaft drive are coaxial with the shaft, and preferably all of the vessel components are coaxial with a common longitudinal axis, for example the longitudinal axis of the shaft.

[0020] In a detection section having two windows, the windows are preferably arranged at 90°, more preferably parallel to each other on opposite sides of the housing wall.

[0021] Typically, the stator is spaced apart from the housing, the spacing of the stator from the housing being provided by the second portions of the ribs, the spacing forming a channel for liquid flow from one end portion of the stator, e.g., from a first end cross-section of the stator, to an end opposite the first end cross-section, e.g., to an opening in the second end cross-section. Typically, the stator may have a tapered, e.g., funnel-shaped, portion at its second end that includes the second end opening.

[0022] Preferably, the cross-sectional area of ​​the extension pipe is at least as large as the cross-sectional area of ​​the annular gap between the rotor and the stator. Preferably, the extension pipe is circular in each of its cross-sections or heights.

[0023] Preferably, the rotor is spaced from the bearing block so that the shaft is not covered due to the spacing. Providing a spacing between the rotor and bearing block reduces contamination of the bearing block by liquids contacting the rotor.

[0024] Preferably, the cross section of the housing between the height of the lid or the height of the first bearing, for example, located in the lid, and the height of the rotor, has a diameter smaller than the outer or inner diameter of the stator and / or the diameter of the rotor, particularly the diameter of the rotor's peripheral collar. Such a smaller diameter portion is also referred to as the collar portion of the housing. Preferably, the collar portion covers the portion of the housing between the lid or the first bearing, for example, located in the lid, and the rotor, while the shaft is not covered by either the first bearing or the rotor. In a process using a container, preferably, liquid is poured into the housing up to the smallest cross section of the collar portion, preferably to a level that completely fills the annular gap between the rotor and the stator.

[0025] The housing preferably has a recess along its inner wall, preferably of cylindrical cross section, at its first end for receiving the cylindrical portion of the bearing block and for abutting the second stop surface of the bearing block, preferably with a friction fit, the friction fit optionally including grooves and ridges on the outside of the cylindrical side wall of the bearing block mating with grooves and ridges on the inner surface of the recess at the housing first end. Preferably, the bearing block is fixed to the lid, e.g., clamped, e.g., the bearing block is clamped into the recess formed by the cylindrical side wall of the lid. The inner wall of the lid on which part of the bearing block is located, e.g., the part abutting the first stop surface of the bearing block, may have grooves and ridges assembled to mate with grooves and ridges on the outer surface of the bearing block.

[0026] The shaft is preferably a cylindrical stainless steel rod, although alternatively the shaft can be a cylindrical rod of high performance plastic.

[0027] The first bearing disposed in the bearing block can be a bore, which optionally provides a friction bearing for the shaft with only radial guidance and / or without axial guidance. The magnetic shaft drive preferably comprises or consists of a holder containing a magnet, ferromagnetic pin, or ferrimagnetic pin, which is fixed to the shaft, for example by clamping. The holder preferably has a recess for holding the magnet, ferromagnetic pin, or ferrimagnetic pin, preferably one or two pairs of magnets, ferromagnetic pins, or ferrimagnetic pins. Along the shaft, the magnetic shaft drive is preferably arranged between a lid, which has a recess forming a second bearing for the first end of the shaft, and the first bearing of the bearing block is, for example, clamped to both the housing and the lid, so that the magnetic shaft drive holds the shaft between the bearing block and the lid, even if the first bearing is a bore that allows the shaft to slide axially. To prevent axial movement of the shaft in the first bearing, the magnetic shaft drive can be secured adjacent to the bearing block, e.g., where the magnetic shaft drive abuts the bearing block, and the first end of the shaft extends into the second bearing, with the front surface of the shaft abutting the recess in the lid with sufficient clearance to allow rotation of the shaft. Preferably, the magnet is a neodymium magnet, e.g., a cylindrical magnet disposed within the bore of the holder. Alternatively, other high-performance magnets can be used, including high-performance composite magnets fabricated by injection molding.

[0028] Preferably, the shaft is a cylindrical metal rod, the magnet of the magnetic shaft drive is a neodymium magnet, and all other components of the vessel are preferably optically transparent synthetic resins, such as polystyrene, polyethylene, polylactic acid, polyethylene terephthalate, polycarbonate, or acrylic nitrile-butadiene-styrene, or nylon, or polypropylene.

[0029] For example, when attaching components of a reaction vessel, the components of synthetic resin, which are fixed to one another by clamping their sliding surfaces against one another, preferably have a groove perpendicular to their common longitudinal axis, for example perpendicular to the longitudinal axis of the shaft, which groove is manufactured by additive manufacturing, for example by 3D printing, preferably liquid resin 3D printing, for example using high-precision fused molding (FDM) technology, preferably by injection molding.

[0030] The magnetic shaft drives can be driven by magnetic drives having correspondingly arranged magnets, preferably the same number of magnet pairs as there are shaft drives.

[0031] Preferably, the bearing block is connected to the lid, preferably inside a recess formed by the cylindrical side wall of the lid, for example by clamping, and the bearing block is assembled to connect to the first end of the housing by clamping, which allows the container to be provided as a separate element that is easy to connect by clamping without the need for additional fixing means, for example without additional adhesives, seals or mechanical fastening devices.

[0032] For the analytical process, the container is: - a housing with a detection portion having at least one, preferably at least two, optically transparent windows at a second end of the housing, the housing including as a separate element a stator arranged by friction and / or form fit between second portions of ribs and abutting against a stop surface formed by a first portion of at least one rib, the ribs being integrally formed with the housing, the housing having at its first end a recess assembled to receive a bearing block; As a separate part - a lid connected to a bearing block including a shaft having a rotor fixed to a second end of the shaft and a magnetic shaft drive fixed to a first end of the shaft, the bearing block being assembled so as to be attached to the first end of the housing with a friction fit and / or a form fit, The lid and can be set in combination with The analysis process includes the following steps: - injecting the liquid to be analyzed into the housing and then placing a bearing block at a first end of the housing, the rotor being coaxially positioned within the stator; - applying a rotating magnetic field to the magnetic shaft drive and optically analyzing the liquid through a detection window; It has.

[0033] The reaction vessel has the advantage that it is constructed so that, when the rotor is rotated, a shear force is exerted between the rotor and the stator, providing a pumping action on the liquid, resulting in homogeneous processing of the entire liquid in the vessel, and resulting in a liquid representative of the entire liquid being present in the detection portion of the housing. The peripheral collar extending across the cylindrical rotor portion, when rotated, can generate sufficient centrifugal force on the liquid to exert a pumping action on the liquid, causing the liquid to move through the gap between the rotor and the stator, exit the first end of the stator, circulate through the channel formed by the gap between the housing and the stator, then pass through the detection portion and return to the opening provided in the second end of the stator.

[0034] Since pumping action is provided solely by rotation of the rotor, the vessel drive element, e.g., the rotating element, preferably comprises a rotor disposed at the second end of the shaft and a shaft drive disposed at the first end of the shaft. Furthermore, the vessel has the advantage of exerting significant shear forces on the liquid substantially only between the rotor and the stator, and thus shear forces can be controlled by controlling the rotational speed of the rotor alone, while the vessel is constructed to avoid generating associated shear forces outside a predetermined ring-shaped gap between the rotor and the stator. Thus, the vessel is constructed to exert maximum shear forces on the liquid only in the ring-shaped gap between the rotor and the stator. The vessel is constructed to circulate the entire volume of liquid through the ring-shaped gap between the rotor and the stator when the rotor is rotated, and only in the ring-shaped gap between the rotor and the stator is the maximum shear force exerted to convert the native-structure prion protein to its aggregated state, while the flow of liquid through the vessel outside this ring-shaped gap does not provide sufficient shear forces to significantly affect the conversion of the native-structure prion protein to its aggregated state.

[0035] The ring-shaped gap between the rotor and stator is preferably 5% to 100% of the rotor radius and / or has a radial width in the range of 0.1 mm to 1.0 mm. The shear stress is proportional to the rotor rotation frequency, the rotor radius, and the viscosity of the liquid, and the shear stress is inversely proportional to the gap width between the rotor and stator.

[0036] Optionally, at least two containers are arranged in parallel and connected to one another, and their housings are manufactured as a single unit, for example by injection molding, wherein the at least two containers connected in parallel to one another are arranged such that their optically transparent windows are arranged in a common plane, for example, the windows are arranged in a plane parallel to the line of rows in which the containers are arranged in the array.

[0037] The present invention further provides an analytical method in which a liquid sample is injected into a container, preferably by injecting the sample into a housing and / or a stator, then placing a rotor in the stator, preferably the rotor is mounted on a shaft extending on a first bearing fixed to the lid, and placing the lid on a first end of the housing, rotating a magnetic shaft drive to rotate the rotor, optically detecting the sample in a detection unit of the housing, and preferably transmitting the detection results or a medical indicator derived from the detection results to the sample provider. The sample provider may be a medical research institution, a physician, or a patient from whom the sample was collected. Furthermore, the analytical method can be used to determine the effectiveness of a compound in inhibiting or reversing the formation of aggregated prion protein by adding the compound to the sample or an aliquot of the sample and comparing the rate of formation of aggregated prion protein during the process. Thus, this processing method can be used to analyze samples from patients by adding a compound suspected of being effective against the formation of aggregated prion protein to the sample from the specific patient to detect the efficacy of the compound in at least delaying the formation of aggregated prion protein in the donor's sample. The effectiveness of a compound in delaying the formation of aggregated prion protein includes preventing, inhibiting, suppressing, and / or reversing the formation of aggregated prion protein. Here, the processing method can be used to select compounds for their effectiveness in delaying, e.g., inhibiting, suppressing, preventing, or reversing, the formation of aggregated prion protein for a particular patient sample. Generally, the sample can be a liquid or solid biological material, such as a solution, serum, or tissue, obtained from a patient. Optionally, aliquots from the mixture containing the sample and native prion protein can be removed from the container at different times during processing in the container, e.g., by stopping the rotor, removing an aliquot, optionally rotating the rotor further, stopping it again, and removing another aliquot. It is generally preferred to freeze and store the aliquots removed from the container, e.g., for further analysis. The patient can be a human patient, or an animal or tissue culture, particularly for research purposes.The processing method is an in vitro process or assay and can be used, for example, as a translational assay system during drug discovery.

[0038] The process using the device of the present invention can also be used to screen and select compounds for their activity and effectiveness in inhibiting, suppressing or reversing the formation of aggregated prion protein, the process comprising: injecting a liquid sample comprising native prion protein and / or aggregated prion protein; adding at least one compound to be screened into the housing and placing the rotor inside the stator; rotating the magnetic shaft drive to rotate the rotor; optically detecting the sample in a detection portion of the housing; detecting a compound having an activity of delaying the formation of aggregated prion protein; Equipped with.

[0039] The invention will now be explained in more detail with reference to examples and figures. [Brief explanation of the drawings]

[0040] [Figure 1] FIG. 1 shows a cross-sectional view of one embodiment of a reaction vessel. [Figure 2] FIG. 2 shows a view of section BB of FIG. [Figure 3] FIG. 3 shows a view of section AA of FIG. [Figure 4] FIG. 4 shows an exploded view of the reaction vessel. [Figure 5] FIG. 5 shows an exploded view of the elements directly connected to the bearing block. DETAILED DESCRIPTION OF THE INVENTION

[0041] Generally, functionally identical elements are designated with the same reference numerals.

[0042] 1 shows a housing 1 having a first end 2 and an opposing second end 3, the second end 3 being closed by a bottom wall 4. A detection unit 5 is disposed at the second end 3 of the housing 1, the detection unit 5 having at least one, and preferably two, opposing optically transparent windows 6. Optionally, the bottom wall 4 has a bottom window 4a of optically transparent material, optionally a central portion of which is surrounded by an opaque bottom wall 4 forming a bottom opening 4b.

[0043] Disposed within housing 1 is a stator 10 extending from a first open end cross section 11 of the stator to an opening 12 at a second end 12a opposite first open end cross section 11, stator 10 having a cylindrical inner surface 15. At second end opening 12, the stator has a distal end 13 tapering to opening 12, which opens into sensing portion 5 of housing 1.

[0044] The stator 10 is held in the housing by a friction fit between the second portions 14b of the ribs 14. The stator 10 extends along the surfaces of the second portions 14b of the ribs 14. The outer surface of the stator 10 is cylindrical, and the second portions 14b of the ribs 14 are held rotationally symmetrically between the second portions 14b so that they extend the same distance from the central longitudinal axis 7 and from the inner surface of the housing 1. During assembly, the stator 10 can move along the central longitudinal axis 7 between the second portions 14b until it abuts the stop surfaces 8 formed by the first portions 14a of the ribs 14.

[0045] The rotor 20 is coaxially disposed inside the stator 10, and the spacing between the rotor 20 and the cylindrical inner surface of the stator 11 forms an annular gap where rotation of the rotor 20 applies shear forces to the liquid. The rotor 20 is disposed within the stator 10 as a cylindrical portion 21, and at its first end 22, the cylindrical portion 21 has a terminal peripheral collar 23 that extends across the radius of the cylindrical rotor portion 21. The collar 23 of the rotor according to the preferred embodiment extends across the annular gap formed between the rotor 20 and the stator 10.

[0046] The opposing second end 24 of the rotor has a flat front surface 25 with a circumferential bevel 26 .

[0047] The rotor 20 is disposed at a second end 32 of the shaft 30. A first end 31 of the shaft 30, opposite the second end 32, carries a magnetic shaft drive 33 including at least two magnets 34. The magnetic shaft drive 33 is fixed to the shaft 30, which extends in a first bearing 35 in a bearing block 39, the shaft 30 being optionally axially displaceable. In the embodiment shown, the magnetic shaft drive 33 has a flat front face 36 that can extend with friction on a flat front face 37 adjacent the bearing block 39, which limits the axial movement of the shaft 30.

[0048] Bearing block 39 is fastened to the inside of cylindrical portion 41 of lid 40, which abuts a first stop surface 43 of bearing block 39. Lid 40 extends over bearing block 39 and encloses shaft drive 33. Bearing block 39 has a second stop surface 44 that abuts a front surface at first end 2 of housing 1.

[0049] The shaft 30 extends into a first bearing 35 in a bearing block 39, and a first end 31 of the shaft 30 is attached by clamping to a magnetic shaft drive 33. In a preferred embodiment, the first end 31 of the shaft 30 terminates in a convex front surface 38 that extends into a corresponding recess 42 in the lid, the front surface 38 of the shaft 30 forming a second bearing 50 with a corresponding recess 42 in the lid 40.

[0050] In the illustrated embodiment, all elements are arranged coaxially with the central longitudinal axis 7 of the shaft 30 .

[0051] 2 shows cross section BB of second portions 14b of ribs 14, between which second portions 14b stator 10, with rotor 20 spaced apart and coaxially arranged, is clamped by a friction fit. Between the outer surface of stator 10 and housing 1, a channel 17 restricted by ribs 14 opens, which is in fluid communication with first open end cross section of stator 11, runs along the outside of stator 10, and is in fluid communication with sensing element 5 and opening 12 at second end 12a of stator 10. Second portions 14b of ribs 14 define a central opening 16 that is assembled to receive stator 10 in a friction fit.

[0052] 3 shows cross section AA of stop surface 8 formed by first portion 14a of rib 14, which extends further toward central longitudinal axis 7 than second portion 14b. When stator 10 is placed with its second end 12a abutting against stop surface 8, channel 17 is formed between housing 1, the outer surface of stator 10, and first portion 14a of rib 14.

[0053] 4 shows the pre-assembled elements of the vessel, which preferably comprise or consist of a rotor 20 attached to the second end 32 of a shaft 30, which extends in a first bearing of a bearing block 39, to which a lid 40 is connected by a friction fit. The lid 40 covers a magnetic shaft drive 33, which is arranged adjacent to the first end 31 of the shaft 30. Another pre-assembled element for producing the vessel is a housing 1, which is provided inside the housing 1 with a stator 10 arranged between the second portions 14b of the ribs 14.

[0054] The exploded view, Figure 5, shows the single elements, which are preferably attached to one another by a friction fit, namely the shaft 30, the rotor 20 for placement on the second end 32 of the shaft 30, the bearing block 39 having drillings forming a first bearing 35 for the shaft 30, the magnetic shaft drive 33 for placement on the first end 31 of the shaft 30, and the lid 40 which clamps the bearing block 39 and abuts against the first stop surface 43 of the bearing block 39.

[0055] Example: analytical process in which the effect of shear forces on a sample is detected by optical detection As a typical sample, recombinant aggregated prion protein derived from a shear-force-induced reaction of recombinant native prion protein was mixed with 1.5 mg / ml native human α-synuclein in PBS containing 1% Triton X-100 at a dilution of 1 / 100,000, along with a postmortem brain homogenate sample from a synucleinopathic patient (positive control). The buffer composition and procedural details were similar to those previously disclosed in U.S. Patent Nos. 5,999,112 and 5,999,123. The fluorescent substance used to detect aggregated prion protein was thioflavin T. As a negative control, the same reaction composition was used without the addition of recombinant aggregated prion protein.

[0056] The vessel generally corresponds to Figure 1. The gap width between the stator and rotor is 0.3 mm, and the rotor diameter is 3 mm. The treatment frequency used was 400 revolutions per second (400 Hz), corresponding to a shear rate of 12,570 revolutions per second and a shear stress of 11.2 Pascals, and the reaction temperature was set at 30 °C. Overall, the reaction was followed for 15 hours, corresponding to 180 cycles of treatment and rest. In each cycle, the treatment was applied for 3 seconds, followed by a 297-second rest phase. To detect the formation of aggregated prion protein, the fluorescence signal of Thioflavin T was accumulated during the rest phase of each cycle. Data traces for 15 replicates were recorded for each of the positive and negative controls.

[0057] In the positive control, aggregated prion protein formed after approximately 4 hours (±0.5 hours). In the negative control, aggregated prion protein began to form after approximately 14 hours in some replicas, but most replicas showed no signs of aggregated prion formation during the observation period. The measurement results are shown in Figure 4, along with the positive control (left) and negative control (right). The individual measurements overlap because aggregate formation occurred in a reproducible manner.

[0058] Typically, detection can be measured by changes in fluorescence of a fluorescent dye added to the mixture through a detection port in a container. The dye is specific to the aggregated prion protein. Typical dyes include thiophene-based amyloid ligands such as thioflavin T, thioflavin S, Congo Red, luminescent conjugated polythiophenes (LCPs), polythiophene acetic acid (PTAA), and luminescent conjugated oligothiophenes (LCOs), Pittsburgh compound B, aminonaphthalene 2-cyanoacrylate (ANCA) probes, PEGylated phenylbenzoxazole derivatives, pinacyanol, chrysamine G, and dyes containing at least one of the following scaffolds: chalcone, flavone, aurone, stilbene, diphenyl-1,2,4-oxadiazole, diphenyl-1,3,4-oxadiazole, benzothiazole, benzoxazole, benzofuran, imidazoviridine, benzimidazole, quinoline, and naphthalene.

[0059] Alternatively, native conformation prion protein can be labeled with a fluorescent dye by, for example, attaching the fluorescent dye directly to the native conformation prion protein or via an intermediate spacer, such as a fluorescent derivative containing a reactive chemical group, such as an isothiocyanate (which reacts with primary amines, e.g., lysine), a succinylimide ester (which reacts with amino groups to form an amide bond, e.g., the N-terminal amino acid), or a maleimide (which reacts with free sulfhydryl groups, e.g., cysteine). Such fluorescent derivatives include cyanines, fluoresceins, rhodamines, Alexa fluorescers, Dylite fluorescers, ATTO-Tec dyes, BODIPY dyes, SETA dyes, SeTau dyes, and DYOMICS dyes. [Explanation of symbols]

[0060] 1. Housing 2 first housing end 3 Second housing end 4 Bottom wall 4a Bottom window 4b Bottom opening 5. Detection unit 6 Window section 7 central longitudinal axis 8 Stop surface 10 Stator 11 first stator end 12 Opening in second end cross section of stator 12a Second end of stator 13 End portion tapering towards opening 14 Ribs 14a First part of the rib 14b Second part of the rib 15 Cylindrical inner surface of stator 16 Central opening 17 channels 20 rotors 21 Cylindrical part of rotor 22 first end of rotor 23 Rotor color 24 second end of rotor 25 Front of rotor 26 Slope 30 shaft 31 first edge of shaft 32 second edge of shaft 33 Magnetic Shaft Drive 34 Magnet 35 First bearing 36 Front of magnetic shaft drive 37 Front of bearing block 38 Front of shaft 39 Bearing block 40 Lid 41 Cylindrical part of the lid 42 Recessed portion inside the lid 43 First stop surface of bearing block 44 Second stop surface of bearing block 50 Second bearing

Claims

1. A reaction vessel used for analyzing a sample, the vessel comprising: a housing (1) extending along a central longitudinal axis (7), said housing (1) having a first end (2) for connection to a lid (40) and an opposing second end (3) closed by a bottom wall (4), said housing (1) having a detector (5) arranged at said second end (3), said detector (5) comprising at least one window portion (6) in a wall of said housing, said at least one window portion (6) being optically transparent; - the housing (1) comprises at least three ribs (14) extending along the housing wall to the inside of the housing (1) and projecting towards the central longitudinal axis (7); The rib (14) adjacent to the first end (2) of the housing has a first portion (14a) and a second portion (14b) adjacent to the first portion; the second portion (14b) of the rib (14) extends towards the longitudinal axis (7); a first portion (14a) of at least one of said ribs (14) extending further toward the longitudinal axis than a second portion (14b) thereof; The rib comprises at least one rib (14) having a stop surface (8) formed between the first portion (14a) and the second portion (14b), Housing and a stator (10) disposed within a housing (1), the stator (10) having a cylindrical inner surface (15) for receiving a rotor (20) at a fixed distance, the cylindrical inner surface (15) having a first open end cross-section (11) and an opening (12) at a second end (12a) opposite the first open end cross-section, the outer surface of the stator (10) being disposed at a distance from the housing (1); The form-fitted and / or friction-fitted stator (10) is arranged such that a first end (11) of the stator abuts against a stop surface (8), the stator (10) is disposed between the second portions (14b) of the at least three ribs (14) in a friction fit and / or a form fit; The ribs (14), the stator (10) and the wall of the housing (1) define a channel (17) outside the stator (10), the channel (10) having an open end cross section at both ends thereof. a stator; a rotor (20) having a cylindrical portion disposed within the stator (10) at a distance that defines a ring-shaped gap of constant radius, the rotor (20) having a second end (24) that covers the second end (32) of the shaft (30); a rotor; a cross section of the first end (2) of the housing (1) is covered by a one-piece bearing block (39) including a first bearing (35) with a shaft (30), a portion of the first end (31) of the shaft (30) being connected to a shaft drive (33) and an opposite second end (32) of the shaft (30) being connected to the rotor (20), the bearing block (39) being connected to the housing (1) by a form-fit and / or friction-fit; - said shaft (30) having a first end (31) to which a shaft drive (33) is fixed and an opposite second end (32) covered by and fixed to said rotor (20); the shaft (30) extends into a first bearing (35) of the bearing block (39), the first end (31) of the shaft (30) is a free end and is disposed in a recess (42) of the lid (40), and the shaft free end and the recess of the lid (40) form a friction bearing; - said shaft (30), said shaft drive (33), said first bearing (35), said rotor (20) and said stator (10) are arranged coaxially around one longitudinal axis (7); - the lid is connected to the bearing block (39) and / or the housing (1) by form-fit and / or friction-fit; container.

2. 2. The container according to claim 1, wherein the housing (1) and the ribs (14) are integral and the stator (10) is held between the second portions (14b) of the ribs (14) by a friction fit.

3. 3. A container according to claim 1 or 2, characterized in that the first portion (14a) and the second portion (14b) of each rib (14) are arranged parallel to a central longitudinal axis (7).

4. 4. A vessel according to any one of claims 1 to 3, characterized in that the outer circumferential surface of the stator (10) is cylindrical and the end cross-section of the stator (10) consists of a ring-shaped surface perpendicular to the central longitudinal axis (7).

5. 5. A vessel according to any one of claims 1 to 4, characterized in that the rotor (20) has, at its first end (22), a terminal peripheral collar (23) extending across a radius of the cylindrical portion of the rotor (20), the collar (23) being spaced apart from the stator (10) and extending across a cross-sectional opening of the first end (11) of the stator (10).

6. 6. A container according to any one of claims 1 to 5, characterized in that the bearing block (39) has a first stop surface (43) abutting against the lid (40) and an opposing second stop surface (44) abutting against the first end (2) of the housing (1).

7. 7. The container according to claim 1, wherein the bearing block (39) is arranged on the shaft (30) in an axial portion arranged between the shaft drive (33) and the rotor (20).

8. 8. The container according to any one of claims 1 to 7, characterized in that the lid (40) has a cylindrical portion (41) of a side wall forming a recess arranged coaxially with the housing (1), the cylindrical side wall being fastened to the cylindrical portion (7) of the first end (2) of the housing (1).

9. 9. A container according to any one of claims 1 to 8, characterized in that the collar (23) extends across the annular gap formed between the rotor (20) and the stator (10).

10. 10. The container according to any one of claims 1 to 9, characterized in that the first end (31) of the shaft (30) is a free end that is arranged in a recess (42) of the lid (40), and the free end (31) of the shaft and the recess (42) of the lid (40) form a second bearing (50), which is a friction bearing.

11. The container is provided as an element including the housing (1), the stator (10) disposed inside the housing (1), and the detection unit (5), the lid (40) is provided as a separate element connected to the bearing block (39) containing the first bearing (35); In the first bearing (35), the shaft (30) extends to a first end (31) attached to a magnetic shaft drive (33); The magnetic shaft drive (33) is disposed between the first bearing (35) and the lid (40); The first end (31) of the shaft (30) is a free end that is disposed in a recess (42) of the lid (40); The free end (31) of the shaft and the recess (42) of the lid (40) form a second bearing (50); The rotor (20) is attached to the second end (32) of the shaft (30); Container according to any one of claims 1 to 10, characterized in that

12. Container according to claim 11, characterized in that the housing (1) has, at its first end (2), a cylindrical portion adapted to receive the bearing block (39) with a clamping fit.

13. A process for analyzing a sample from a patient to detect the presence of aggregated prion protein using a container according to any one of claims 1 to 12, comprising the steps of: The process is as follows: injecting the sample and native prion protein into the housing; placing the rotor inside the stator; rotating a magnetic shaft drive to rotate the rotor; optically detecting the sample at the detection portion of the housing; transmitting the detection result or a medical indicator derived from the detection result to the provider of the sample; The process includes:

14. 14. A process for analyzing a sample from a patient according to claim 13, comprising: The process includes adding a compound to the sample from the patient to detect the effectiveness of the compound to slow or inhibit or reverse the formation of aggregated prion protein in the sample from the donor. process.

15. A process for screening a compound for its activity of inhibiting the formation of aggregated prion protein, using the container according to any one of claims 1 to 12, comprising: The process comprises the following steps: Injecting a liquid sample containing native prion protein and / or aggregated prion protein; adding at least one compound to be screened in the housing; placing the rotor within the stator; rotating the magnetic shaft drive to rotate the rotor; optically detecting the sample in a detection portion of the housing to detect a compound having activity to delay, inhibit or reverse the formation of aggregated prion protein; The process includes:

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