Biocompatible composite element and method for manufacturing the same

Abstract

To provide a biocompatible composite element for enabling spectral process control and / or provision of real-time data in a disposable reactor.SOLUTION: A biocompatible composite element 10 for forming a window in a bioreactor is for use as a connection fixture for the bioreactor, and comprises: an outer frame 20 having or composed of a polymeric material for inseparable attachment to a wall of a disposable bioreactor; and an internal component 30 having or composed of a transparent material 32 such as glass, sapphire or glass ceramic. The internal component is housed in an outer frame so that it cannot be separated in a sterile sealed state, preferably in a hermetically sealed state, and a window is formed, to thereby allow spectral process control from outside the bioreactor.SELECTED DRAWING: Figure 1

Description

[Technical Field]

[0001] The present invention relates to spectral process control of manufacturing processes in bioreactors.

[0002] Bioreactors are used to culture microbial, animal, and plant cells, thereby opening up a wide range of applications for biotechnology manufacturing processes. In general, there is a need for further optimization of these processes. In particular, for the production of biopharmaceuticals, there is a growing demand for improved product yields and, therefore, increased profits. Various approaches are available to increase product yields, minimize contamination risks, and reduce costs.

[0003] On the one hand, real-time process control of key parameters can be carried out to optimize process control and therefore yield. Spectroscopy is particularly suitable for this, as it allows measurements to be taken from outside the bioreactor, thus avoiding contamination risks.

[0004] In the case of bioreactors with stainless steel culture vessels, such measurements can be made possible by providing a window into the interior of the bioreactor, for example by fitting flanged connection fittings with an inspection window, which can be formed, for example, as a metal-glass composite, forming, for example, a so-called Glass-To-Metal-Seal (GTMS).

[0005] On the other hand, cost reductions in biotechnological production processes can also be realized by replacing conventional stainless steel bioreactors with disposable or single-use bioreactors. The use of disposable materials in this case, especially pre-sterilized plastics, allows for significant cost savings, depending on the intended use, not only during procurement but also during operation, particularly by avoiding repeated cleaning and sterilization and the associated cleaning validation.

[0006] Even in the case of disposable reactors, yields are highly dependent on measurements of substrate and product concentrations. Nevertheless, in-situ process control by conventional techniques is not yet fully available for single-use applications. The use of glass-to-metal seals (GTMS) is not well-served for single-use applications.

[0007] Therefore, it is common practice in the industry to sample during cultivation, which takes a significant amount of time. This requires sampling, which also carries the risk of contamination, and offline analysis, which is resource-intensive. Despite this effort, the lack of real-time data hinders process control, especially in single-use applications.

[0008] The present invention is therefore based on the problem of being able to provide spectral process control and / or real-time data even in single-use bioreactors. One aspect of this problem is therefore to provide an alternative to the use of glass-to-metal connection fittings, especially for applications where a glass-to-metal seal (GTMS) is not preferred.

[0009] This problem is solved by the subject matter of the independent claims. Advantageous embodiments of the invention are defined in the dependent claims.

[0010] The present invention provides a biocompatible composite element, particularly for use as a connection fitting for a bioreactor or other vessel, preferably for use as a connection fitting for a disposable bioreactor.

[0011] The composite element comprises an outer frame made of or consisting of a polymer material, in particular for attachment to the wall of a bioreactor, preferably for inseparable attachment to the wall of a disposable bioreactor, and an inner component made of or consisting of a transparent material, such as glass, in particular borosilicate glass, quartz, sapphire or glass ceramic. The inner component is accommodated in the outer frame in a sterile, sealed, preferably hermetically sealed manner, and forms a window, by which spectral process control can be achieved, in particular from outside the bioreactor. In this case, the inner component is accommodated in the outer frame in a particularly inseparable, i.e. permanently fixed manner.

[0012] The composite element forms at least a sterile, preferably hermetic seal, especially between the polymer frame and the internal component. In particular, if the internal component consists of or contains glass, the term Glass-To-Polymer-Seal (GTPS) applies. Therefore, the seal is particularly important in material combinations involving polymers. Suitable polymers for the outer frame, especially in the form of homopolymers, copolymers, or terpolymers, include polyethylene (PE), polypropylene (PP), polyamide (PA), polyethylene terephthalate (PET), ethylene vinyl alcohol (EVOH), polyether ether ketone (PEEK), polyaryl ether ketone (PAEK), polysulfone (PSU), and polyphenylene sulfide (PPS).

[0013] In the sense of the present invention, a sterile seal is understood to mean a seal that exhibits minimal sealing action. The sealing action is determined by a leak rate test using helium according to vacuum method A3 of DIN EN 1779. In this method, the composite element according to the invention is subjected to a time-dependent test. A helium leak rate of 6 x 10 after 2 minutes is determined. -4 A sterile seal is said to be achieved when the helium leak rate is less than 10 mbar·l / s for 2 minutes. -8 A good form of sterile sealing is present when the leak rate has a value of less than mbar·l / s. Preferably, these leak rates are also achieved in tests of 4 minutes each.

[0014] A hermetic seal is also preferred. 1.69 x 10 -10 Seals with a helium leak rate of less than mbar·l / s are called hermetic seals.

[0015] Composite elements or glass-to-polymer seals are specifically designed for general industrial usability. In particular, composite elements or glass-to-polymer seals are suitable for disposable / single-use bioreactors. In particular for single-use bioreactors, the polymer outer shell allows for sterile sealing and welding to the bioreactor wall, e.g., polyethylene (PE) to polyethylene (PE). Thus, the polymer material of the outer shell can match the polymer of the bioreactor. However, different materials are also possible. Thus, the composite element can be permanently and securely attached to the bioreactor wall, such that it can no longer be separated from the wall.

[0016] As explained above, the composite element according to the present invention is biocompatible. This means that, in particular, unwanted pathogens cannot adhere to and / or grow on the composite element during cultivation in a bioreactor. The bonding areas between the glass and the polymer and / or between the metal and the polymer are particularly important. Biocompatibility, i.e., the prevention of harmful adhesion and / or growth of pathogens, can be achieved by the absence of edge angles of 90° or more at the mentioned material transitions. In this context, it should be understood that edge angles refer to angles on a microscopic scale that are important for pathogens, and that the composite element can, of course, have elements and / or areas arranged perpendicular to the optical elements. The present invention envisages microscopic edge angles between the glass and the polymer and / or between the metal and the polymer being less than 90°. This is achieved, for example, during production, by wetting the glass and / or metal with the polymer at an edge angle of less than 90°, or by introducing, for example, an arched depression into the polymer material immediately after the material transition from metal to polymer or from glass to polymer (see also the drawings and their description).Preferably, it is also possible to envisage a contact angle or edge angle between the materials of less than 85°.

[0017] Therefore, it is preferable that at least one, preferably many, and very particularly preferably all, edge angles between different materials, on a scale that is important for pathogen growth, be less than 90°, preferably less than 85°. In this way, zones at the transition between different materials (e.g., polymer / glass) that are favorable for pathogen populations and therefore detrimental to sterility are avoided. The contact angle is determined, in particular, by the inherent adhesion properties of the polymer between the polymer and other materials.

[0018] The principle of the present invention is therefore to provide a composite element with an outer frame made of or having a polymer material, which can therefore be used rationally in particular in disposable bioreactors, while surprisingly allowing a sterile, sealed and biocompatible bond to be achieved, thereby providing optical functionality for culturing in such bioreactors.

[0019] Thus, the present invention generally allows for higher product yields and more efficient process development through improved process control, and in particular allows for the sterile and more reliable application of in-situ spectral process control.

[0020] Critical to the development and control of biotechnology manufacturing processes is the real-time measurement of sterilization conditions and key parameters for the development and control of biotechnology manufacturing processes.

[0021] Pre-sterilized single-use products are particularly important in this respect. Sterile culture conditions are essential, so the measurement must not pose a contamination risk. For single-use applications, the sterilizability requirements customary in industry are, in particular, gamma radiation. In biotechnology, and especially in the production of biopharmaceuticals, only approved materials are permitted.

[0022] To meet the stringent requirements of biopharmaceutical manufacturing, the materials of the composite elements can be selected to comply with the following standards: FDA approved materials (ICH Q7A, CFR 211.65(a) - Code of Federal Regulations, USP Class, Animal Origin Free, Bisphenol A Free) EMA (European Medicines Agency) EU GMP Guide Part II certified materials ·Chemical resistance in the field - ASTM D 543-06 Biocompatibility, e.g. relating to testing according to the US Pharmacopoeia or ISO 10993.

[0023] The polymeric enclosure of the composite element according to the invention can be configured in various ways.

[0024] In particular, the outer frame of the composite element may be annular in shape so that it completely surrounds the inner component in cross section, where the cross section of the inner component may be of any shape, in particular circular or annular.

[0025] Furthermore, the outer frame may be tubular and thus have first (e.g., proximal) and second (e.g., distal) tubular ends, particularly such that the outer frame can be attached to the wall of the bioreactor via the first tubular end.

[0026] Furthermore, the housing preferably comprises a flange for attachment to the wall of a bioreactor, in particular a single-use bioreactor. If the housing is tubular, the flange is preferably located between the first and second tube ends, but may also be located at the first tube end. The flange can be inseparably connected, for example welded, to the wall of the bioreactor, preferably in a permanently fixed manner.

[0027] The flange and the outer frame are preferably integrally formed in one piece, in particular made from or using the same polymeric material, in which case the polymeric flange can be welded in a sterile, sealed manner to the wall of the polymeric container, in particular to the outer wall of a single-use bioreactor.

[0028] Furthermore, the housing preferably comprises a thread, in particular configured as an external thread, which thread is preferably arranged at the first tube end.

[0029] The internal components of the composite element may also be formed in various ways.

[0030] In one variant, the internal component is formed in one piece, i.e., in particular as a plate-shaped transparent part, which is preferably permanently or inseparably housed in the outer frame in a sterile and / or hermetically sealed state. The transparent part may be, for example, a glass plate. Here, the glass of the transparent part comprises or consists of, for example, quartz glass or borosilicate glass. However, the transparent part may also be formed, for example, as a sapphire plate.

[0031] Preferably, the transparent component exhibits a transmittance of more than 75%, particularly more than 90%, in the spectral range of 190-5500 nm. More preferably, the transparent component exhibits a transmittance of more than 75%, preferably more than 80%, particularly preferably more than 90%, in the spectral range of 190-2800 nm. Even more preferably, the transparent component exhibits a transmittance of more than 75%, particularly preferably more than 90%, in the spectral range of 190-2700 nm.

[0032] Furthermore, the transparent element is preferably formed integrally in one piece. If the outer frame is tubular, the transparent element is preferably arranged at the second tube end or extends substantially to the second tube end, in which case a tubular projection can be envisaged.

[0033] In another variant, the internal component of the composite element is formed in at least two pieces, namely a transparent part, in particular in plate form, and a connecting part, more in particular in ring or tubular form.

[0034] In this case, the connecting part is connected to both the transparent part and the outer frame. The connection between the connecting part and the transparent part is formed in a sterile sealed state, preferably in a hermetic sealed state, and the connection may be formed in a permanently fixed state or inseparable state. Similarly, the connection between the connecting part and the outer frame is also formed in a sterile sealed state, preferably in a hermetic sealed state, and preferably in a permanently fixed state or inseparable state.

[0035] As mentioned above, the transparent element may also be integrally formed in one piece, in particular. Furthermore, the connecting element is preferably also integrally formed in one piece. The transparent element is preferably arranged at the second tube end, and the connecting element preferably extends to the first tube end.

[0036] Preferably, the connecting part completely surrounds the transparent part in cross section, such that the transparent part is contained within the connecting part in a sterile and / or hermetically sealed state, preferably permanently fixed or inseparably attached, and the connecting part is also contained within the outer frame in a sterile and / or hermetically sealed state, preferably permanently fixed or inseparably attached.

[0037] It is further conceivable that the transparent part is in contact only with the connecting part, in which case the transparent part is only indirectly connected to the outer frame.

[0038] The joining parts preferably comprise or consist of a metal, in particular a stainless steel, for example an austenitic-ferritic duplex stainless steel.

[0039] It is also conceivable that the connecting part first surrounds in cross section a peripheral frame of the transparent part, for example a frame having or consisting of glass, and that this surrounding is carried out in such a way that the transparent part is accommodated in the frame in a sterile sealed state, preferably in a hermetically sealed state, the frame is also accommodated in the connecting part in a sterile sealed state, preferably in a hermetically sealed state, and the connecting part is also accommodated in the outer frame in a sterile sealed state, preferably in a hermetically sealed state.

[0040] The joining element preferably further has a profile for increasing the contact area with the outer frame. For example, the outer surface of the joining element may be provided with grooves. Microstructures, which can be produced, for example, by laser machining, may be provided, particularly with regard to the strength and / or contact angle of the composite material. For example, the surface of the joining element may have a first set of grooves formed by a plurality of adjacently extending grooves, which intersect with a second set of grooves, resulting in multiple or numerous protrusions between the grooves (see also Figures 14 and 15).

[0041] Furthermore, alternatively or in addition, it can be envisaged that the connecting part is in contact with the transparent part at its front side and / or connected to the transparent part at its front side, and that this connection is made so that both the transparent part and the connecting part are housed in the outer frame in a sterile and / or hermetically sealed state, each preferably in a permanently fixed or non-separable manner.

[0042] In particular, in the case of front-side bonding, but also in other cases, the bonding part may comprise or consist of a transparent material, in particular the same material as the transparent part, such as glass, sapphire, ceramic, or glass-ceramic. In other words, the composite element may be formed, for example, as a glass / glass / polymer composite, a sapphire / glass / polymer composite, a sapphire / sapphire / polymer composite, a glass / ceramic / polymer composite, a sapphire / ceramic / polymer composite, a ceramic / ceramic / polymer composite, etc. As will be explained in more detail below, adjacent surfaces may be formed as coherent surfaces.

[0043] Alternatively, the joining component may comprise or be made of an opaque material, preferably a ceramic or metal, in particular an oxidizable metal, such as aluminum. Particularly preferably, the joining component has an oxide layer on one side adjacent to the transparent component. This oxide layer can have a thickness of at least 5 micrometers, preferably at least 10 micrometers. In other words, the joining component may be designed in the form of its oxide at the joining surface. By using or manufacturing the joining component from an oxidizable metal, the joining area can be adapted to the melting point of the transparent component, making laser welding possible. If the joining component is made of aluminum (melting point 660°C) and an oxide layer of, for example, up to 30 μm thick is formed on the joining surface, the melting point of this Al2O3 zone increases to 2050°C. This allows the joining area to be adapted to the melting point of the transparent component, for example, made of sapphire, making laser welding possible. The temperature values ​​indicated refer to the GESTIS materials database.

[0044] Composite elements, particularly in the case of one-piece designs but generally, may have compressive stresses, in particular internal components housed within an outer shell under compressive stress.

[0045] This can be achieved in particular by the composite element being manufactured or being manufacturable as follows: the outer frame is expanded relative to the inner component (or vice versa, the inner component is contracted), then the inner component, in particular the transparent part, is inserted into the outer frame, and then the outer frame is contracted relative to the inner component (or vice versa, the inner component is expanded).

[0046] However, in this particular embodiment, the outer frame may also independently comprise or consist of polyetheretherketone (PEEK).

[0047] The manufacture of a composite element having compressive stress can be carried out, for example, by first preparing an outer frame and an inner component, with the inner component being oversized relative to the outer frame. The outer component (or the outer frame and the inner component) can then be heated, for example, to a temperature of at least 100°C, preferably at least 150°C (e.g., 200°C), causing the outer frame to expand (or expand more than the inner component), and the oversized component of the inner component disappears or preferably decreases. Therefore, when the inner component is fitted into the outer frame, it can also exhibit an oversized component. For example, an oversized component of more than 0.01 mm, preferably more than 0.03 mm (e.g., 0.04 mm), is conceivable. However, since excessive oversized components may no longer be practical, the oversized component is preferably less than 0.2 mm, more preferably less than 0.1 mm. Therefore, it is conceivable to insert the inner component into the outer frame already under compressive stress. In this case, compressive stresses of more than 10 MPa, preferably more than 20 MPa, and particularly preferably more than 30 MPa can be expected. When the external component is subsequently cooled (e.g., to 22°C), the compressive stresses can further increase. For example, compressive stresses of more than 50 MPa, preferably more than 75 MPa, and particularly preferably more than 100 MPa can be expected.

[0048] Regardless of whether the internal component fitted into the outer shell exhibits an oversize or not, it is advantageous if the internal component is ultimately accommodated in the outer shell under a compressive stress of at least 50 MPa, preferably more than 75 MPa, particularly preferably more than 100 MPa, when the composite element is still present under normal conditions, especially at room temperature. Often, this compressive stress is not the same at every point. In particular, when the internal component is arranged at the tube end of the tubular outer shell, the compressive stress generally decreases towards the outer end (critical end). However, it is preferably envisaged that the minimum compressive stress occurring (at the critical end) still exceeds 1 MPa, preferably more than 5 MPa, particularly preferably more than 10 MPa (e.g., 11 MPa).

[0049] In particular in the case of two-piece designs, but generally also, it is conceivable that the internal components are housed in a stress-neutral manner within the outer housing.

[0050] This can be achieved, for example, by the composite element being manufactured or being manufacturable in such a way that the outer frame is attached to the inner component, in particular the connecting part and / or the transparent part, from the outside, for example by supplying a liquid polymer material from the outside to the inner component and then curing it.

[0051] However, in this particular embodiment, the outer frame may also comprise or consist of polyethylene (PE) regardless.

[0052] When the transparent part is housed in the connecting part, it can optionally be assumed that the transparent part is under compressive stress, which is the case in particular for connecting parts made of metal, but also in general.

[0053] A transparent part housed in a joining part under compressive stress can be achieved in particular by the internal components being manufactured or being manufacturable in such a way that the joining part is expanded relative to the transparent part (or vice versa), then the transparent part is inserted into the joining part, and then the joining part is contracted again relative to the transparent part (or vice versa).

[0054] If the transparent part is connected to the connecting part at its front side, and in general, it can be assumed that the transparent part is connected to the connecting part in a stress-neutral manner, this is particularly true for connecting parts made from or with the same material as the transparent part, but also in general.

[0055] A stress-neutral connection can be achieved in particular by the internal component being manufactured or being manufacturable in such a way that the connecting part is pressed against the transparent part at the front side and then the connecting part is firmly connected, preferably to the transparent part, in particular by laser welding.

[0056] The above method sequence is explained again in detail below.

[0057] Thus, the biocompatible composite element can be produced, for example, by the following method steps, in particular in this order: a) providing an outer frame comprising or consisting of a polymer material, in particular polyetheretherketone (PEEK), and an inner component comprising or consisting of a transparent part, in particular a transparent part comprising or consisting of glass, sapphire or glass ceramic, b) causing a relative expansion of the outer frame relative to the internal component, said expansion being achieved in particular by heating the outer frame or preferably by heating the outer frame and the internal component together; c) inserting the internal component, in particular the transparent part, into the outer frame inflated relative to the internal component; d) causing relative shrinkage of the outer shell relative to the internal component, in particular by cooling the outer shell, or preferably by cooling the outer shell and the internal component together, so that the outer shell accommodates the internal component in a hermetically sealed state.

[0058] In other words, in step b) and correspondingly in step d), the outer frame (or the outer frame and the internal components) can be subjected to positive or negative thermal expansion, which is carried out in such a way that a temperature-dependent thermal expansion difference is achieved between the outer frame and the internal components, with positive thermal expansion (expansion) being carried out in particular in step b) and negative thermal expansion (contraction) being carried out in step d).

[0059] The biocompatible composite element can furthermore be produced, for example, by the following method steps, in particular in this order: a) providing an internal component and a liquid polymer material for forming an outer frame; b) supplying a liquid polymer material from the outside to the internal components, in particular to the bonding part and / or the transparent part; c) hardening the liquid polymer material to form an outer shell that encases the internal components in a sterile, sealed, preferably hermetic, condition;

[0060] To prepare the internal component in step a), the following method steps are further carried out, in particular in this order, before the above method sequence: aa) providing a transparent part and a joining part, in particular a joining part made of metal; bb) inducing a relative expansion of the joining part with respect to the transparent part, said expansion being achieved in particular by heating the joining part or preferably by heating the joining part and the transparent part together; cc) inserting the transparent part into the joining part expanded relative to the transparent part; dd) inducing a relative shrinkage of the joining part with respect to the transparent part, in particular by cooling the joining part or preferably by cooling the joining part and the transparent part together, to join the joining part and the transparent part together, in particular in a sterile sealed and / or hermetically sealed state, to obtain the internal component.

[0061] In other words, in step bb) and correspondingly in step dd), the joining part (or joining part and transparent part) can be subjected to positive or negative thermal expansion, which is carried out in such a way that a temperature-dependent thermal expansion difference is achieved between the joining part and the transparent part, with positive thermal expansion (expansion) being carried out in particular in step bb) and negative thermal expansion (contraction) being carried out in step dd).

[0062] On the other hand, it is also possible to precede in step a) the following method steps, in particular in this order, in order to prepare the internal component: aa) providing a transparent part and a joining part, in particular a joining part made of the same material as the transparent part; bb) obtaining the internal component by bringing the connecting part close to and / or pressing it against the transparent part at the front side, in order to connect the connecting part and the transparent part in a particularly sterile and / or hermetically sealed state, cc) preferably laser welding both parts together to obtain the internal component by tightly joining the joining part and the transparent part, in particular with a sterile seal and / or hermetic seal.

[0063] The overall method sequence for producing a biocompatible composite element further preferably includes, particularly as the last step, sterilization, particularly autoclaving, of the composite element comprising the outer frame and the internal components housed therein in a sterile sealed and / or hermetically sealed state.

[0064] The invention further relates to a bioreactor, in particular a disposable bioreactor, for culturing microorganisms or animal or plant cells, comprising a composite element as described above, e.g. polymer-to-polymer welded, attached as a connecting fitting to the wall of the bioreactor.

[0065] The bioreactor, in particular the single-use bioreactor, has composite elements attached to the wall as connecting fittings, in particular in a permanently fixed manner, for example by welding. The composite elements or the connecting fittings can in particular have the above-mentioned features.

[0066] In particular, the composite element comprises an outer casing made of or consisting of a polymer material, which outer casing is attached to the wall of the bioreactor and in particular is connected thereto in a sterile sealed and / or hermetically sealed manner, and further comprises an internal component made of or consisting of a transparent material, such as glass or glass ceramic, which is housed in a particularly permanently fixed manner in the outer casing in a sterile sealed and / or hermetically sealed manner and forms a window in the bioreactor, thereby allowing in particular spectral process control from outside the bioreactor.

[0067] The outer shell is preferably tubular with a first tube end and a second tube end and is attached to the wall of the bioreactor, e.g., by the first tube end, and preferably includes a flange that is attached to the wall of the bioreactor, the flange being preferably located between the first tube end and the second tube end or at the first tube end.

[0068] Bioreactors, in particular those designed for the containment of fluid media containing biological material, in particular those designed as disposable bioreactors, preferably comprise or consist of plastic, in particular sterilizable plastic.

[0069] The bioreactor is preferably autoclavable together with the composite elements attached to it.

[0070] A bioreactor with a composite element attached as a connection fitting to the wall of the bioreactor preferably has the features of any one of claims 1 to 11 with respect to the composite element.

[0071] Finally, the present invention also relates to a method for growing or culturing biological material, in particular microorganisms or cells, in a bioreactor, in particular a disposable bioreactor as described above, equipped with a biocompatible composite element, in particular a biocompatible composite element as described above and / or according to any one of claims 1 to 11, attached as a connecting fitting to the wall of the bioreactor, in which spectral process control is performed from outside the bioreactor through a window formed, in particular a physical, chemical or biological measurement parameter is obtained from outside the bioreactor through the window formed.

[0072] A preferred method for growing or culturing biological material includes introducing a fluid, particularly a biological material or a precursor of a biological material, into a bioreactor as described herein and capturing a measured physical, chemical or biological parameter using a window formed by a connection fitting as described above.

[0073] Preferably, the growth or culture methods disclosed herein include the production of pharmaceuticals, particularly biopharmaceuticals.

[0074] Preferably, the bioreactor is sterilized or autoclaved, especially after permanently attaching the connection fittings. Measurements can then be taken from outside the bioreactor. Preferably, spatially resolved measurements can be taken, e.g., to provide spatially resolved insight into metabolic processes.

[0075] The method of growing or culturing biological material may comprise measuring the radiation intensity and / or wavelength of electromagnetic radiation inside the bioreactor from outside the bioreactor, in particular in a spatially resolved manner.

[0076] If the bioreactor is irradiated from outside the bioreactor with electromagnetic radiation of a predetermined wavelength, preferably 250 nm, for a predetermined period of time, and after this irradiation the emission intensity and / or wavelength of the electromagnetic radiation inside the bioreactor is measured broadband or selectively at a wavelength, in particular at 270 nm, then fluorescently emitted portions of the light can be detected and analyzed with respect to predetermined metabolic processes within the photobioreactor.

[0077] Thus, for example, it is possible to envisage irradiating the bioreactor from outside the bioreactor with electromagnetic radiation of a given wavelength, preferably 250 nm, for a given period of time, and after this irradiation measuring from outside the radiation intensity and / or wavelength of the electromagnetic radiation inside the bioreactor in a broad band or selectively at a wavelength, in particular at 270 nm.

[0078] Phototrophic or mixotrophic microorganisms, particularly microalgae, yeasts and bacteria, that have been altered by mutagenesis may also be used in the methods disclosed herein.

[0079] In the following, some specific exemplary embodiments of the present invention, in particular glass-to-polymer sealing (GTPS), which should not be understood as being definitive, will be described with reference to the accompanying drawings, in which: FIG. [Brief explanation of the drawings]

[0080] [Figure 1] 1 is a cross-sectional view of a composite element according to a first embodiment. [Figure 2] FIG. 4 is a cross-sectional view of a composite element according to a second embodiment. [Figure 3] FIG. 10 is a cross-sectional view of a composite element according to a third embodiment. [Figure 4] FIG. 10 is a cross-sectional view of a composite element according to a fourth embodiment. [Figure 5] FIG. 10 is a cross-sectional view of a composite element according to a fifth embodiment. [Figure 6] FIG. 13 is a cross-sectional view of a composite element according to a seventh embodiment. [Figure 7] FIG. 13 is a cross-sectional view of a composite element according to a seventh embodiment. [Figure 8] 10A and 10B are cross-sectional and detailed views of a composite element according to a sixth embodiment. [Figure 9] 10A and 10B are cross-sectional and detailed views of a composite element according to a sixth embodiment. [Figure 10] 13A and 13B are cross-sectional and detailed views of a composite element according to an eighth embodiment. [Figure 11] 13A and 13B are cross-sectional and detailed views of a composite element according to an eighth embodiment. [Figure 12] 13A and 13B are cross-sectional and detailed views of a composite element according to a ninth embodiment. [Figure 13] 13A and 13B are cross-sectional and detailed views of a composite element according to a ninth embodiment. [Figure 14] 1A and 1B are perspective and top views of the surface of a bonded part having a microstructure. [Figure 15] 1A and 1B are perspective and top views of the surface of a bonded part having a microstructure. [Figure 16] FIG. 1 is a perspective view of a composite element mounted in a bioreactor according to a first embodiment. [Figure 17] FIG. 1 is a perspective view of a composite element mounted in a bioreactor according to a first embodiment. [Figure 18] FIG. 1 is a cross-sectional view of a composite element installed in a bioreactor according to a first embodiment. [Figure 19] FIG. 2 is a perspective detail view of a composite element according to the first embodiment. [Figure 20] FIG. 20 is a side view of a composite element according to a tenth embodiment. [Figure 21] 22A and 22B are cross-sectional views of a composite element according to a tenth embodiment (along plane AA in FIG. 22A and 22B). [Figure 22] 20A and 20B are cross-sectional views (in plane BB of FIG. 20) of a composite element according to a tenth embodiment. [Figure 23] FIG. 19 is a perspective view of a composite element according to a tenth embodiment. [Figure 24] FIG. 19 is a perspective view of a composite element according to a tenth embodiment.

[0081] The composite element (10) shown in Figures 1 and 19 comprises an outer frame (20) made of a polymer, such as polyetheretherketone (PEEK), and an inner component (30) formed as a one-piece transparent part (32) made of a transparent material, such as glass.

[0082] This embodiment is a polymer incorporation under pressure and can be produced, for example, as follows: the transparent part (32), in particular glass, is fitted into the polymer housing (20) under compressive stress.

[0083] For this purpose, the outer frame (20) formed as a polymer molded part is heated (for example to 200°C for PEEK), and the transparent part (32) formed as a plate in this example is inserted with a diameter larger than the inner diameter of the port and allowed to cool, thereby achieving an autoclavable, sterile, sealed assembly into the polymer under pressure.

[0084] The polymer shell 20 includes a flange 22 located at the first (proximal) tube end 20p, although the flange 22 can be located elsewhere, particularly between the first tube end 20p and the second tube end 20d, as shown in FIG.

[0085] 2, the composite element (10) comprises an outer frame (20) made of a polymer, e.g., polyethylene (PE), and a two-piece inner component (30) comprising a transparent part (32), e.g., a plate-like part made of glass, and a tubular connecting part (34), e.g., made of metal, e.g., 1.4404. In this embodiment, the connecting part (34) surrounds the transparent part (32).

[0086] This embodiment is a glass / metal / polymer composite and can be manufactured, for example, as follows: a transparent part (32) is pressure-sealed and / or hermetically sealed in a mating part (34), which in this example is formed as a profiled hollow metal cylinder. The mating part (34) or the internal component (30) thus formed is then sterile-sealed and covered with a polymer, such as polyethylene (PE), polypropylene (PP), polyamide (PA), polyethylene terephthalate (PET), ethylene vinyl alcohol (EVOH), polyether ether ketone (PEEK), polyaryl ether ketone (PAEK), polysulfone (PSU), polyphenylene sulfide (PPS), or a multi-polymer molded article containing multiple polymers, in the dimensions desired for the part, including the flange (22). Therefore, the outer frame (20) may comprise, consist of, be made from, or be manufacturable from one or more of the above materials.

[0087] The polymer shell (20) is provided with a flange (22) at the first tube end (20p), but the flange (22) may alternatively be provided at another location, particularly between the first tube end (20p) and the second tube end (20d), as shown in Figures 6 and 7.

[0088] 6 and 7 further comprises a multi-piece (here, three-piece) internal component (30) in which a connecting piece (34) is directly connected to the outer frame (20) and indirectly connected to the transparent piece (32). The connecting piece (34) is connected at its front to a frame (33) around the transparent piece (32), such that the transparent piece (32) is contained within the frame (33) in a sterile sealed, preferably hermetically sealed state, and the frame (33) is also connected to the connecting piece (34) in a sterile sealed, preferably hermetically sealed state, and the connecting piece (34) is also contained within the outer frame (20) in a sterile sealed, preferably hermetically sealed state.

[0089] The composite element 10 shown in Figure 3 also comprises a polymer outer frame 20 and a two-piece inner component 30, which comprises a transparent part 32 and a tubular connecting part 34. In this embodiment, the connecting part 34 is formed as a hollow cylinder made of a transparent material, in particular glass. Furthermore, the connecting part 34 in this case abuts the transparent part 32 on its front side.

[0090] This embodiment is a glass / glass / polymer composite and can be produced, for example, as follows: a joining part (34), in particular formed as a glass cylinder, is ground and polished on the front side, and a transparent part (32), for example formed as a plate, is ground and polished, and these parts are ground and polished to produce a coherent surface.

[0091] The two parts are then bonded together (i.e., the plate is pressed against the glass cylinder) so that they adhere to one another through intermolecular forces. The two parts are then firmly joined, for example by laser welding, to create a permanent, sterile-sealed bond. The internal component (30) thus formed is then covered with a polymer, again, for example, polyethylene (PE), in the dimensions desired for the part, including the flange (22), in a sterile-sealed manner.

[0092] This covering results in a polymeric shell 20, which in this embodiment includes a flange 22 at the first (proximal) tube end 20p, although the flange 22 could be formed elsewhere, particularly between the first tube end 20p and the second tube end 20d, as shown in Figure 8.

[0093] Figure 9 shows a further detailed view of the composite element (10) shown in Figure 8, in which the transparent part (32) protrudes from the outer frame (20) to form an overhang U. Since the contact angle of the material of the outer frame (20) with respect to the material of the transparent part is less than 90°, an angle α of less than 90° results between the outer frame (20) and the transparent part (32) accordingly.

[0094] The composite element (10) shown in Figure 4 comprises a polymer outer frame (20) and a multi-piece inner component (30) that includes a transparent piece (32), a frame (33) surrounding the transparent piece (32), and a tubular connecting piece (34). The connecting piece (34) is connected at its front to the frame (33) surrounding the transparent piece (32) such that the transparent piece (32) is received within the frame (33) in a sterile sealed, preferably hermetically sealed, condition, and the frame (33) is also connected to the connecting piece (34) in a sterile sealed, preferably hermetically sealed condition, and the connecting piece (34) is also received within the outer frame (20) in a sterile sealed, preferably hermetically sealed condition.

[0095] In this embodiment, the internal component 30, in particular the transparent part 32 and the frame 33, protrudes at least partially from the outer frame 20. However, it is also conceivable that the outer frame 20 could alternatively completely surround the internal component 30 over its entire length.

[0096] The outer frame 20 also has a flange 22 at the first tube end 20p in this embodiment, but the flange 22 can also be provided at another location, in particular between the first tube end 20p and the second tube end 20d, as shown in Figures 10 and 12.

[0097] Figures 11 and 13 show further details of the composite element (10) shown in Figures 10 or 12. An angle α of less than 90° is assumed between the outer frame (20) and the inner component (30). For this purpose, an arch-shaped recess (23) is introduced in the outer frame (20). This recess (23) is present in the polymer material at the material transition to the inner component (30).

[0098] Figures 14 and 15 show the surface of a joining part into which microstructures in the form of numerous protrusions (36) have been introduced. A first group of grooves (37') intersects with a second group of grooves (37''), resulting in numerous protrusions (36) between the grooves.

[0099] 16, 17, and 18 show a bioreactor wall (40) as part of a bioreactor. The bioreactor wall (40) has penetrations (42) (also referred to as ports (42)) present therein, which form openings to the interior of the bioreactor, and a composite element (10) extending at least partially through the opening formed by the penetrations (42). In this example, the composite element (10) is formed in accordance with the composite element (10) shown in FIG. 1. However, it should be understood that any of the above composite elements (10) could extend through the penetrations (42) shown here.

[0100] A measuring probe 50 is installed inside the composite element 10, allowing measurements to be taken inside the bioreactor through the transparent part 32 of the composite element 10. A sealing element 44, e.g., an O-ring, holds the composite element 10 in a fluid-tight manner, particularly preferably in a sterile and / or hermetically sealed manner, particularly by positive and frictional contact with the penetration 42 in the wall 40 of the bioreactor. The measuring probe 50 is also frictionally connected to the composite element 10 by a friction element 46, preferably an O-ring, which is held in a cylindrical recess of the composite element by a substantially annular pressure element 48, which applies a predetermined, adjustable force to the friction element 46 in the axial direction of the composite element 10.

[0101] The composite element (10) is held in a fixed, but removable, position in the penetration (42) in the wall (40) of the bioreactor by a union nut (52), which is in particular formed as a cap nut, but which is held in a non-removable manner on the composite element (10) by a retaining ring or snap ring (54) so ​​that it can rotate but with only a small amount of axial play.

[0102] Figures 20-24 show various views of a biocompatible composite element (10) comprising an outer frame (20) made of or including a polymeric material, such as PE, and a multi-piece inner component (30).

[0103] The outer frame 20 is tubular and has a flange 22 between the two tube ends 20p, 20d. In this embodiment, the flange is closer to the second tube end 20d than to the first tube end 20p, although this is generally applicable to any embodiment. At the first tube end 20p, the outer frame 20 has a thread 24, which is also generally applicable to any embodiment.

[0104] The internal component (30) is formed in three pieces in this embodiment, and includes a tubular connecting piece (34) made of or containing a metal, for example, austenitic-ferritic duplex stainless steel, such as 1.4462. At the second tube end (20d), a transparent piece (32), for example, made of or consisting of sapphire, is housed within the connecting piece (34). In this case, the transparent piece (32) is directly connected to a surrounding frame (33), which is made of or from, for example, glass and is directly connected to the connecting piece (34). The transparent element 32 is thus accommodated in the frame 33 in a sterile, preferably hermetically sealed state, and the frame 33 is also joined to the connecting element 34 in a sterile, preferably hermetically sealed state, which is also accommodated in the outer frame 20 in a sterile, preferably hermetically sealed state, whereby the transparent element 32 is joined to the connecting element 34 in a sterile, preferably hermetically sealed state, and the connecting element 34 is also accommodated in the outer frame 20 in a sterile, preferably hermetically sealed state. As mentioned above, the frame 33 may comprise or consist of, for example, glass, whereby a pressed product having or consisting of glass powder is conceivable. In this case, the biocompatible composite element 10 can be or has been produced, for example, by heating in a conveyor oven, resulting in a glass-metal composite.

Claims

1. A biocompatible composite element (10) for use as a connection fitting for a bioreactor, for forming a window in said bioreactor, said biocompatible composite element (10) comprising: an outer frame (20) for attachment to the wall of the bioreactor, the outer frame (20) comprising or consisting of a polymer material; an internal component (30) having or consisting of a transparent material; It is equipped with the internal components are inseparably housed within the outer shell in a sterile, sealed condition and form a window that allows for spectral process control from outside the bioreactor; The internal component (30) is formed of a plate-shaped transparent part (32) and a tubular connecting part (34), The joining part (34) has a profile (35) for increasing the contact area with the outer frame (20) and has a microstructure with a number of protrusions (36); The internal component (30) is formed of at least two pieces, namely the transparent part (32) and the connecting part (34), The connecting part (34) connects the transparent part (32) and the outer frame (20) together directly or indirectly. A biocompatible composite element (10).

2. the outer frame (20) is annularly shaped so that the outer frame surrounds the internal component in cross section; and / or the outer frame (20) is tubular with first and second tube ends (20p, 20d), the formation being such that the first tube end (20p) allows the outer frame (20) to be attached to the wall of the bioreactor; or the outer shell comprises a flange (22) for attachment to the wall of the bioreactor, the flange (22) being located between the first and second tube ends or at the first tube end (20p); and / or The outer frame includes a threaded portion (24), the threaded portion (24) being disposed on the first pipe end (20p). The biocompatible composite element (10) of claim 1.

3. The connecting part (34) is directly or indirectly connected to the transparent part (32) and the outer frame (20), The transparent element (32) is disposed at the second tube end (20d), and the connecting element (34) extends to the first tube end (20p). A biocompatible composite element (10) according to claim 1 or 2.

4. the connecting part (34) surrounds the transparent part (32) in cross section, such that the transparent part (32) is contained within and connected to the connecting part (34) in a sterile sealed state, and the connecting part (34) is also inseparably contained within the outer frame (20) in a sterile sealed state; and / or the joining part (34) comprises or consists of metal, and / or The connecting part (34) surrounds in cross section a frame (33) around the transparent part (32), the frame (33) having or consisting of glass, such that the transparent part (32) is contained in the frame (33) in a sterile sealed state, the frame (33) is also contained in the connecting part (34) in a sterile sealed state, and the connecting part (34) is also contained in the outer frame (20) in a sterile sealed state. A biocompatible composite element (10) according to claim 3.

5. the connecting element (34) is in contact with the transparent element (32) at its front surface and / or is connected to the transparent element (32) at its front surface, said contact or connection being effected such that both the transparent element (32) and the connecting element (34) are housed within the outer frame (20) in a sterile sealed state; and / or the coupling element (34) comprises or consists of a transparent material, including glass, sapphire, ceramic or glass-ceramic; or The joining element (34) comprises or is made of an opaque material, including ceramic or metal, an oxidizable metal, or aluminum, and the joining element (34) has an oxide layer on one surface adjacent to the transparent element (32). A biocompatible composite element (10) according to claim 3.

6. The connecting part (34) is connected at the front side to the transparent part (32) and / or the frame (33) around the transparent part (32), and the connection is the transparent element (32) is contained in the frame (33) in a sterile sealed state, the frame (33) is also connected to the connecting element (34) in a sterile sealed state, and the connecting element (34) is also contained in the outer frame (20) in a sterile sealed state; and / or The transparent part (32) is connected to the connecting part (34) in a sterile sealed state, and the connecting part (34) is also contained in the outer frame (20) in a sterile sealed state. carried out and / or The internal component (30) at least partially protrudes from the outer frame (20). A biocompatible composite element (10) according to claim 3.

7. the internal components (30) are inseparably housed within the outer frame (20) under compressive stress; and / or The composite element (10) is manufactured or can be manufactured by expanding the outer frame (20) relative to the inner component (30), then inserting the inner component (30) into the outer frame (20), and then shrinking the outer frame (20) relative to the inner component (30). A biocompatible composite element (10) according to any one of claims 1 to 6.

8. the internal components (30) are housed in the outer frame (20) in a stress-neutral state; and / or the composite element (10) is manufactured or can be manufactured by attaching the outer frame (20) to the inner component (30) from the outside by supplying a liquid polymer material to the inner component (30) from the outside and then curing it, The material of the outer frame (20) and the material of the inner component (30) form a contact angle of less than 90°. A biocompatible composite element (10) according to any one of claims 1 to 6.

9. the transparent part (32) is contained within the connecting part (34) under compressive stress, and / or The internal component (30) is manufactured or can be manufactured by expanding the connecting part (34) relative to the transparent part (32), then inserting the transparent part (32) into the connecting part (34), and then shrinking the connecting part (34) relative to the transparent part (32). A biocompatible composite element (10) according to claim 7 or 8.

10. the transparent part (32) is bonded to the bonding part (34) in a stress-neutral manner, and / or The internal component (30) is manufactured or can be manufactured by bringing the connecting part (34) close to and / or pressing it against the transparent part (32) at the front side, and then laser welding the connecting part (34) and the transparent part (32). A biocompatible composite element (10) according to claim 7 or 8.

11. A method for manufacturing a biocompatible composite element (10) according to claim 7, comprising the steps of: Providing an outer frame (20) having or consisting of a polymer material and an inner component (30) having or consisting of a transparent part (32) including glass, sapphire, or glass ceramic; causing a relative expansion of the outer frame (20) with respect to the inner component (30), the expansion being achieved by heating the outer frame or by heating the outer frame and the inner component together; inserting the internal component (30) into the external frame (20) which has been expanded against the internal component (30); causing relative shrinkage of the outer frame (20) relative to the inner component (30), the shrinkage being achieved by cooling the outer frame or by cooling the outer frame and the inner component together, so that the outer frame (20) encloses the inner component (30) in a sterile sealed state; A method comprising:

12. A method for manufacturing a biocompatible composite element (10) according to claim 8, comprising the steps of: providing an internal component (30) and a liquid polymer material for forming an outer frame, wherein the polymer material and the material of the internal component (30) form a contact angle of less than 90°; supplying the liquid polymer material from the outside to the internal component (30), to the joining part (34) and / or the transparent part (32); hardening the liquid polymer material to form an outer shell (20) enclosing the internal components (30) in a sterile seal; A method comprising:

13. 13. A method according to claim 12 for producing a biocompatible composite element (10) according to claim 9, comprising: Providing a transparent part (32) and a joining part (34); causing a relative expansion of the joining element (34) with respect to the transparent element (32), the expansion being achieved by heating the joining element or by heating the joining element and the transparent element together; Inserting the transparent part (32) into the joining part (34) which is expanded relative to the transparent part (32); causing relative shrinkage of the joining part (34) relative to the transparent part (32), and effecting said shrinkage by cooling the joining part (34) or by cooling the joining part and the transparent part together, thereby bonding the joining part (34) and the transparent part (32) to obtain the internal component (30); performing the method steps of claim 12; A method comprising:

14. Providing a transparent part (32) and a joining part (34); a step of bringing the joining part (34) close to or pressing the transparent part (32) at the front surface thereof to join the joining part (34) and the transparent part (32) together to obtain the internal component (30); performing the method steps of claim 12; 13. A method according to claim 12 for producing a biocompatible composite element (10) according to claim 10, comprising:

15. 15. A method according to any one of claims 11 to 14 for producing a biocompatible composite element (10), comprising: Sterilizing the composite element (10) comprising the outer frame (20) and the inner component (30) contained therein in a sterile sealed state. A method comprising:

16. A bioreactor for cultivating microorganisms or cells, comprising a composite element (10) according to any one of claims 1 to 10 attached as a connection fitting to the wall of the bioreactor, The composite element comprises an outer frame (20) having or consisting of a polymer material, the outer frame (20) being attached to the wall of the bioreactor and being connected thereto in a sterile, sealed manner; and The composite element comprises an internal component (30) having or consisting of a transparent material, including glass, sapphire, or glass ceramic, the internal component being housed within the outer shell (20) in a sterile sealed condition and forming a window within the bioreactor, thereby allowing spectral process control from outside the bioreactor; The outer frame (20) is tubular with first and second tube ends (20p, 20d) and is attached to the wall of the bioreactor by the first tube end (20p), or the outer frame has a flange (22) and is attached to the wall of the bioreactor by the flange (22), the flange (22) being located between the first and second tube ends (20p, 20d) or at the first tube end (20p). Bioreactor.

17. 17. A method for growing or culturing biological material in a bioreactor according to claim 16, comprising a biocompatible composite element (10) according to any one of claims 1 to 10 attached as a connecting fitting to the wall of the bioreactor, including the production of pharmaceuticals, including biopharmaceuticals, wherein spectral process control is performed from outside the bioreactor and physical, chemical or biological measurement parameters are known from outside the bioreactor through formed windows.

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