Cassette system for aseptic mixing processes

Abstract

The present invention provides a frame (10) for holding and positioning a static mixing device (16) along with flexible containers (11, 12, 13, 14) for liquid substrates, waste products, and liquid products. The frame includes a sealable area for holding the containers and flexible containers, and a mixing device. Other aspects relate to kits including the frame along with one or more matching components, such as a static mixing device and flexible containers, as well as apparatus for operating the frame in its assembled state, and methods involving use of the frame.

Description

[Background technology]

[0001] explanation Process automation and upscaling and downscaling of mixing processes are tasks that are routinely addressed in various fields of biotechnology and medicine.

[0002] Systems for the production of mixed fluids or products based on the combination of two or more components require a source of liquid raw materials, feed lines, a chamber where the mixing or reaction takes place, and an outlet for harvesting the product. The chamber often represents the heart of such a system and can be bulky, such as a vessel, or miniaturized, as in microfluidic approaches. However, the configuration of the system, conditions, and raw material quality also play a crucial role in these processes.

[0003] An example of a discontinuous mixing system is the device for mixing, storing, and homogenizing liquids, as disclosed in U.S. Patent No. 7,784,997. It comprises a rigid container fitted with a non-invasive pump. The container encloses a disposable bag with an orifice on the bottom surface used as a liquid outlet and additional orifices at the top of the bag for adding various liquids to produce the mixture. One of the top orifices is used (with the aid of a pump) to return the liquid to the inside of the bag, allowing for closed-loop circulation. This system is intended for disposable use, avoiding the cleaning and sterilization steps required when rigid tanks are used for mixing. It can handle bags with volumes of 25 to 3,000 L, but does not appear to be suitable for producing smaller volumes, such as μl or ml volumes.

[0004] EP 1146959 discloses an apparatus for the continuous production of encapsulated therapeutic compounds, providing an example of a precisely controlled metering system. The apparatus includes a pressurized transfer means for transferring the phases to a lipid phase storage means and an aqueous phase storage means, preferably a mixing device, such as a static mixer. The apparatus of EP 1146959 further includes a premixing system. The phases are transported from the storage means to the premixing and mixing chamber with the aid of a metering pump driven by a motor. Because this system is continuous, it can produce larger quantities of the desired product.

[0005] Despite these and other solutions to mixing systems and methods, there remains an incomplete need for methods, devices, and apparatus for producing small quantities of products that are fast, reliable, and can be performed under standardized conditions, particularly for applications in the field of personalized medicine.

[0006] It is therefore an object of the present invention to provide means, devices, components, and methods for the production of mixed fluids or products based on the combination of two or more components that are flexible in terms of adapting to different types of products, yet robustly, quickly, and reliably produce the desired product. They also aim to provide a high-throughput approach. Further objects of the present invention will become apparent based on the following description of the invention, the drawings, and the claims. [Prior art documents] [Patent documents]

[0007] [Patent Document 1] U.S. Patent No. 7,784,997 [Patent Document 2] European Patent No. 1146959 Summary of the Invention

[0008] In a first aspect, the present invention relates to a frame adapted to simultaneously hold (a) first and second flexible substrate containers, each having an outlet port, (b) a flexible waste container having an inlet port, (c) a flexible product container having an inlet port, (d) a static mixing device having at least first and second inlet ports and an outlet port, and (e) conduits for fluidly connecting the outlet port of the first substrate container with the first inlet port of the static mixing device, the outlet port of the second substrate container with the second inlet port of the static mixing device, and the outlet port of the static mixing device with the inlet ports of the waste container and the product container. The frame is further characterized by comprising a first sealable area for holding the first flexible substrate container, a second sealable area for holding the second flexible substrate container, and means for holding the static mixing device. Further, the frame has an operational orientation, and the means for holding the first and second sealable regions and the static mixing device are arranged such that fluid flow from the first and second substrate containers to the static mixing device and / or from the static mixing device to the waste or product container in the operational orientation occurs at least partially in an anti-gravity direction. The frame is particularly useful for aseptic small-batch production of sterile liquid products, particularly those requiring mixing of two fluid substrates without the use of pumps.

[0009] In a further aspect, the present invention provides a kit comprising a frame as described herein and any one of the following components: (a) a first and / or second flexible substrate container, (b) a flexible waste container, (c) a flexible product container, (d) a static mixing device, and / or (e) one or more conduits for fluidly connecting the outlet port of the first substrate container with the first inlet port of the static mixing device, the outlet port of the second substrate container with the second inlet port of the static mixing device, and the outlet port of the static mixing device with the inlet ports of the flexible waste container and the flexible product container. The kit may also comprise the frame and all of components (a) through (e), including both the first and second flexible substrate containers and all conduits for the specified fluid connections. Alternatively, the kit may comprise components (a) through (e) but not the frame.

[0010] According to a further aspect of the present invention, there is provided a flexible container adapted for use as a flexible substrate container, flexible waste container, or flexible product container. The container has an interior space for holding a fluid material surrounded by a flexible front wall and a flexible rear wall, each wall being made of a polymeric material. The container further comprises at least one inlet or outlet port for allowing fluid communication with the interior space. The flexible front wall and the flexible rear wall are connected to each other to form a sealed edge that substantially encloses the interior space. The edge includes four corner regions so that the interior space has an overall square or rectangular shape when empty. The container is further characterized by having at least two through-holes provided in the sealed edge, a first through-hole at or near a first corner region of the edge and a second through-hole at or near a second corner region of the edge, the second corner region adjacent to the first corner region. Still further aspects of the present invention relate to an apparatus for aseptically filling first and second flexible substrate containers assembled in a frame, an apparatus for mixing first and second liquid substrates, wherein the liquid substrates are contained in first and second flexible substrate containers, and the containers are assembled in a frame, and a method for mixing first and second liquid substrates, which method relies on the use of a frame in which the first and second flexible substrate containers are assembled.

[0011] Further aspects, preferred and optional features and embodiments are described in more detail below. [Brief explanation of the drawings]

[0012] [Figure 1] FIG. 2 is a perspective view of the frame (10). [Figure 2] FIG. 2 is a front view of the frame (10) of FIG. [Figure 3] FIG. 3 is a front view of the frame (10) of FIG. 2 without assembled parts. [Figure 4] FIG. 2 is a perspective view of the frame (10) of FIG. 1 without assembled parts. [Figure 5] FIG. 2 is a perspective view of the frame (50). [Figure 6] FIG. 6 is a top view of the frame (50) shown in FIG. [Figure 7] FIG. 7 is a top view of the frame (50) shown in FIG. 6 without the assembled parts. [Figure 8] FIG. 2 is a perspective view of the frame (80). [Figure 9] FIG. 9 is a top view of the frame (80) of FIG. 8. [Figure 10] FIG. 9 is a perspective view of the frame (80) of FIG. 8 without assembled parts. [Figure 11] FIG. 10 is a top view of the frame (80) of FIG. 9 without assembled parts. [Figure 12] FIG. 1 is a front view of the frame (120). [Figure 13] FIG. 1 is a diagram showing an example of a flexible container (1). [Figure 14] FIG. 14 is a perspective side view of the flexible container (1) of FIG. [Figure 15] FIG. 10 is a front view, or user-facing side view, of another example of a frame (150) with the parts assembled. [Figure 16] FIG. 16 is a front view of the rear surface of the frame (150) shown in FIG. [Figure 17] FIG. 16 is a front elevational view of the frame (150) shown in FIG. 15 without the assembled parts. [Figure 18] FIG. 18 is a front view of the rear surface of the frame (150) shown in FIG. [Figure 19] FIG. 16 is a front perspective view of the frame (150) shown in FIG. [Figure 20] FIG. 20 is a perspective view of the rear face of the frame (150) shown in FIG. 19. [Figure 21] 17 is a detailed or enlarged view of a portion of the rear face of the frame (150) shown in FIG. 16. [Figure 22] FIG. 17 is a perspective view of an exemplary static mixing device (162) that may be used in combination with the frame (150) or kit shown in, for example, FIGS. 15 and 16. [Figure 23] FIG. 23 is a perspective view of the static mixing device (162) shown in FIG. 22. [Figure 24] FIG. 10 is a front perspective view of another example of a frame (240). [Figure 25] FIG. 25 is a perspective view of the rear face of the frame (240) shown in FIG. 24. DETAILED DESCRIPTION OF THE INVENTION

[0013] In one aspect, the present invention provides a frame adapted to simultaneously hold (a) first and second flexible substrate containers, each having an outlet port, (b) a flexible waste container having an inlet port, (c) a flexible product container having an inlet port, (d) a static mixing device having at least first and second inlet ports and an outlet port, and (e) conduits for fluidly connecting the outlet port of the first substrate container with the first inlet port of the static mixing device, the outlet port of the second substrate container with the second inlet port of the static mixing device, and the outlet port of the static mixing device with the inlet ports of the waste container and the product container. The frame is further characterized by comprising a first sealable area for holding the first flexible substrate container, a second sealable area for holding the second flexible substrate container, and means for holding the static mixing device. Further, the frame has an operational orientation, and the means for holding the first and second sealable regions and the static mixing device are arranged such that fluid flow from the first and second flexible substrate containers to the static mixing device and / or fluid flow from the static mixing device to the waste or product container in the operational orientation occurs at least partially in an anti-gravity direction.

[0014] The inventors have found that such a frame is particularly useful for aseptically mixing or reacting two fluid substrates in small batches to obtain a sterile liquid product, such as a liquid pharmaceutical. For example, the mixing or reaction of the two substrates may be driven by pressure, such as pressurized gas, enabling a process in which all substrate or product-contacting components can be provided as sterile disposables. Furthermore, the frame may be used to minimize dead volume, which can result in low product yields, especially in very small batches such as in personalized pharmaceutical manufacturing. The at least partial anti-gravity flow of fluids provided by the frame reduces the adverse effects of gases that may be present in the substrate container or that may be generated during the mixing process, such as bubble formation. These and additional unexpected effects and advantages are described in more detail below.

[0015] As used herein, a frame should be understood as a support structure or holder for particular components and provides a spatial arrangement thereof. In this context, those skilled in the art will understand that the expression "for," such as "for holding," implies that the frame is arranged or adapted for a particular purpose.

[0016] Flexible substrate containers, also referred to herein simply as substrate containers, are containers adapted to hold fluid substrates, particularly liquid substrates, such as sterile liquids that are intermediates in the preparation of injectable pharmaceutical products. They may exhibit a high degree of flexibility, similar to infusion bags. Each substrate container comprises at least one outlet port that can be fluidly connected to a static mixing device via a conduit.

[0017] Similarly, the waste container and the product container are adapted to hold fluid waste or fluid product, respectively, and are also flexible. Each of these containers has at least one inlet port fluidly connectable to the static mixing device, for example, via a conduit, so that the container can receive the waste or product from the mixing device. Also, at least a portion of the product and waste are typically liquid. In an alternative embodiment, the waste container and / or the product container are not flexible, but the substrate container is flexible.

[0018] With respect to conduits, it should be understood that they may have any structure that allows for the conduction of fluid materials, such as tubular or pipe-like structures. At the same time, conduits should not be understood as requiring a specific minimum length. Indeed, according to some embodiments, the length of a conduit may be very small even relative to its outer diameter, thereby minimizing the internal volume and therefore potentially the dead space associated with the conduit.

[0019] The frame may be adapted to hold any type of static mixing device. As used herein, a static mixing device may be any mixing device with no moving parts.

[0020] As described above, the frame includes first and second sealable regions. As used herein, a sealable region of a frame is a structure adapted to hold or contain or contribute to the containment of an object, such as a flexible substrate container, and is sealable against at least one counterpiece to enclose such object within a sealed space.

[0021] For example, the sealable area may be shaped as a cavity. In this context, a cavity in the frame or in the tray portion of the frame refers to a portion of a surface that can be recessed or indented to receive one or more objects, such as a substrate container. Alternatively, the sealable area or cavity may be formed by a rim, such as a circumferential rim, disposed on or extending from the frame, rather than a recessed flat portion of the frame. To avoid doubt, the term "cavity" does not require the flexible container to fit completely into the respective cavity; particularly when filled with a fluid substrate, the flexible container may have a height that somewhat exceeds the height of the cavity that holds the container. In some preferred embodiments, the first and / or second sealable areas are defined by a circumferential rim extending from the frame.

[0022] Optionally, the circumferential rim is configured to sealingly receive a counter-piece having a cavity for accommodating a respective flexible substrate container. In some related embodiments, both the first and second sealable regions represent substantially flat regions of the frame extending from the frame and having a circumferential rim adapted to sealingly receive a counter-piece having a cavity for at least partially accommodating a respective flexible container. As will be understood by those skilled in the art, the extent to which the structure or shape of the sealable regions and the structure or shape of the counter-piece contribute to the overall shape of the sealed space in which the flexible substrate containers are enclosed may vary.

[0023] In a related embodiment, the frame may be designed to include a tray portion. A tray portion, in this context, may be broadly understood as a relatively flat structure, optionally interrupted by an opening, designed to hold one or more objects. Such a tray or tray portion may include first and second sealable regions in the form of cavities. As mentioned above, the cavities may be formed, for example, by recesses provided in the otherwise flat portion of the tray portion of the frame and / or by providing rims disposed on or extending from the tray. The frame, and optionally the tray portion of the frame, may further comprise means for holding a static mixing device. This means may also be formed as or include a cavity or recess, although other structures, such as rims or clips, may also be used.

[0024] According to this aspect of the invention, the frame has an operational orientation. In other words, it is configured to hold specific components in a specific orientation in which a process of mixing or reacting two substrates to obtain a product is carried out. As described above, the frame is configured such that, in its operational orientation, the first and second sealable regions or cavities and the means for holding the static mixing device are spatially arranged so that fluid flow from the first and second substrate containers to the static mixing device occurs at least partially in a direction opposite to gravity, or so that fluid flow from the static mixing device to the waste or product container occurs at least partially in a direction opposite to gravity, or both. In this context, the phrase "at least partially in a direction opposite to gravity" should be understood to mean that, for at least a portion of each flow path, the downstream end is higher than the upstream end, without requiring a flow direction exactly opposite to the direction of gravity (i.e., at an angle of 180° relative to the direction of gravity).

[0025] As will be apparent to those skilled in the art, and for the avoidance of doubt, the frame disclosed herein is not configured for use in the context of microfluidics or as part of a microfluidic system or device. Microfluidics refers to systems that manipulate very small volumes of fluid using small channels, ranging in size from tens to hundreds of micrometers. Furthermore, microfluidics utilizes fluid behavior at the microscale, which differs from "normal" or macrofluidic behavior in that factors such as surface tension, energy dissipation, and fluidic resistance dominate in the system. In contrast, the conduits that the frame is configured to hold, including the conduit fluidically connecting the outlet port of a first substrate container with the first inlet port of a static mixing device, the conduit fluidically connecting the outlet port of a second substrate container with the second inlet port of a static mixing device, and the conduits fluidically connecting the outlet port of a static mixing device with the inlet ports of a waste container and a product container, are non-microfluidic conduits. For example, their inner diameters are typically at least 1 millimeter, or several millimeters, e.g., in the range of about 1 mm to about 10 mm. Additionally, in some preferred embodiments, the conduits are flexible (e.g., flexible tubing) and reversibly connectable with the ports to which they are adapted for fluid connection. These characteristics further distinguish the present disclosure from microfluidic systems.

[0026] Thus, the static mixer is not a microfluidic mixer and is configured to mix fluids on a macrofluidic scale, even though this may be considered small in scale compared to typical large-scale pharmaceutical manufacturing processes. In some preferred embodiments, the static mixer is adapted to mix fluids at a total flow rate of about 10 mL to about 1,000 mL / min. In other embodiments, the static mixer is adapted to mix fluids at a total flow rate of about 20 mL to about 600 mL / min, or about 30 mL to about 300 mL / min, respectively.

[0027] Also, with respect to dimensions, and according to some further embodiments, the first and second sealable areas are shaped and sized to hold flexible substrate containers having internal volumes ranging from about 10 mL to about 3,000 mL. Also preferred are sealable areas shaped and sized to hold flexible substrate containers having internal volumes ranging from about 50 mL to about 1,500 mL, respectively. Other preferred substrate containers have internal volumes of about 300±100 mL, 500±200 mL, 1,000±300 mL, and 1,500±300 mL, respectively. In some related embodiments, the flexible product containers each have internal volumes ranging from about 50 mL to about 4,000 mL, or from about 100 mL to about 2,000 mL, or from about 300 mL to about 2,000 mL.

[0028] In some preferred embodiments, the first and second flexible substrate containers are different in size. For example, the internal volume of the first substrate container may be larger than the internal volume of the second substrate container. In some embodiments, the internal volume of the first substrate container is about 1.5 to about 4 times larger than the internal volume of the second substrate container. Thus, the shapes and dimensions of the first and second sealable areas are adapted to hold such substrate containers having different sizes. In some embodiments, the first and second sealable areas have substantially similar or identical heights (or lengths, depending on the operating orientation of the frame, as discussed below) and substantially different widths. For example, the width of the first sealable area may be, for example, 1.5 times or more larger than the width of the second sealable area.

[0029] In some further embodiments, the first and second sealable areas are positioned close to each other to minimize the dimensions of the frame and the length of the conduit required for liquid flow from the substrate container to the static mixer, thus reducing dead volume in the flow path. Optionally, the first and second sealable areas are located adjacent to each other, and the minimum distance between the first and second sealable areas is less than 10% of the width of the first sealable area.

[0030] In some embodiments, the frame further comprises means for holding or securing flexible product containers and / or flexible waste containers. For example, the means for holding these containers may be represented by cavities, and such cavities may be provided in the tray portion of the frame. Alternatively or additionally, hooks, protrusions, pins, or other means for holding or securing these containers may be provided. In some preferred embodiments, the means are adapted to allow reversible fixation of the respective containers to the frame. In this context, reversible fixation should be understood as fixation that is easily reversible for the user non-destructively, preferably without tools.

[0031] Furthermore, various options exist for positioning the means for holding or securing the flexible product container and / or flexible waste container. In some embodiments, the frame is configured to hold the product container and the waste container on the same side of the frame. In other embodiments, these two containers are attached to different sides of the frame. In some preferred embodiments, the frame has a front face facing the user in its operational orientation and a rear face opposite the front face, and the means for securing the flexible product container is located on the front face of the frame, and the means for securing the flexible waste container is located on the front or rear face of the frame. The advantage of this configuration is that the space for holding these containers provided by a frame having given dimensions is used more efficiently.

[0032] In this context, the front or user-facing side should be understood broadly regardless of the operational orientation of the frame. For example, for a frame with a vertical operational orientation, the front side is also oriented vertically, while for a frame with a horizontal operational orientation, the front side is also oriented horizontally and typically faces the top side of the frame.

[0033] In some preferred embodiments, the frame is adapted for a vertical operating orientation. This means that the cavities provided for holding the substrate containers and / or product and waste containers may not be sufficient to hold the respective containers in place during operation. Accordingly, the first and second sealable regions may each comprise one or more means for securing the respective flexible substrate containers.

[0034] In some preferred embodiments, one or more means for securing the flexible substrate containers are disposed on the front surface of the frame. Again, such means may be adapted to reversibly secure the respective containers, as described above in the relevant context. The means may optionally be shaped as hooks, protrusions, or pins arranged to receive the flexible substrate containers having through-holes positioned to match the hooks, pins, or protrusions. The pins may be, for example, barbed pins or snap-lock pins.

[0035] The options and preferences disclosed herein regarding the operational orientation of the frame and fixtures for the flexible substrate containers should be construed not only as separate individual disclosures, but also as combinations thereof. For example, it is clear that the present disclosure also provides a frame adapted for a vertical operational orientation having first and second sealable areas for holding first and second flexible substrate containers provided on a front surface of the frame, each sealable area comprising means for securing a respective substrate container, the means being arranged on the front surface of the frame to allow the substrate containers to be fixed to the frame at their front surfaces.

[0036] In some embodiments, the frame or tray portion of the frame includes not only cavities for the first and second substrate containers, but also a cavity for a flexible product container or a cavity for a flexible waste container, or both. Such embodiments require a significantly larger tray portion or tray area relative to the overall frame size. Due to these additional cavities, the tray portion may represent a larger frame portion than the rest of the frame. In some further embodiments, the entire frame may be shaped as a tray. Alternatively, the tray portion includes only two cavities for the containers, i.e., for the two designated substrate containers. In one embodiment, the tray portion of the frame includes only cavities for the first and second substrate containers, optionally including a cavity for a flexible product container.

[0037] In some embodiments, the first and second cavities for holding the first and second substrate containers are both located within a larger recess in the tray portion. The recess in this context may have the general shape of the tray cavity, so that the two cavities for individually holding the two substrate containers may be considered sub-cavities of the larger cavity. Furthermore, it is required that each of the two substrate containers be provided with an individual sealable area or cavity that is essentially sealable relative to the other cavity.

[0038] As described above, the first and second sealable regions may be shaped as cavities. While the cavities may, in principle, be shaped to accommodate and hold the first and second substrate containers, additional fixation of the containers may be useful for better handling. In some embodiments, the first and second cavities, or preferably each of the first and second cavities, comprise one or more means for fixing the respective flexible substrate containers. This provides the added advantage that the frame can be tilted while holding the containers without the containers falling out of the cavities. In some embodiments, the one or more means are adapted to reversibly fix the substrate containers, allowing for easy and non-destructive removal of the containers after use.

[0039] According to some further embodiments, at least two means for securing flexible substrate containers are provided in one or more of the sealable areas or tray cavities, such as two, three, or four means. One advantage of these embodiments is that each container can be more stably held in its desired position. A further advantage is that a user or operator can select a unique, specific spatial arrangement for each sealable area or cavity, which helps match each container with the correct sealable area or cavity.

[0040] For example, a fixing system may be used that includes protrusions or pins arranged in the peripheral zone of the flexible container, i.e., in the sealable area or cavity and through-holes provided at a distance from the lumen of the respective container. Advantageously, the positions of the protrusions or pins may coincide with the positions of the through-holes of the container, although the spatial arrangement may be different for each sealable area or cavity and corresponding container. As mentioned above, in some embodiments, the means for fixing the flexible substrate container are shaped as protrusions or pins for receiving a flexible substrate container having through-holes positioned to coincide with the pins or protrusions. In this particular situation, the protrusions or pins may be snap-fit ​​pins, such as pins with an arrowhead or mushroom-shaped head.

[0041] The spatial arrangement of the securing means may be used not only to ensure that the correct container is secured to the sealable area or inserted into the cavity, but also to facilitate its settling or insertion in the desired or correct orientation. Thus, in some embodiments, the means for securing each flexible substrate container is adapted to secure each flexible substrate container only when said container has the desired orientation.

[0042] In some embodiments, the flexible product container and / or flexible waste container, or only the flexible waste container, is reversibly secured directly to the frame itself by the securing means. In other words, the frame does not include a cavity or sealable area for holding the one or more containers. A similar or equivalent securing system to the systems used to hold the substrate containers described herein may be used, for example, a system comprising protrusions or pins disposed on non-recessed portions of the frame or frame portions, with through-holes provided in the peripheral zone of the flexible container. In some embodiments, the fasteners on the frame may be arranged so that the flexible waste container can be reversibly secured to a surface of the frame opposite the fasteners provided and arranged to secure the flexible substrate container and product container. As mentioned above, the means for securing the flexible product container may optionally be arranged on the front side of the frame, and the means for securing the flexible waste container may be arranged on the rear side of the frame.

[0043] Additional techniques may be used to ensure proper operation of the frame and the components it holds. In some embodiments, for example, the frame includes an identification tag. In some further preferred embodiments, the identification tag is a radio frequency identification (RFID) tag. Also preferred are embodiments in which one or both of the substrate containers have an identification tag, such as an RFID tag.

[0044] As described above, the frame may be configured to operate in a particular operating orientation. In some preferred embodiments, the operating orientation is horizontal or substantially horizontal. In other preferred embodiments, the operating orientation is vertical or substantially vertical. As used herein, a horizontal orientation of a frame essentially means that its two largest dimensions lie on a horizontal plane, while its smallest dimension lies on a vertical plane. Conversely, a vertical orientation of a frame is given when its two largest dimensions lie on a vertical plane and its smallest dimension lies on a horizontal plane. The modifier "substantially" takes into account that a frame may typically have a somewhat flat or planar overall shape, but is not completely flat or planar.

[0045] The frame, particularly one having a vertical operating orientation, may further comprise one or more means for securing or holding it in its proper position and orientation. For example, the frame may include one or more through-holes adapted to mate with protrusions or hooks provided by a device configured to operate the frame or a kit comprising the frame, as defined herein.

[0046] Not only the frame may have an operational orientation. As mentioned above, orientation may also be important for containers, such as flexible substrate containers. In some further preferred embodiments, the static mixing device also has an operational orientation. The frame may advantageously be adapted to ensure that the static mixing device can only be mounted in the proper orientation, i.e., the desired orientation. In particular, the means for holding the static mixing device may be adapted to hold the static mixing device only when the static mixing device has the desired orientation. To avoid confusion, the desired orientation of the static mixing device is defined relative to the frame. The desired orientation of the static mixing device is the same as its operational orientation, provided that the frame is also in that operational orientation.

[0047] In some related embodiments, the desired orientation of the static mixing device is such that when the frame is in its operating orientation, the flow of liquid from the static mixing device through its outlet port occurs in an anti-gravity direction. Such upward or anti-gravity flow of liquid in the static mixing device has been found to reduce undesirable mixing effects such as bubble formation.

[0048] As mentioned above, at least partial anti-gravity flow may be desirable in other portions of the fluid flow path, such as between the substrate container and the static mixing device. In some related embodiments, the first and second sealable regions or cavities and the means for holding the static mixing device are arranged so that, in the operating orientation of the frame, the outlet ports of the flexible substrate container are positioned lower relative to the positions of the respective inlet ports of the static mixing device. Such a configuration requires at least some upward flow of fluid. In this context, we assume that not only is the frame itself in the operating orientation, but that the substrate container and static mixing device are fixed in the correct orientation.

[0049] In many processes requiring the mixing of two liquid substrates, the optimal ratio of the substrates to one another is different from 1:1. More typically, one of the substrates must be provided in a significantly higher amount and / or at a higher rate than the other. Therefore, the two substrate containers may differ in size, and thus, in some embodiments, the first and second sealable areas or cavities differ in size. In this context, the size of the sealable area or cavity primarily refers to the size of the flexible container it can hold. Furthermore, the difference in size of the sealable area or cavity is reflected in the difference in the volume of the sealable area or cavity, or the pressure chamber formed by the sealable area, when covered by the first counterpiece, as further described below, which requires a difference in at least one dimension of the sealable area. In some embodiments, the volume of one of the two cavities or pressure chambers is at least about 50% larger than the volume of the other cavity or pressure chamber relative to the volume of the smaller cavity or pressure chamber. Also preferred are embodiments in which both the length (ie, largest dimension) and width (ie, second largest dimension) of one of the two cavities or sealable areas are greater than the length and width of the other cavity or sealable area.

[0050] As previously mentioned, the frame is particularly useful for pump-free aseptic preparation of sterile liquid products by mixing or reacting two fluid substrates in small batches, with the fluids being forced through the mixing device by pressure. The pressure can be provided, for example, by pressurized gas. Such pressure-driven systems are described, for example, in co-pending European Patent Application No. 21206216 or International Publication No. WO 2023 / 079039, the entire disclosures of which are incorporated herein by reference.

[0051] When pressurized gas is used to force fluid from a flexible substrate container through a conduit and into a static mixing device, the sealable area or cavity in which the container is held must be tightly closed to form a pressurizable chamber (e.g., by applying a lid or another type of counterpiece to cover the sealable area or cavity), and the pressurized gas must be introduced into these chambers in a controlled manner.

[0052] To this end, in some embodiments, the frame or its tray portion is provided with a first through-hole disposed in a first sealable area and a second through-hole disposed in a second sealable area. Again, the sealable areas may be provided as cavities. As used herein, a through-hole is to be understood as any opening in a sealable area, regardless of the size and shape of the through-hole, that allows the sealable area to be pressurized with gas from outside the frame when the sealable area is otherwise covered and sealed. For example, the through-hole may have dimensions not significantly smaller than the dimensions of the sealable area, as seen, for example, in FIG. 3.

[0053] Various options exist for the spatial arrangement of the first and second sealable regions or cavities relative to one another. In some embodiments, the first and second sealable regions or cavities are arranged to hold the flexible substrate containers such that the outlet port of the first flexible substrate container and the outlet port of the second flexible substrate container are coaxially opposed. In some further embodiments, the outlet port of the first flexible substrate container and the outlet port of the second flexible substrate container are coaxially opposed at an angle of about 45° relative to the central longitudinal axis of the frame.

[0054] In some embodiments, the first and second sealable regions are arranged to hold the flexible substrate containers such that the outlet port of the first flexible substrate container and the outlet port of the second flexible substrate container are coaxial and not opposed. In one embodiment, the first and second sealable regions or cavities are arranged to hold the flexible substrate containers such that when the frame is in its operational orientation, liquid flow from each flexible substrate container through its outlet port occurs in an anti-gravity direction. These embodiments are particularly advantageous when the operational orientation of the frame is vertical.

[0055] The frame may also be adapted to hold the flexible product container only in a specific orientation. This can be achieved, for example, by the configuration of the means for holding the container. In some embodiments, the operational orientation of the frame is vertical and is adapted to hold the product container vertically, and its inlet port for receiving liquid from the static mixer is either horizontal or vertical, such that the inflow of liquid from the static mixer through the inlet port occurs in an anti-gravity direction. Such a vertical orientation of the container and inlet port may be advantageous, for example, when the liquid received from the static mixer is intended to be diluted with a liquid diluent provided as a prefill in the product container to promote mixing of the respective liquids on the product container. A horizontal orientation of the inlet port may be advantageous when in-line or subsequent dilution of the liquid received in the product container from the static mixer is intended. In this context, in-line means that the liquid diluent is supplied to the product container while the container also receives liquid from the static mixer, and the liquid diluent is then supplied after the product container no longer receives any liquid from the static mixer.

[0056] As mentioned above, it may be desirable to minimize the length of at least those conduits located between the substrate containers and the static mixing device. Additionally, it has been found advantageous to house these conduits within the same sealable area or cavity as their respective substrate containers. An advantage of such an embodiment is that, when pressurized gas is used as the driving force for the liquid, the conduits are not subjected to large pressure differentials during operation because they are located within the same pressure zone as the flexible substrate containers to which they are connected. A further advantage is that high-pressure seals for the conduits are not required; if the conduits are housed within a sealable area or cavity, the seal may be against the mixing device, which is mechanically more stable than the conduits. Thus, in some embodiments, a first sealable area or cavity is shaped to at least partially house a conduit for fluidly connecting the outlet port of the first substrate container with the first inlet port of the static mixing device. In a related embodiment, the second sealable area or cavity is shaped to at least partially accommodate a conduit for fluidly connecting the outlet port of the second substrate container with the second inlet port of the static mixing device, or both the first and second sealable areas or cavities are shaped accordingly.

[0057] The frame may further contain one or more means for holding conduits for fluidly connecting the outlet ports of the static mixing device with the inlet ports of the waste container and the product container. Again, such means may optionally be molded as recesses or cavities in the frame or tray portion of the frame into which the conduits can be at least partially inserted. When used in the context of a holding means, recesses and cavities may be used interchangeably. The method of holding the conduits in the cavities is particularly suitable for short conduits. As mentioned above, it may be desirable to keep the conduits short so as to have little dead fluid space, which can reduce product yield, especially when producing small product batches. Alternatively or additionally, other holders, such as clamps, clips, sleeves, etc., may be provided.

[0058] In some embodiments, the frame further comprises one or more means for holding valves that can be positioned within a conduit to fluidly connect the outlet port of the static mixing device with the inlet port of a waste container or a product container. For example, one or two valves can be envisioned to control the flow of fluid from the static mixing device to the waste container and the product container. Such valves can also help prevent backflow from the product container during or at the end of a batch process. For example, a single diverter valve can be used to initially direct liquid into a conduit connected to the waste container, and once a stable mixing process is achieved, the diverter valve can direct the fluid to the product container. Alternatively, two valves, such as pinch valves or other stopcock valves, can be used for this function, with the first valve positioned in the conduit or fluid path between the static mixing device and the waste container and the second valve positioned between the static mixing device and the product container. Again, a holder or holding means for such a valve (or valve, as the case may be) can be configured as a recess or cavity in the frame or tray portion of the frame into which the valve is received and held in place. Alternatively, one or more rims, clamps, clips, or other structures may be used for this purpose, or even a combination of both.

[0059] In some related embodiments, the frame is adapted to hold a Y-piece or T-piece, and the Y-piece or T-piece is positioned within a conduit fluidly connecting the outlet port of the static mixing device with the inlet ports of the waste container and the product container, such that the inlet of the Y-piece or T-piece is fluidly connected with the outlet port of the static mixing device, a first outlet of the Y-piece or T-piece is fluidly connected with the inlet port of the waste container, and a second outlet of the Y-piece or T-piece is fluidly connected with the inlet port of the product container.

[0060] In some further related embodiments, one-way stopcock valves may be located downstream of the Y-piece or T-piece, i.e., in the conduit section fluidly connecting the first outlet of the Y-piece or T-piece with the inlet port of the waste container, and / or in the conduit section fluidly connecting the second outlet of the Y-piece or T-piece with the inlet port of the product container. Preferably, one-way stopcock valves are located at both of these locations. In this context, a one-way stopcock valve should be understood as a one-way valve that has an open state and a closed state and does not have a pressure reduction or flow regulation function. Preferably, the one-way stopcock valve can be operated to switch rapidly from its closed state to its open state, particularly in less than 1 second, or even less than 0.5 seconds.

[0061] The one-way stopcock valve, or preferably each one-way stopcock valve, may comprise means for mechanically operating the valve, said means preferably oriented relative to or adapted for operation from a direction corresponding to the rear face of the frame. Such an arrangement allows the valve to be operated by an automated operator device located behind the frame as viewed from the front when the frame is inserted into an apparatus, for example, as described in more detail below. In one embodiment, the frame is adapted for or comprises at least one means for retaining or securing the valve to the frame.

[0062] In some further embodiments, a check valve is disposed in the conduit or conduit section, fluidly connecting the first outlet of the Y-piece or T-piece with the inlet port of the waste container. If the conduit or conduit section also includes a one-way stopcock valve as described above, the check valve is positioned downstream of the one-way stopcock valve. This check valve can be advantageous when it is intended to remove residual gas or air from the flexible substrate container before initiating the mixing process. If the substrate container is oriented such that its respective outlet port is vertically oriented to allow fluid to exit in an anti-gravity direction, the container may be degassed by slightly squeezing the container while the stopcock valve upstream of the waste container is in its open state, thereby pushing the air or gas toward the waste container. The check valve then prevents any backflow of gas or air when the substrate container is no longer squeezed for degassing. Examples of potentially suitable check valves include duckbill valves, ball valves, swing valves, piston valves, butterfly valves, and tilting disk valves. In some preferred embodiments, the check valve is a duckbill valve.

[0063] In some further embodiments, the means for holding the static mixing device itself is molded as a recess or cavity in the frame or the tray portion of the frame. As noted above, according to one preferred embodiment of this aspect of the invention, the frame is positioned so that, in its operational orientation, liquid flow through the outlet port of the static mixing device is in an upward or anti-gravity direction. Thus, the cavity for holding the mixing device may be shaped and oriented to allow insertion of the mixing device only in its desired orientation, which is the operational orientation when the frame itself is also in that operational orientation. One or more additional means for holding the mixing device, such as a clamp, may also be used.

[0064] According to some further preferred embodiments, the frame or its tray portion includes a circumferential gasket for each of the first and second sealable areas or cavities. The use of gaskets to seal the first and second sealable areas or cavities against the counterpiece is generally preferred, particularly when holding the respective substrate containers and when the cavities are intended to be pressurized when covered by such a counterpiece. While the gaskets may be pre-positioned on either the frame or its tray portion or the counterpiece, it is currently preferred to provide the gaskets together with the frame, i.e., to provide a frame that includes a gasket. Optionally, one gasket may be provided that is molded as a circumferential seal around the first sealable area or cavity and around the second sealable area or cavity. In further embodiments, each of the first and second sealable areas or cavities of the frame is independently surrounded by a first circumferential gasket positioned on the front surface of the sealable area (e.g., the side facing the user relative to the frame) and a second circumferential gasket positioned on the rear surface of the frame around the sealable area. As understood herein, the circumferential gaskets may be independently fitted to the first and second sealable areas or cavities and / or the front or rear surfaces of the sealable areas. Furthermore, one or more gaskets sealing the first and second sealable areas or cavities may also be shaped such that a gasket portion also seals the respective sealable area or cavity against a portion of the static mixing device, for example, against one of the inlet ports. Alternatively, the inlet port of the static mixing device may be sealed against a hollow tubular structure provided on the frame, such as a circumferential rim that defines the sealable area or cavity.

[0065] As previously mentioned, it is often desirable to minimize dead volumes, which can lead to fluid loss and reduced product yield. Therefore, it is preferable to use relatively short conduits between various containers and the static mixing device. To this end, according to some further preferred embodiments, the distance between the first sealable area or cavity and the second sealable area or cavity is relatively short, e.g., less than half the length of each of the sealable areas or cavities. In this context, the length of a sealable area or cavity is its largest dimension, and the distance should be understood as the shortest distance between the locus on the contour of the first sealable area or cavity and the locus on the contour of the second sealable area or cavity. Keeping in mind that the dimensions of the two sealable areas or cavities may differ from each other, the distance between the two sealable areas or cavities must be smaller than the length of the smaller of the sealable areas or cavities.

[0066] According to a preferred use of the frame, some further preferred embodiments provide that the frame is adapted to be insertable in its operative orientation into an apparatus for aseptically mixing two fluids, the apparatus comprising a counterpiece for sealably covering the first and second sealable areas, and means for applying pressure to the first and second flexible substrate containers when secured to or inserted into the first and second sealable areas. With regard to an apparatus which is part of the present invention and relates to one of its further aspects, reference is made to the respective section of the detailed description below.

[0067] As will be understood by those skilled in the art, the present invention encompasses the frame itself, as described above, and frames with certain components assembled. In other words, in some embodiments, the frame holds first and second substrate containers, a flexible waste container, a flexible product container, a static mixing device, and conduits for fluidly connecting the outlet port of the first substrate container with the first inlet port of the static mixing device, the outlet port of the second substrate container with the second inlet port of the static mixing device, and the outlet port of the static mixing device with the inlet ports of the waste container and the product container. Furthermore, in these embodiments, the conduits may actually be fitted to fluidly connect the outlet port of the first substrate container with the first inlet port of the static mixing device, the outlet port of the second substrate container with the second inlet port of the static mixing device, and the outlet port of the static mixing device with the inlet ports of the waste container and the product container. Thus, the assembled frame can be immediately used, for example, by filling the flexible substrate containers with the first and second substrates, unless pre-filled substrate containers are assembled in the frame.

[0068] In some preferred embodiments, the frame is adapted to be insertable into an apparatus for aseptically mixing two fluids as described above, and the pressure is applied by pressurized gas contacting the outer surfaces of the first flexible substrate container and the second flexible substrate container, the pressurized gas being provided to said surfaces through a first through-hole disposed in the first sealable area and a second through-hole disposed in the second sealable area. In other words, the apparatus is configured to externally pressurize the flexible substrate containers through the through-holes disposed in the sealable areas to pump fluids from the substrate containers toward the static mixing device.

[0069] For this purpose, as defined above, the rear face of the frame is preferably adapted to be sealed against the second counterpiece by circumferential gaskets that individually surround the first and second through-holes. The circumferential gaskets may be provided by the frame and disposed on its rear face, or they may be provided by and disposed on a surface of the second counterpiece that is configured to contact the frame.

[0070] In some embodiments, the first and second counter pieces are hinged or connected to each other. Particularly when the frame has a vertical operating orientation, the frame may further comprise one or more means or structures configured to hold or reversibly attach the frame to the second counter piece in a predetermined position. For example, the frame may exhibit two or more through-holes that mate with corresponding hooks or protrusions provided on the contact surface of the second counter piece. In this manner, the frame fitted with the flexible container, static mixing device, conduit, and any other components as described above may be attached to the second counter piece, and the first counter piece may then be placed against the second counter piece to surround the frame. This results in the sealable area in the frame being in sealing contact with the first counter piece, and the rear surface of the frame being in sealing contact with the second counter piece.

[0071] In a further aspect, the present invention relates to a kit comprising the frame described herein. The kit further comprises any one or combination of the following components: (a) a first and / or second flexible substrate container, (b) a flexible waste container, (c) a flexible product container, (d) a static mixing device, and / or (e) one or more conduits for fluidly connecting the outlet port of the first substrate container with the first inlet port of the static mixing device, the outlet port of the second substrate container with the second inlet port of the static mixing device, and the outlet port of the static mixing device with the inlet ports of the flexible waste container and the flexible product container. According to one preferred embodiment, the kit comprises all of components (a) through (e), including both the first and second flexible substrate containers and all conduits for the designated fluid connections. In a related embodiment, the kit also comprises components that can be used in conjunction with or disposed within conduits, such as conduits for connecting the outlet port of the static mixing device with the inlet ports of the flexible waste container and the flexible product container, respectively. For example, the kit may include any one or combination of a T-piece, a Y-piece, or a valve, or any other component described herein, that may be useful in assembling the kit, operating the assembled frame or assembled kit, or performing any of the methods and processes described herein.

[0072] In some preferred embodiments, the flexible substrate container, flexible waste container, and flexible product container are the containers disclosed herein above. In particular, the flexible product container may include at least one inlet port and / or at least one outlet port to which a flexible tube including a sterile disconnect is fluidly connected. Furthermore, the flexible tube fluidly connected to the at least one outlet port of the product container may have a downstream end fluidly connected to a sampling tube, and the sampling tube may have a downstream end fluidly connected to a sterile filter.

[0073] In some embodiments, the kit is provided with an empty substrate container, allowing the user to fill the substrate container with the desired fluid material. In other embodiments, the kit is provided with first and second containers, preferably already filled with liquid substrate. If so, the liquid substrate contained in the first flexible substrate container, also referred to as the first substrate, is preferably different from the liquid substrate contained in the second flexible substrate container, also referred to as the second substrate. The first and second substrates are typically selected to form a liquid product, such as a liquid pharmaceutical composition for injectable use, when mixed in a static mixing device. As described above, the frame and, therefore, the kit comprising the frame, are particularly useful for preparing sterile liquid products in small batches using a simplified, standardized, and substantially error-proof process that relies on disposable components that do not need to be cleaned after batch preparation. As a result, the preparation of small batches of sterile injectable products can be highly efficient based on the present invention.

[0074] Examples of types of sterile products that can be prepared by mixing two liquid substrates are pharmaceutical compositions containing lipid nanoparticles (LNPs), liposomes, or similar colloidal carriers of active ingredients. Some modern mRNA-based vaccines are based on LNPs, which can be prepared by mixing an organic solution of lipids with an aqueous solution of mRNA. Thus, in some embodiments, a first flexible container is filled with a first substrate, preferably a sterile organic solution of one or more lipids capable of forming LNPs, and a second flexible container is filled with a second substrate, preferably a sterile aqueous solution of mRNA, such as mRNA capable of expressing an antigen.

[0075] As previously mentioned, a kit according to one embodiment of the present invention may include some or all of the components necessary to assemble a frame ready for use. Kits representing any partial selection of components configured for use with the frame are also within the scope of the present invention. For example, a kit may consist only of empty or pre-filled first and second flexible substrate containers configured for the preparation of a particular product.

[0076] In some embodiments, the flexible product container is partially pre-filled with a third substrate. Such a third substrate may be used as an additional component of the final product that is desirable to add only after the mixing process, i.e., after the mixing of the first and second substrates in a static mixing device. For example, the third substrate may be a diluent, such as a sterile aqueous diluent or a sterile aqueous buffer. For example, the formation of mRNA-loaded LNPs by mixing an organic lipid solution with an aqueous mRNA solution may be achieved better and more efficiently at a first pH, but the long-term stability of the final product is higher at a second pH. In such cases, it may be useful to partially pre-fill the flexible product container with a sterile aqueous buffer to buffer the final product at the second pH.

[0077] Again, the flexible product container may be provided in empty form or partially pre-filled, and may be provided together with the frame and / or any other components required to assemble the frame so that it is ready for use.

[0078] The static mixing device provided for use with the frame or adapted to be held by the frame, e.g., as part of a kit, may be selected from any static mixing device capable of aseptically mixing two liquids. In some embodiments, the static mixing device comprises or consists of a T-piece mixer, a Y-piece mixer, a vortex mixer, a baffle-based static mixer, a microfluidic mixing device, a multi-inlet vortex mixer, or a jet impingement reactor. In some preferred embodiments, the static mixing device is a jet impingement reactor.

[0079] Jet impingement reactors are fluid reactors for mixing fluids or generating particulate fluids through collision. They can be used, for example, to produce nanoparticle fluids incorporating poorly water-soluble active ingredients. The function of these reactors is based on the use of two fluid streams (at least one of which typically contains the active ingredient) injected into the reactor cavity and colliding in a turbulent mixing zone, thereby creating nanoparticles. One of the main principles used in conjunction with jet impingement reactors is solvent / non-solvent precipitation, in which a first fluid containing the active ingredient dissolved in a suitable solvent is contacted with a non-solvent or anti-solvent under defined conditions to precipitate nanoparticles containing the active ingredient. If one of the solvents contains lipids, lipid nanoparticles can be produced with the aid of a jet impingement reactor, which can then be loaded with a biologically active compound, for example, by pH shift.

[0080] A jet impingement reactor comprises a reaction chamber with two fluid inlets with nozzles that allow two fluids to be injected into the reaction chamber, typically at pressures greater than ambient pressure. Through the first and second fluid inlets, the two streams are injected so that they meet inside the reaction chamber, forming a collision or mixing zone. An outlet is also provided for obtaining the resulting product.

[0081] An example of a jet impingement reactor is the microjet reactor disclosed in EP 1165224. Such a microjet reactor has at least two nozzles or pinholes positioned opposite each other, each with an associated pump and supply line for directing liquids toward a common impingement point in a reaction chamber enclosed by a reactor housing. The reaction chamber comprises two bores, which intersect with each other and result in a small cavity where the two fluids impinge, possibly without contacting the walls of this cavity. One of the bores contains two fluid inlets, while the second bore contains an additional opening in the reactor housing through which a gas, evaporating liquid, cooling liquid, or cooling gas can be introduced to maintain or cool the gas atmosphere in the reaction chamber. The other end of the second bore is provided with an additional opening for removing the resulting product and excess gas from the reactor. Solvent / non-solvent precipitation in such a microjet reactor results in a dispersion of precipitated particles. The reactor can also be adapted to use a third fluid, such as an external source of gas or cooling liquid.

[0082] WO 2018 / 234217 discloses another jet impingement reactor having a housing enclosing a reaction chamber and collinearly oriented first and second fluid nozzles. The second nozzle is located directly opposite the first fluid nozzle in terms of the nozzle's jet direction. The nozzles reach the reaction chamber and form a disk-shaped impingement zone with each other. This reactor type has at least one rinse fluid inlet located on the side of the first fluid nozzle and at least one product outlet located on the side of the second fluid nozzle, making it suitable for continuous production of fine particle fluids. Furthermore, the rinse fluid conducting structure is designed as parallel channels on the side of the first fluid nozzle, which produce a rinse fluid flow directed toward the jet direction of the first fluid nozzle and direct the rinse fluid toward the impingement disk, causing a slight deformation of the impingement disk. This transports particles generated in the reactor away from the impingement zone. Therefore, the production process when carried out in the reactor disclosed in WO 2018 / 234217 relies on the presence of a rinse fluid conducting structure and a rinse fluid.

[0083] In some preferred embodiments, the jet impingement reactor used in the context of the present invention is the device described in International Publication No. WO 2023 / 025736, the entire disclosure of which is incorporated herein by reference. The jet impingement reactor comprises a reaction chamber defined by the inner surface of a reaction chamber wall, the reaction chamber having a substantially spheroidal overall shape. The reaction chamber further comprises first and second fluid inlets, the first and second fluid inlets being positioned at opposite positions on a first central axis of the reaction chamber so as to face each other, each of the first and second fluid inlets comprising a nozzle, and a fluid outlet positioned at a third position, the third position being located on a second central axis of the chamber, the second central axis being perpendicular to the first central axis. Furthermore, the distance between the nozzle of the first fluid inlet and the nozzle of the second fluid inlet is equal to or less than the diameter of the reaction chamber along the first central axis.

[0084] In some preferred embodiments, each nozzle has a downstream end that is substantially aligned with the inner surface of the chamber wall. Furthermore, the reaction chamber preferably does not include any additional inlet or outlet openings. According to a further preferred embodiment, each of the first and second fluid inlets is provided by a fluid inlet connector having an upstream end, a downstream end that holds a nozzle of the first or second fluid inlet, and a fluid conduit for conducting fluid from the upstream end to the downstream end, and the downstream end of each fluid inlet connector is reversibly insertable into the chamber wall to provide the first and second fluid inlets.

[0085] The reactor is useful for mixing two fluids by a method comprising the steps of providing a jet impingement reactor according to the present invention, directing a first fluid stream into a reaction chamber through said first fluid inlet, and directing a second fluid stream into the reaction chamber through a second fluid inlet so as to collide with the first fluid stream at an angle of about 180°.

[0086] Furthermore, in some preferred embodiments, the orifice of the first nozzle is larger than the orifice of the second nozzle, and / or the flow rate of the first fluid is larger than the flow rate of the second fluid, and the pressures of the first and second fluids may be adapted so that the first and second fluid streams have substantially the same kinetic energy when entering the reaction chamber.

[0087] The jet impingement reactor described in WO 2023 / 025736 may be made by injection molding. For example, the jet impingement reactor, or at least the reactor wall, may be made from a thermoplastic polymer by injection molding, and a prefabricated inlet nozzle made of a hard, non-thermoplastic material such as metal, glass, or ceramic may be inserted into the mold during the injection molding process, or mechanical or laser drilling may be used to manufacture nozzles on both sides of the reactor.

[0088] Another jet impingement reactor particularly useful in practicing the present invention is the device described in co-pending European Patent Application No. 22195145.2, the entire disclosure of which is incorporated herein by reference. The jet impingement reactor comprises a housing made of a polymeric material, the housing enclosing a reaction chamber, said chamber having a substantially spherical shape, the spherical shape interrupted only by at least first and second fluid inlets, the first and second fluid inlets being positioned at opposite positions on a first central axis of the reaction chamber so as to face each other, each of the first and second fluid inlets being provided by a nozzle, and a fluid outlet positioned at a position located on a second central axis of the chamber, the second central axis being perpendicular to the first central axis. the reactor further comprises first, second, and third fluid conduits, the first and second fluid conduits arranged to direct the first fluid to the first fluid inlet and the second fluid to the second fluid inlet, the third fluid conduit arranged to direct the third fluid downstream from the fluid outlet, the third fluid being formed by mixing or reacting the first and second fluids in the reaction chamber, the housing comprising at least two pieces secured together, the first piece of which includes at least a majority of the first or second fluid conduits, and the second piece of which comprises at least one of the nozzles.

[0089] The jet impingement reactor may also be prepared by a method comprising injection molding the first and / or second pieces of the housing. It is described as being particularly useful in the aseptic manufacture of sterile liquid pharmaceutical compositions.

[0090] According to a further preferred embodiment, the kit comprises a jet impingement reactor having a reaction chamber having a substantially spherical shape, the spherical shape being interrupted by first and second fluid inlets disposed at opposing positions on a first central axis of the reaction chamber such that the first and second fluid inlets face each other, each of the first and second fluid inlets being provided by a nozzle, and a fluid outlet disposed at a position located on the second central axis of the chamber, the second central axis being perpendicular to the first central axis. The reactor further comprises first, second, and third fluid conduits, the first and second fluid conduits being disposed to direct the first fluid to the first fluid inlet and the second fluid to the second fluid inlet, and the third fluid conduit being disposed to direct the third fluid downstream from the fluid outlet, the third fluid being formed by mixing or reacting the first and second fluids in the reaction chamber. Further, the reactor comprises at least two pieces fixed to one another, a first piece of which is made of a polymeric material and comprises at least a portion of the first or second fluid conduit and at least a hemispherical portion of the reaction chamber, and a second piece of which is at least partially insertable into the first piece and comprises a fluid outlet. Such reactors are described in co-pending European Patent Application No. 23163257.1 or PCT / EP2023 / 075054, the entire disclosures of which are incorporated herein by reference.

[0091] According to some further preferred embodiments, each of the components or parts of the kit is sterile or sterilizable by heat, irradiation, or ethylene oxide. For example, the flexible containers, i.e., substrate container, waste container, and / or product container, may be made of materials similar to infusion bags or mixing bags for parenteral solutions, such as ethylene-vinyl acetate copolymer (EVA), which can be autoclaved or steam sterilized (e.g., in steam at 121°C for 15 minutes), gamma sterilized (e.g., at 25-40 kGy), or sterilized with ethylene oxide. Preferably, the static mixing device, like the conduit, is provided in a sterile form or is sterilizable, even if made substantially from a polymeric material.

[0092] It should be noted that the present invention is particularly useful for carrying out aseptic mixing processes using only sterile starting materials and pre-sterilized containers and product-contacting equipment parts, in which case sterilization of the final product may not be necessary.

[0093] The kit may include any additional features described in the context of the frame. In particular, as one skilled in the art will appreciate, any preferences disclosed above in the context of the frame should also apply to the kit.

[0094] In a further aspect, the present invention relates to a flexible container adapted for use as a flexible substrate container, flexible waste container, or flexible product container as identified hereinabove. The container has an interior space for holding a fluid material surrounded by a flexible front wall and a flexible rear wall, each wall being made of a polymeric material. The container further comprises at least one inlet or outlet port for allowing fluid communication with the interior space. The flexible front wall and the flexible rear wall are connected to each other to form a sealed edge that substantially encloses the interior space. The edge includes four corner regions such that the interior space has an overall square or rectangular shape when empty. The container is further characterized by having at least two through-holes provided in the sealed edge, a first through-hole at or near a first corner region of the edge and a second through-hole at or near a second corner region of the edge, the second corner region adjacent to the first corner region.

[0095] The flexible front wall and the flexible rear wall may be formed from two sheets of similarly sized and shaped flexible polymeric material that are secured (e.g., welded) together to form a sealed (e.g., welded) edge that substantially encloses the interior space. The two sheets may have an essentially square or rectangular overall shape, i.e., four corners, which also translates to the essentially square or rectangular overall shape of the interior space when the interior space is empty. In this context, corners should be understood to include rounded corners. In some embodiments, the corners of the container and / or interior space are rounded corners. Also, in this context, the terms "essentially," "substantially," and "entirely" should be understood as modifiers that allow for minor deviations. For example, the sealed edge may enclose the entire interior space except where the inlet or outlet ports are located, and the square or rectangular overall shape may not represent a perfect square or rectangular shape. Furthermore, the phrases "near a first corner region" or "near a second corner region" should be understood to refer to a location closer to the respective corner than any central axis of the container along any of its three dimensions.

[0096] The at least two through-holes allow the container to be held by or secured to complementary retaining means, such as pins. As discussed above in connection with a sealable area, such as a cavity in a frame or a tray portion of a frame, such means for retaining a flexible container may be provided in the cavity. In addition to their fastening function, they may also contribute to assembly error prevention in that a specific spatial arrangement of the through-holes and complementary retaining means (e.g., pins) can be used to match only the correct or designated container and its desired orientation. In other words, the positions of the first and second through-holes correspond to the positions of corresponding retaining means provided in the sealable area or cavity of the frame or tray portion configured to hold the flexible container. In some preferred embodiments, each flexible container has at least three through-holes. Containers with four through-holes are also preferred.

[0097] In some embodiments, the flexible container comprises at least one inlet port and / or at least one outlet port. Preferably, flexible tubing is fluidly connected to the inlet port and / or outlet port. The tubing may comprise a sterile disconnector. Optionally, each inlet and outlet port of the flexible container comprises a sterile disconnector. In this context, a sterile disconnector refers to an easily severable transfer conduit or conduit segment, such as those described in WO 2010 / 008396, or another conduit with the same or similar functionality. Such embodiments are particularly advantageous as flexible product containers, as they facilitate rapid removal of the product after the mixing process while minimizing the risk of microbiological product contamination.

[0098] In some further embodiments of the flexible container, the flexible tubing fluidly connected to the at least one outlet port has a downstream end fluidly connected to a sampling tube, which has a downstream end fluidly connected to a sterile filter. Again, such embodiments are particularly advantageous for flexible product containers as they allow for easy sample removal from the product container without risk of product contamination.

[0099] Additionally, in some further embodiments, the flexible container includes an identification tag, such as an RFID tag. As noted above, an identification tag, such as an RFID tag, may also be featured on the frame itself, the static mixing device, or any other component of the kit specified above.

[0100] In a further aspect, the present invention relates to an apparatus for operating a frame in its assembled state. In some embodiments, such an apparatus functions as a filling station. More specifically, such an apparatus may be adapted to receive a frame as described above in its operational orientation, with the first and second flexible substrate containers empty and held in the first and second sealable areas. The apparatus further comprises means for aseptically filling the first and / or second substrate into the first and / or second flexible substrate containers, respectively. In these embodiments, the frame is fully assembled, i.e., holds the first and second substrate containers, a flexible waste container, a flexible product container, a static mixing device, and conduits for fluidly connecting the outlet port of the first substrate container with the first inlet port of the static mixing device, the outlet port of the second substrate container with the second inlet port of the static mixing device, and the outlet port of the static mixing device with the inlet ports of the waste container and the product container. Furthermore, the conduits are fitted to actually provide the respective fluid connections. In some embodiments, the apparatus further comprises means for partially filling the flexible product container with a third substrate, such as a diluent or aqueous buffer, as further described above.

[0101] In some further preferred embodiments, such an apparatus is configured to function as a mixing station. Specifically, such an apparatus may be adapted to receive a frame in its operational orientation, which assembles and holds first and second substrate containers, a flexible waste container, a flexible product container, a static mixing device, and conduits fluidly connecting the outlet port of the first substrate container with the first inlet port of the static mixing device, the outlet port of the second substrate container with the second inlet port of the static mixing device, and the outlet port of the static mixing device with the inlet ports of the waste container and the product container. In these embodiments, the first and second flexible substrate containers are pre-filled, i.e., contain the first and second substrates, respectively. Furthermore, the apparatus further comprises means for forcing the first substrate and the second substrate from the first and second flexible substrate containers to flow into the static mixing device to mix and form a liquid product. As one of ordinary skill in the art would understand based on the above disclosure, the liquid product flows from the static mixing device through a conduit to a flexible waste container and / or a flexible product container, depending on the settings of valves that may be disposed in the conduits connecting the outlet port of the static mixing device with the waste container and the product container. Also, in these embodiments, optionally, the product container may be partially filled with a third substrate, as previously described.

[0102] The means for forcing the first and second substrates from the first and second flexible substrate containers into the static mixing device may comprise one or more pumps. In some preferred embodiments, the means is adapted to apply pressure to the first and second flexible substrate containers to squeeze out the substrates. Particularly preferred embodiments include those in which the means for forcing the substrates from the first and second flexible substrate containers into the static mixing device comprises pressurized gas. One example of a pressurized gas is pressurized air. In other words, such preferred embodiments provide an apparatus in which pressure is applied by pressurized gas contacting the outer surfaces of the first and second flexible substrate containers, and the pressurized gas is provided to said surfaces through a first through-hole disposed in the first sealable region and a second through-hole disposed in the second sealable region.

[0103] The device may further include a counterpiece for the frame that individually seals and covers the first and second sealable areas, thereby allowing the sealable areas to hold the first and second substrate containers. Each sealable area and the corresponding cover provided by the first counterpiece together form a pressurizable chamber. The chamber may be pressurized by a pressurized gas that can enter the chamber through a through-hole provided in the sealable area, as described above. Preferably, the chambers are individually sealed, i.e., also sealed to each other.

[0104] In some further preferred embodiments, the device also comprises a second counterpiece, as described above. Furthermore, the first counterpiece and the second counterpiece may be hingedly connected to each other.

[0105] In some preferred embodiments, the pressurized gas is provided by a pressure reservoir chamber. Such a pressure reservoir chamber is adapted to hold a specific volume of pressurized gas and, when fluidly connected to the sealed chamber containing the flexible substrate container, can immediately pressurize them. As mentioned above, the chamber holding the substrate container, also referred to as the substrate chamber, should preferably be individually sealed. Therefore, it is also preferable to provide a separate pressure reservoir chamber for each substrate chamber. Preferably, any piping connecting the pressure reservoir chamber to the substrate chamber should be as short as possible and have a diameter large enough to allow rapid pressure equilibration. The use of gas pressure to push the liquid substrate through the static mixing device and the use of a pressure reservoir chamber to provide such gas pressure are also described in detail in co-pending European Patent Application No. 21206216.0 or International Publication No. WO 2023 / 079039, the entire disclosures of which are incorporated herein by reference.

[0106] In yet another aspect, the present invention relates to methods, particularly methods for mixing or reacting two fluid substrates, involving the use of the above-described frame, kit, flexible container, or device. In some preferred embodiments, such methods are carried out aseptically, i.e., using sterile substrates and pre-sterilized containers and product-contacting equipment components.

[0107] In a related aspect, the present disclosure also relates to a process for preparing an assembled frame or an assembled kit according to any one or combination of the respective embodiments described herein for use in a method of mixing or reacting two liquid substrates, the process comprising at least: a) filling first and second flexible substrate containers with first and second liquid substrates, the first and second liquid substrates preferably being different; b) optionally filling the flexible product container with a third liquid, such as a buffer; c) removing residual gas or air from the first and second flexible substrate containers; Optionally, any one or all of steps a)-c) are performed aseptically or under sterile or clean room conditions.

[0108] In one embodiment, at least step a) and optionally step b) of filling the vessels are carried out under sterile conditions. In some embodiments, filling of the substrate vessel defined in step a) or optionally the product vessel of step b) may be carried out via an inlet port of said vessel connected to a sterile disconnect as described herein above.

[0109] Steps a) to c) are preferably carried out on a fully assembled frame comprising all components (e.g., of or provided with the kit) necessary to carry out the mixing method. However, in an optional embodiment, the process preferably includes at least the following steps prior to step a) of filling the container and optional step b): first and second flexible substrate containers; -Flexible product container flexible waste containers, static mixing devices, as well as one or more conduits, and optionally components disposed within the conduits, such as a Y-piece and / or a T-piece, for example, but not limited to, fluidly connecting the outlet port of the first substrate container with the first inlet port of the static mixing device, the outlet port of the second substrate container with the second inlet port of the static mixing device, and the outlet port of the static mixing device with the inlet port of the flexible waste container and the inlet port of the flexible product container; or Any of the components of the kits provided herein may include a first step of assembling them onto a frame.

[0110] In a related optional embodiment, a step of sterilizing the assembled kit may also be performed.

[0111] In one embodiment, the frame is prepared for use according to the above process in its operational orientation. For example, any one or combination of steps a)-c) may be performed when the frame is in its operational orientation, which is preferably a vertical operational orientation. In some embodiments, step c) is performed when the frame is placed in an apparatus adapted to receive the assembled frame in its operational orientation. The apparatus, in one embodiment, may function as a mixing station and be configured to operate the frame in its assembled state.

[0112] In alternative embodiments, the frame is prepared for use with respect to steps a)-c) when the frame is at an oblique angle relative to the vertical axis, rather than while in its vertical direction of operation. In some embodiments, any one or combination of steps a), b), or c), but preferably at least step c), is performed when the frame is in an oblique position, e.g., when the upper edge of the frame is positioned at an angle of at least about 25°, 30°, 35°, 40°, or 45°, or at an angle of at least 25-45°, from the vertical axis of its operational orientation.

[0113] As noted above, the present disclosure also provides an apparatus that functions as a filling station. In one embodiment, the apparatus may be adapted to receive a fully assembled frame and may be adapted to hold the frame in an angular position other than its operative orientation, for example, at an angle of 25° to 45° away from the vertical axis as described above, instead of or in addition to holding the frame in its vertical operative orientation.

[0114] Step c) may be performed, for example, by applying positive pressure or negative pressure, e.g., vacuum, to remove residual gas (e.g., inert gas or air) in the flexible container. For example, removal of residual gas may be performed by applying external pressure to the exterior surface of the flexible container in an amount and / or duration sufficient to remove residual gas from the container once the frame is oriented in its operating orientation or another preferred orientation for performing step c), e.g., at an angle as described above. In one embodiment, removal of residual gas from each of the first and second substrate containers is performed sequentially, i.e., not simultaneously. In some embodiments, clamping means, e.g., tube clamps, may be used to prevent premature flow of liquid substrate from one flexible substrate bag into the static mixer while step d) is being performed on the other flexible substrate bag. Clamping means may also be used to prevent fluid flow during, or during any one of, the steps of the above-mentioned process, such as during transport of the assembled frame or assembly of the frame, and / or during removal of residual gas or air from the flexible substrate container, as needed.

[0115] In some embodiments, residual gas is removed and collected in a flexible waste container. In other embodiments, depending on the configuration of the assembled frame, residual gas is removed externally from the assembled frame, rather than to a flexible waste or product container, for example, via a port or portion of a conduit configured in a conduit that is in fluid communication with an inlet port of a product or waste container.

[0116] In some preferred embodiments, the method includes the use of a substrate that, when mixed, results in an aqueous pharmaceutical composition containing colloidal particles, such as nanoparticles, containing an active ingredient. For example, the nanoparticles may represent lipid nanoparticles (LNPs) carrying an active ingredient, such as a nucleic acid (e.g., mRNA) capable of expressing an antigen. In other words, the product obtained by mixing the first and second substrates may be a pharmaceutical composition corresponding to a vaccine.

[0117] Additional liquid compositions that can be prepared by mixing two fluid substrates, preferably under sterile conditions, are generally known to those skilled in the art.

[0118] Moreover, as one skilled in the art will appreciate, various optional or preferred features are described above in the context of frames, kits, and kit components that should also apply to the methods and processes provided by the present invention.

[0119] Detailed Description of the Drawings 1, not drawn to scale, shows a perspective view of a frame (10) according to the present disclosure in an operational orientation that is vertical and parallel to a vertical axis (100). The frame comprises a tray portion (15), which in the illustrated embodiment also forms the entire frame. The frame comprises: (a) a first flexible substrate container (11) and a second flexible substrate container (12), each having an outlet port (91) and a sealed inlet port (92), respectively; (b) a flexible waste container (13) having an inlet port (93); (c) a flexible product container (14) having an inlet port (94) and a resealable outlet port (95); (d) a static mixing device (16) having a first inlet port (96) and a second inlet port (97), and an outlet port (98); and (e) a flexible substrate container (15) having an inlet port (99) and a resealable outlet port (99). ) simultaneously holds a conduit (17a) for fluidly connecting the outlet port (91) of the first substrate container (12) with the first inlet port (96) of the static mixing device, a conduit (17b) for fluidly connecting the outlet port (91) of the second substrate container (12) with the second inlet port (97) of the static mixing device (16), and conduits (17c, 17d) for fluidly connecting the outlet port (98) of the static mixing device (16) with the inlet ports (93, 94) of the product container (14) and the waste container (13). The frame (10) further comprises means (18) for holding valves disposed in said conduits (17c, 17d).

[0120] The illustrated frame (10) comprises cavities (211, 212, 224, 225) shaped to hold each of the containers. The containers (11, 12, 13, 14) are secured in their respective cavities by a plurality of means (26) for securing the containers. As illustrated in this depiction, the means for securing (26) are provided in up to four different locations (not all of which are shown; see also Figures 2, 3, and 4), with securing points located in the peripheral zone or sealing edge of the containers and at or near the four corners of each container. Those skilled in the art will understand that the number of securing points or means is not limited and can vary depending, for example, on the type of means selected for securing the containers, the dimensions of the containers, and possibly the operational orientation of the frame.

[0121] In the illustrated embodiment, the first cavity (211) is shaped to hold a first flexible substrate container (11), a sealed inlet port (92), an outlet port (91), and a conduit (17a) connecting the outlet port (91) of the container with the first inlet port (96) of the static mixing device (16) and the inlet port (96) of the static mixing device. The second cavity (212) is shaped to hold a second flexible substrate container (12), a sealed inlet port (92), an outlet port (91), and a conduit (17b) connecting the outlet port (91) of the second substrate container (12) with the second inlet port (97) of the static mixing device (16), and at least partially with the second inlet port (96) of the static mixing device (16) itself. The first cavity (211) and second cavity (212) each also include a circumferential gasket (19) that is conformally shaped to the respective cavity and that is used to form a seal thereon when an appropriate counterpiece or housing device is applied thereover. The cavity (224) for holding a flexible product container (14) is also shaped to accommodate at least a portion of the inlet port (94) and resealable outlet port (95) of said container. The cavity (225) for holding a flexible waste container (13) is also shaped to accommodate at least a portion of the inlet port (93) of said container (13).

[0122] As shown, the first and second flexible substrate containers (11, 12) and the static mixing device (16) are held in the cavities (211, 212, 224, 225) of the tray portion (15) of the frame (10) in an arrangement such that fluid flow from the first and second flexible substrate containers (11, 12) to the static mixing device (16) occurs at least partially in a direction opposite to gravity, and fluid flow from the static mixing device (16) to the waste container (13) or the product container (14) occurs at least partially in a direction opposite to gravity. Furthermore, the means for holding the first and second cavities (211) and the static mixing device are positioned such that, in the operational orientation of the frame (10), the outlet ports (91) of both flexible substrate containers (11, 12) have lower positions relative to the positions of the respective inlet ports (96, 97) of the static mixing device (16).

[0123] Figure 2, not drawn to scale, shows a front view of the frame (10) of Figure 1 in a vertical operating orientation parallel to a vertical axis (100), with a tray portion (15) that holds the assembly of parts described in Figure 1. As shown, each cavity holding the first and second flexible substrate containers (11, 12) each includes a circumferential gasket (19) around the cavity.

[0124] FIG. 3 shows the same front view of the frame (10) of FIG. 2, not drawn to scale and parallel to the vertical axis (100), but in the operating position without the first and second flexible substrate containers, flexible waste container and flexible product container, static mixing device, and any of the conduits.

[0125] FIG. 4, also not drawn to scale, shows a perspective view of the frame (10) of FIG. 3 in an operational orientation without the assembled first and second flexible substrate containers, flexible waste container and flexible product container, static mixing device, and any associated conduits.

[0126] Both Figures 3 and 4 show a frame (10) comprising a tray portion (15) comprising: a) a first cavity (211) shaped to hold a first flexible substrate container and its outlet and inlet ports, and conduits for fluidly connecting the outlet port of said container with the first inlet port of the static mixer and the first inlet port of the static mixing device; b) a second cavity (212) for holding a second flexible substrate container and its outlet and inlet ports, and conduits for fluidly connecting the outlet port of said container with the second inlet port of the static mixer and the second inlet port of the static mixing device; and c) means (223) for holding a static mixing device, wherein the means (223) is in the form of a recess in the tray portion (15) and is adapted to accommodate at least a portion or partial outer shape of the static mixing device.

[0127] As shown in the embodiment of Figure 3, each first and second cavity (211, 212) is individually provided with a circumferential gasket (19) in said corresponding cavity, with a portion of each gasket in each cavity being provided to be sealable against a portion of the static mixing device and the respective inlet port.

[0128] In the illustrated embodiment, the cavities (211, 212, 224, 225) for holding the first and second flexible substrate containers, for holding the flexible product container, and for holding the flexible waste container each include a through-hole (27), i.e., an opening in the cavity shaped to accommodate the respective container and its associated features. The through-hole (27) of the first cavity (211) is shaped to accommodate the first flexible substrate container and at least a portion of its outlet and inlet ports, as well as conduits for fluidly connecting the outlet port of said container to the first inlet port of the static mixer and the first inlet port of the static mixer. The through-hole (27) of the second cavity (212) is shaped to accommodate the second flexible substrate container, its outlet and inlet ports, as well as at least a portion of the conduits for fluidly connecting the outlet port of said container to the second inlet port of the static mixer. As described herein, the through holes (27) in the first and second cavities (211, 212) allow for individual pressurization of each of these cavities with gas when the respective cavities are enclosed after inserting the frame (10) and tray portion (15) into the corresponding housing of the device.

[0129] The cavity (224) for holding a flexible product container comprises a through-hole (27) shaped to accommodate at least a portion of the flexible container and at least a portion or all of the inlet port of said container, as well as a resealable outlet port. The cavity (225) for holding a flexible waste container comprises a through-hole (27) shaped to accommodate at least a portion of the flexible waste container and at least a portion or all of the inlet port of said container. The through-holes (27) provided in these cavities can be useful to accommodate the fill volume of the flexible product container and / or flexible waste container when the tray and its assembled components are in operation, for example, in the methods of preparing nanoparticles described herein.

[0130] As shown in Figures 3 and 4, a means (28) for holding a conduit is further provided, which in the illustrated example is a through-hole molded in the tray portion (15) of the frame (10) for holding or accommodating a conduit downstream of the outlet port of the static mixing device. The means (28) is adapted, for example, to hold a conduit immediately upstream of the inlet port of a flexible waste container and a flexible product container, respectively, and to hold one or more conduits or conduit sections immediately upstream of the outlet port of the static mixing device. As shown, the means (28) is adjacent to the cavities (224, 225) for holding the containers, specifically the through-holes (27) in each of the cavities, and also adjacent to the means (223) for holding the static mixing device, i.e., the recess (223), and a portion of the means (28) also abuts or crosses the means (18) for holding the valve.

[0131] Figures 3 and 4 also show securing means (26) for holding the containers. These may be, for example, but not limited to, protrusions or pins in the cavities (211, 212, 224, 225), and may be the same as, or may match or be complementary to, the means shown on the assembled frame and flexible container as shown in Figures 1 and 2. As shown, each cavity (211, 212, 224, 225) comprises at least four means (26) or points for securing the containers in the cavity.

[0132] 5 shows a perspective view, not drawn to scale, of another embodiment of a frame (50) according to the present disclosure having a horizontal operating orientation, in which the plane formed by the two longer dimensions of the frame is perpendicular to a vertical axis (100). The exemplary frame (50) is shown with a tray portion (55) forming substantially the entire frame, said frame (50) including: (a) a first flexible substrate container (51) and a second flexible substrate container (52), each having an outlet port (91) and a sealed inlet port (92), respectively; (b) a flexible waste container (53) having an inlet port (93); (c) a flexible product container (54) having an inlet port (94) and a resealable outlet port (95); and (d) a first and second inlet port and an outlet port (98). (e) conduits for fluidly connecting the outlet port (91) of the first substrate container (51) with the first inlet port of the static mixing device (16) and the outlet port (91) of the second substrate container (52) with the second inlet port of the static mixing device (16) (see the diagram shown in Figure 6 for further details); and (f) conduits (17c, 17d, respectively) for fluidly connecting the outlet ports of the static mixing device (16) with the inlet ports (94, 93) of the product container (54) and the waste container (53).

[0133] The illustrated frame (50) comprises cavities (551, 552, 524, 525; not all references are shown, but see also Figures 6 and 7) shaped to hold each of the containers. In the illustrated embodiment, the first cavity (511) is shaped to hold the first flexible substrate container (51), at least a portion of the sealed inlet port (92), the outlet port (91), a conduit connecting the outlet port (91) of the container (51) with the first inlet port of the static mixing device (16), and the inlet port of the static mixing device (16) itself. The second cavity (552) is shaped to hold the second flexible substrate container (52), the sealed inlet port (92), the outlet port, and at least a portion of the conduit connecting the outlet port of the second flexible container (52) with the second inlet port of the static mixing device (16) and the second inlet port of the static mixing device. The cavity (524) for holding the flexible product container (54) is also shaped to accommodate at least a portion of the inlet port (94) and at least a portion of the reclosable outlet port (95) of said container. The cavity (525) for holding the flexible waste container (53) is also shaped to accommodate at least a portion of the inlet port (93) of said container.

[0134] In this embodiment of the frame (50) according to the present disclosure, the first and second flexible substrate containers (51, 52) and the static mixing device (16) are held in the cavities (551, 552) of the tray portion (55) of the frame (50) in an arrangement such that, in their horizontal operating orientation, fluid flow from the static mixing device (16) through its outlet port toward the waste or product containers (54, 53) occurs at least partially in an anti-gravity direction. Furthermore, the means for holding the first and second cavities (551) and the static mixing device (16) are positioned on the tray portion (55) such that the outlet ports (91) of both flexible substrate containers have a lower position relative to the vertical axis (100) relative to the positions of the inlet ports (93, 94) of the flexible waste container (53) and the flexible product container (54), respectively.

[0135] Figure 6 is a top view (i.e., on the vertical axis (100)) of the same frame (50) as described in Figure 5. This view is also not drawn to scale, but shows the circumferential gaskets (19) characteristic of the first and second cavities (see also Figure 7). Each of these gaskets (19) is conformally shaped to its respective cavity and is used to form a seal on those cavities when an appropriate counterpiece device is applied thereon.

[0136] As shown and referenced in this figure, the containers (51, 52, 53, 54) are each fixed in their respective cavities by a plurality of means (26) for fixing the containers. As illustrated in this depiction, up to four means (26) for fixing are provided, with fixing points located in the peripheral zone of the containers, on or near at least two corners of the sealing edge of the containers.

[0137] As further shown in this depiction, the first flexible substrate container (51) and the second flexible substrate container (52) are held by the arrangement of the first and second cavities, respectively, on the tray portion (55) of the frame (50), such that the outlet port (91) of the first flexible substrate container (51) and the outlet port (91) of the second flexible substrate container (52) are coaxially opposed (see also FIG. 7). In particular, the outlet port (91) of the first flexible substrate container (51) and the outlet port (91) of the second flexible substrate container (52) are coaxially opposed at an angle of approximately 45° relative to the central longitudinal axis (not shown) of the frame (50). Conduits 17a, 17b are also substantially coaxially opposed for fluidly connecting the outlet port (91) of the first substrate container (51) with the first inlet port (96) of the static mixing device (16) and the outlet port (91) of the second substrate container (52) with the second inlet port (97) of the static mixing device (16), respectively.

[0138] Similarly, the flexible waste container (53) and the flexible product container (54) are each held by the arrangement of their corresponding cavities on the tray portion (55) of the frame (50), so that the inlet ports (93, 94) of each of these containers are also coaxially opposed (see also FIG. 7). The arrangement provided by the frame (50) also provides that at least a portion or a majority (i.e., at least 50% of the longitudinal length) of the conduits (17c, 17d) for fluidly connecting the outlet port (98) of the static mixing device (16) to the flexible product container (54) and the flexible waste container (53), respectively, are also coaxially opposed.

[0139] Figure 7 shows the same top view of the frame (50) and its tray portion (55) described in Figure 6, without the assembled components of the first and second flexible substrate containers, flexible waste and flexible product containers, static mixing device, and their associated conduits. Shown are a cavity (551) for holding a first flexible substrate container, a cavity (552) for holding a second flexible substrate container, a cavity (525) for holding a flexible waste container, and a cavity (524) for holding a flexible product container. Also shown is a means (523) for holding a static mixing device, which is a recess in the tray portion (55) of the frame (50). The recess is also adjacent to the first cavity (551) and the second cavity (552) and is adapted to hold the static mixing device so that fluid flow immediately exiting the outlet port of the static mixing device is in the anti-gravity direction. Each of the first and second cavities (551, 552) is individually provided with a circumferential gasket (19) in said corresponding cavity, a portion of each gasket of each cavity being provided so as to be sealable against a portion of the static mixing device and its respective inlet port. Also shown are means (28) for retaining conduits, specifically conduits for fluidly connecting the outlet ports of the static mixing device to the flexible product container (54) and flexible waste container (53), respectively, which are also provided in the form of recesses in the tray (55) portion of the frame (50).

[0140] 8 shows a perspective view, not drawn to scale, of another embodiment of a frame (80) according to the present disclosure having a horizontal operating orientation, in which the plane formed by the two longer dimensions of the frame (80) is perpendicular to a vertical axis (100). The exemplary frame (80) is shown comprising a tray portion (85) forming substantially the entirety of the frame (80), said frame comprising: (a) a first flexible substrate container (81) and a second flexible substrate container (82), each having an outlet port (91) and a sealed inlet port (92), respectively; (b) a flexible waste container (83) having an inlet port (93); (c) a flexible product container (84) having an inlet port (94) and a resealable outlet port (95); and (d) a first inlet port (96) and a second inlet port (97), and an outlet port (98). ), and (e) conduits (17a, 17b, respectively) for fluidly connecting the outlet port (91) of the first substrate container (81) with the first inlet port (96) of the static mixing device (16) and the outlet port (91) of the second substrate container (82) with the second inlet port (97) of the static mixing device (16), and conduits (17c, 17d, respectively) for fluidly connecting the outlet port (98) of the static mixing device (16) with the inlet ports (94, 93) of the flexible product container (84) and the flexible waste container (83). The frame also comprises means (18) for holding valves.

[0141] The illustrated frame includes cavities (881, 882, 824, 825) molded to hold each of the containers. In the illustrated embodiment, the first cavity (881) is molded to hold a first flexible substrate container (81), its sealed inlet port (92), outlet port (91), a conduit (17a) connecting the outlet port of the first container with the first inlet port (96) of the static mixing device (16), and the inlet port (96) of the static mixing device (16). The second cavity (882) is molded to hold a second flexible substrate container (82), its sealed inlet port (92), outlet port (91), a conduit (17b) connecting the outlet port of the second flexible container (82) with the second inlet port (97) of the static mixing device (16), and the second inlet port (97) of the static mixing device (16). The first cavity (881) and second cavity (882) each also include a circumferential gasket (19) that is conformally shaped to the respective cavity and that is used to form a seal thereon when an appropriate counterpiece is applied thereon. The cavity (824) for holding a flexible product container (84) is also shaped to accommodate the container (84), at least a portion of the container's inlet port (94), and at least a portion of the container's resealable outlet port (95). The cavity (825) for holding a flexible waste container (83) is also shaped to accommodate at least a portion of the container's inlet port (93).

[0142] The containers (81, 82, 83, 84) are each fixed in their respective cavities by a plurality of means (26) for fixing the containers. As illustrated in the present depiction, the means (26) for fixing are provided at two locations on the containers in the peripheral zone (not all of which are shown, see also Figures 9, 10 and 11), or at the sealing edge of the containers.

[0143] In this illustrated embodiment of a frame (80) according to the present disclosure, the first and second flexible substrate containers (81, 82) and the static mixing device (16) are held in a cavity in the tray portion (85) of the frame (80) in an arrangement such that, in their horizontal operating orientation, fluid flow from the static mixing device (16) through its outlet port (98) toward the waste or product containers (83, 84) occurs at least partially in an anti-gravity direction.

[0144] Figure 9 shows a top view (i.e., on the vertical axis (100)) of the same frame (80) as described in Figure 8. This view, again not drawn to scale, shows circumferential gaskets (19) contained in first cavity (881) and second cavity (882), respectively. Each of these gaskets (19) is adapted to its respective cavity and is used to form a seal on those cavities when an appropriate counterpiece device is applied thereon.

[0145] As shown and referenced in this figure, the containers (81, 82, 83, 84) are each secured in their respective cavities by a plurality of means for securing the containers (26). As illustrated in this depiction, two means for securing (26) are provided per cavity, with the securing points located in the peripheral zone of the container, proximate to the substrate container outlet (91) or the product or waste container inlet (94, 93).

[0146] As further shown in this depiction, the first flexible substrate container (81) and the second flexible substrate container (82) are held by the arrangement of the first cavity (881) and the second cavity (882), respectively, on the tray portion (85) of the frame (80), such that the outlet port (91) of the first flexible substrate container (81) and the outlet port (91) of the second flexible substrate container (82) are coaxially opposed. Conduits for fluidly connecting the outlet port of the first substrate container (81) with the first inlet port (96) of the static mixing device (16) and the outlet port of the second substrate container (82) with the second inlet port (97) of the static mixing device (16), respectively, are also substantially coaxially opposed.

[0147] Figure 10, not drawn to scale, shows the same perspective view of the frame (80) of Figure 8 in a horizontal operating position perpendicular to the vertical axis (100) but with the plane formed by the two longest dimensions of the frame, without any of the first and second flexible substrate containers, flexible waste container and flexible product container, static mixing device, and associated conduits.

[0148] Figure 11, also not drawn to scale, shows a top view of the same frame (80) as Figure 9, without the assembled first and second flexible substrate containers, flexible waste container and flexible product container, static mixing device, and any associated conduits.

[0149] Both Figures 10 and 11 show a) a first cavity (881) shaped to hold a first flexible substrate container and at least a portion of its outlet port and sealed inlet port, and conduits for fluidly connecting the outlet port of said container to a first inlet port of the static mixer and a first inlet port of the static mixing device; and b) a second flexible substrate container and at least a portion of its outlet port and sealed inlet port, and conduits for fluidly connecting the outlet port of said container to a second inlet port of the static mixing device and a second inlet port of the static mixing device. An exemplary frame (80) is shown comprising a tray portion (85) comprising: a second cavity (882) for holding a tube; and c) means (823) for holding a static mixing device, the means being a recess in the tray portion (85) adapted to accommodate at least a portion or a partial profile of the static mixing device, and adapted, in conjunction with the operational orientation of the frame (80), to hold said device in an arrangement such that fluid flow immediately exiting the outlet port of the mixing device is at least partially in an anti-gravity direction. In the unassembled state of the frame (80), i.e., without the substrate container and mixing device, said recess (823) is further in fluid communication with the first cavity (881) and with the second cavity (882).

[0150] As shown in Figures 10 and 11, the first and second cavities are each provided with a circumferential gasket (19) in the corresponding cavity, with a portion of each gasket in each cavity being adapted to be sealable against a portion of the static mixing device and its respective inlet port.

[0151] As shown in Figures 10 and 11, means for retaining or partially retaining conduits (28) are further provided. The means for retaining conduits (28) for fluidly connecting the first and second containers to the inlet ports of the static mixing device are provided as recesses in the tray portion (85). Means (28) adapted to at least partially retain conduits immediately upstream of the inlet ports of the flexible waste container and the flexible product container, respectively, are also provided in the form of recesses in the tray portion (85). The means for retaining valves (18) may also further comprise means for retaining conduits downstream of the outlet port of the static mixing device.

[0152] Figures 10 and 11 also show fixing means (26) for holding the container. As shown in Figure 10, the cavities (881, 882, 824, and 825) are provided with arrow-head or mushroom-head protrusions, or barbed pins or plugs, which are adapted to receive a flexible container having through holes positioned to match said protrusions (for reference, see corresponding locations of fixing means in Figures 8 and 9, which show a frame and assembled with a flexible container).

[0153] The cavity (881) for holding the first flexible substrate container and the cavity (882) for holding the second flexible secondary container further comprise gas pressure distribution means (29). When the cavities are surrounded by a counterpiece device and the means for forcing the liquid substrate from the first and second flexible substrate containers to flow into the static mixing device are provided in the form of pressurized gas (e.g., pressurized air), said means may be useful for uniformly distributing the air flow across the cavities and over the flexible containers. The gas pressure distribution means (29) may also be provided as a series of vents adapted to the shape of the cavities in which the flexible substrate containers may be placed. In an alternative embodiment, the cavities (881, 882) comprise a plurality of laterally arranged through-holes, for example in the form of slits or vents, which allow the pressurized air to diffuse into the cavities.

[0154] 12 shows a front view, not drawn to scale, of another example of a frame (120) according to the present disclosure. The frame (120) is shown in its operational orientation, which is vertical and parallel to the vertical axis (100). The frame (120) comprises a tray portion (125) that simultaneously holds a first flexible substrate container (121) and a second flexible substrate container (122), each substrate container having an outlet port (91), and a static mixing device (16) that includes a first inlet port (96), a second inlet port (97), and an outlet port (98). The illustrated frame (120) further simultaneously holds a flexible waste container (123) including an inlet port (93), a flexible product container (124) including an inlet port (94), conduits (17, 17c, 17d) for fluidly connecting the outlet port (98) of the static mixing device (16) with the inlet ports (94, 93) of the product container (124) and the waste container (123), and a valve (20) in which the conduits (17, 17c, 17d) are disposed for directing fluid flow from the outlet port (98) of the static mixing device (16) to the inlet port (93) of the waste container (123) or the inlet port (94) of the product container (124). A circumferential gasket (19) is provided for each of the first and second cavities, which can be used to form a seal on these cavities when an appropriate counterpiece is applied thereon.

[0155] As shown, the first and second flexible substrate containers (121, 122) and the static mixing device (16) are held in the cavity of the tray portion (125) of the frame (120) in an arrangement such that, in the illustrated operational orientation, fluid flow from the outlet (98) of the static mixing device (16) to the waste or product containers (123, 124) occurs at least partially in an anti-gravity direction. The frame (120) may, in some embodiments, be arranged so that it is reversibly securable to an apparatus as defined herein, such as an apparatus for filling substrate containers. The frame (120) in its operational orientation is also arranged so that the outlet ports (91) of both flexible substrate containers have a lower position relative to the positions of the respective inlet ports (93, 94) of the flexible waste container (53) and flexible product container (54) with respect to the vertical axis (100).

[0156] 13 shows an example of a flexible container (1) for use with a frame or tray portion of a frame, or apparatus, or provided as part of a kit, according to any one of the embodiments or combinations of embodiments described herein. The flexible container (1) may be useful as a flexible substrate container, flexible waste container, or flexible product container, and is adapted to be reversibly retained within a corresponding cavity in the frame. The flexible container includes at least one inlet or outlet port (2) and, in this embodiment, a sealable port (3), which may be sealed depending on the function of the container or its usage, such as the method of use of the frame or apparatus described herein.

[0157] In the illustrated embodiment, the flexible container comprises an interior space (4) for holding a fluid material surrounded by flexible walls formed by front and rear walls made of a polymeric material, the flexible front and rear walls being connected to one another to form a sealing edge (7) that substantially encloses the interior space (4), the edge (7) comprising four corner regions, with a through-hole (27) provided at or near a first corner region and a through-hole (27) provided at or near a second corner region of the sealing edge (7), the second corner region being adjacent to the first corner region. In the currently illustrated example, the flexible container comprises a through-hole (27) in each corner region, the through-holes positioned to coincide with the positions of corresponding fastening or retaining means provided in corresponding cavities of a frame or tray portion of a frame according to the present disclosure.

[0158] Figure 14 is a perspective side view of the flexible container (1) of Figure 13 showing the end of the container with ports (2, 3) in fluid communication with an interior space (4) for holding a fluid material, said space being formed by a flexible front wall (5) and a flexible rear wall (6), each wall being connected to form a continuous sealing edge (7).

[0159] Figure 15, not drawn to scale, shows a front or user-facing side view of another example of a frame (150) according to the present disclosure. The frame (150) is shown in its operational orientation, vertical and parallel to the vertical axis (100). The figure also shows an example of a kit according to the present disclosure, where the frame (150) is assembled to hold flexible containers (11, 12, 14), a conduit (17), and a static mixing device (16). The frame (150) includes a first sealable area (231) and a second sealable area (232) for holding first and second flexible substrate containers (11, 12). The sealable areas (231, 232) are defined by a circumferential gasket (19). The sealable areas (231, 232) are positioned adjacent to one another. Although not drawn to scale, it can be seen that the first sealable area (231) is larger than the second sealable area (232), the size difference being reflected in that the sealable areas (231, 232) have the same height, but the width of the first sealable area (231) is larger than the width of the second sealable area (232). Within the sealable areas (231, 232) of the frame (150), the first and second flexible substrate containers (11, 12) are held in their designated positions by means (226). The means (226) are connectable with corresponding through-holes (see 228) in the peripheral zone or sealing edge (227) of each flexible container. Each substrate container (11, 12) has three ports, including an outlet port (91) and a sealing inlet port (92). The outlet port (91) is fluidly connected to the static mixing device (16) via a conduit (17), and the static mixing device is held in place by a means for holding it (223). Furthermore, the outlet port (91) is located at the top of the substrate containers (11, 12) so that the fluid substrate or liquid (not shown) held therein exits the substrate containers (11, 12) in an anti-gravity direction to flow to the static mixing device (16). Also shown is a flexible product container (14) positioned on and towards the front of the frame (150) and held by three fixing means (226) whose positions correspond to three through-holes (228) provided in the peripheral zone of the product container (14).Note that the flexible waste container is secured to the rear surface of the frame and therefore cannot be seen (see FIG. 16). The product container (14) has three ports located on its bottom surface (its operational orientation), including an inlet port (94) for receiving a third fluid from the static mixing device (16) via a conduit (17) and two outlet ports (95). One of the outlet ports (95) is fluidly connected via flexible tubing to a sampling tube (241), the downstream end of which is fluidly connected to a sterile filter (242). A pinch valve (246) and a sterile disconnector (247) are located upstream of the sampling tube (241) to facilitate the withdrawal of a product sample. The other outlet port (95) is fluidly connected via flexible tubing to the sterile connector (245). Another sterile filter (249) is also shown in fluid communication with the product container's inlet port via a Y-piece (248). This arrangement may be used to add diluent to product container (14) before, during, or after the mixing process to dilute or alter the composition of the third fluid received from static mixing device (16). Another advantageous use is to inject enough diluent or sterile air necessary to push any remaining (third) fluid present in conduit (17) between Y-piece (248) and inlet port (94) into product container (14) to maximize the yield of the mixing process. Frame (150) further comprises four through-holes (251) in the central region that allow frame (150) or kit to be secured to an apparatus for operation.

[0160] FIG. 16 shows a front view of the rear of the same frame (150) or kit shown in FIG. 15, also shown in an operational orientation that is vertical and parallel to the vertical axis (100). Here, a flexible waste container (13) can be seen secured to the rear of the frame. In the operational orientation, the bottom of the waste container (13) presents three ports, including an inlet port (93) fluidly connected via a conduit (17) to a valve (20), which is a one-way stopcock valve. The valve (20) is located downstream of a T-piece (21) through which a third fluid is received from a static mixing device. The T-piece (21), whose inlet is fluidly connected to the static mixing device, has a second outlet downstream of another valve (20), also a one-way stopcock valve, located and fluidly connected to a Y-piece (248), which is fluidly connected to a sterile filter (249) and a flexible product container secured to the front of the frame (150) (see FIG. 15). Also, a circumferential gasket (19) is arranged on the rear surface of the frame (150) surrounding the through-hole (27) in the same manner as on the front surface (as shown in FIG. 15).

[0161] FIG. 17 shows a front elevational view of the frame (150) shown in FIG. 15 in an operational orientation that is vertical (parallel to the vertical axis (100)), except that it is absent any other components that define flow paths for any of the first, second, or third liquids, such as flexible containers, conduits, static mixing devices, or T- or Y-pieces, valves, or filters. In other words, it is a depiction of the exemplary frame (150) before assembly into a kit according to some embodiments disclosed herein. Shown are protruding through-holes (27) in the first and second sealable areas (231, 232) through which pressurized gas can be supplied in an operational state to apply pressure to the exterior surfaces of the first and second flexible substrate bags. Also shown are recesses (243, 244) for holding sampling tubes and their associated sterile filters and sterile connectors, respectively. The frame (150) further includes means (233) for securing the frame to an external holding means, such as a hook, to assist a user in accurately positioning the frame (150) on any corresponding holding means that may be provided by, for example, an apparatus for filling a flexible substrate container when secured to the frame (150) or an apparatus for operating the fully assembled frame (150) or kit. A grip or handle (234) is provided to allow a user to easily hold and position the frame (150). Various small openings or through-holes (235) are shown for holding conduits or allowing conduits to connect the front and rear sides of the frame (150). The sealable areas (231, 232) are provided with means (253) for securing or holding at least one conduit to the frame, or more specifically, for holding a conduit within the sealable area. The one or more conduits may be associated with, for example, a container inlet port (e.g., a crimped tube end), and the means (253) may, in some embodiments, be a hook.

[0162] Figure 18 is a front view of the rear face of the frame (150) shown in Figure 17 in an operational orientation parallel to the vertical axis (100). As shown, fixing or fastening means (26) for flexible waste containers are provided on the rear face of the frame (150). Additionally, two means (252) for fixing or retaining the valves (20) are provided, as described in Figure 16. Means for fixing a conduit (253) to the frame are also shown.

[0163] Figure 19 shows a perspective view of the front of the frame (150) shown in Figure 15. Here, it can be seen how, in this embodiment, a circumferential gasket (19) is provided on a circumferential rim (22) extending from the frame (150) that surrounds the first and second sealable areas (231, 232) of the frame (150).

[0164] Figure 20 shows a perspective view of the rear of the frame (150) shown in Figure 19. In this embodiment, the rear of the frame (150) is also shown to have a circumferential gasket (19) provided on the circumferential rim (22).

[0165] Figure 21 shows a detailed front view of a portion of the rear face of the frame (150) shown in Figure 16. The two outlets of the T-piece (21), located downstream of and in an upper position relative to the static mixing device (16), are connected to valves (20), such as one-way stopcock valves, that can open and close the flow paths to the product and waste containers (13). The valves (20) exhibit means (23) for mechanically operating the valves (20) that are oriented in a direction corresponding to the rear face of the frame (150) and adapted to operate from that direction. Such an arrangement allows the valves (20) to be operated, for example, by an automatic operator device located behind the frame (150) when the frame (150) or kit is inserted into an apparatus adapted to operate the frame (150) or kit, i.e., when viewed from the front.

[0166] Figure 22 is a perspective view of an exemplary static mixing device (162) that can be used in combination with, for example, the frame (150) or kit shown in Figures 15 and 16. The static mixing device (162) may represent a jet impingement reactor, according to some preferred embodiments. The illustrated mixing device (162) comprises a main housing (30) having an outlet port (31) and first and second inlet ports (32, 33). In this case, the static mixing device (162) is shown in its operational orientation with the outlet port (31) facing upward so that the third fluid exiting the outlet port (31) flows in an anti-gravity direction. First and second inlet connecting pieces (34, 35) fit into the first and second inlet ports (32, 33), respectively. Barbed connectors (36) are provided at the upstream ends of the inlet ports (32, 33) and the downstream end of the outlet port (31).

[0167] Figure 23 is a perspective view of the exemplary static mixing device (162) shown in Figure 22. Here, the main housing (30) having the outlet port (31) and first and second inlet ports (32, 33) is shown without the first and second inlet connecting pieces.

[0168] FIG. 24 is a perspective view of the front or user-facing side of another example of a frame (240) according to the present disclosure, not drawn to scale. Any other components described herein that define flow paths for any of the first, second, or third liquids, such as flexible containers, conduits, static mixing devices, T-pieces or Y-pieces, valves, or filters, are not present in this view. In other words, it is a depiction of the exemplary frame (240) before it is assembled into a kit according to some embodiments disclosed herein. The frame (240) is shown in its operational orientation, which is vertical and parallel to the vertical axis (100). The frame (240) includes a first sealable area (231) and a second sealable area (232) for holding first and second flexible substrate containers (not shown). The sealable areas (231, 232) are defined by a circumferential gasket (19) provided by a circumferential rim (22) extending from the frame. The sealable areas (231, 232) are positioned adjacent to one another. While not drawn to scale, it can be seen that the first sealable area (231) is larger than the second sealable area (232), with the size difference reflected in that the sealable areas (231, 232) have the same height, but the width of the first sealable area (231) is larger than the width of the second sealable area (232). In this embodiment, the shape of the first sealable area (231), like the second sealable area (232), is symmetrical about a vertical axis. Also shown are through-holes (27) in the first and second sealable areas (231, 232) through which pressurized gas can be supplied in an operative state to apply pressure to the outer surfaces of the first and second flexible substrate bags. Also shown within the first and second sealable areas are means (226) for securing flexible substrate containers.

[0169] Additionally, recesses (243, 244) are shown for holding sampling tubes, their associated sterile filters, and sterile connectors, respectively. Frame (240) further includes means (233) for securing the frame to an external holding means, such as hooks or recesses, to assist the user in accurately positioning frame (240) on any corresponding holding means that may be provided by, for example, an apparatus for filling flexible substrate containers when secured to frame (240), or an apparatus for operating the fully assembled frame (240) or kit. A grip or handle (234) is further provided to allow the user to easily hold and position frame (240). Means (226) for securing product containers to frame (240) are also provided, for example, in the upper region of the frame in this illustrated embodiment. Various small openings or through-holes (235) are also shown for holding conduits or allowing conduits to connect the front and rear of frame (240). Within the sealable area (231, 232) of the frame, means (253) are provided for securing or retaining conduits to the frame and within said sealable area. In particular, one or more conduits associated with a container inlet port (e.g., a crimped tube end) may be retained and accommodated within said sealable area of ​​the frame by such means, for example, hooks. The frame (240) further comprises four through-holes (251) in the central area that allow for fixing the frame (240) or the kit to a device for operating it.

[0170] Figure 25 shows a perspective view of the rear face of the frame (240) shown in Figure 24 in a vertical operating orientation parallel to the vertical axis (100). As shown, fixing or fastening means (26) for flexible waste containers are provided on the rear face of the frame (240). The sealable areas (231, 232) for the rear face of the frame (240) are also defined by circumferential gaskets (19) provided by circumferential rims (22) extending from the frame. The circumferential gaskets (19) provided on the rear face of the frame (240) may advantageously be of the same shape and dimensions as the gaskets (19) used on the rims of the respective sealable areas on the front face of the frame (see Figure 24). Two means (252) for securing valves, such as those shown in Figures 16 and 21, to the frame (240) are also shown. Means (253) for securing conduits to the frame (240) are also shown. [Explanation of symbols]

[0171] 1 flexible container 2 inlet / outlet ports 3 Sealable Ports 4. Internal volume 5 Flexible front wall 6 Flexible back wall 7 Sealing Edge 10, 50, 80, 120, 150, 240 frames 11,51,81,121 First flexible substrate container 12,52,82,122 Second flexible substrate container 13,53,83,123 Flexible waste containers 14,54,84,124 Flexible product container 15,55,85,125 Tray part 16,162 static mixing devices 17,17a,17b,17c,17d conduit 18 Means for retaining valve 19 Circumferential gasket 20 valves 21 T-piece 22 Circumferential Rim 23 Means for operating valves 26 Fixing or fastening means 27 For example, through holes in sealable areas or cavities 28 Means for retaining conduit 29 Gas pressure distribution means 30 Main housing of static mixing device 31 Static Mixing Device Outlet Port 32 First inlet port of static mixing device 33 Second inlet port of static mixing device 34 First inlet connecting piece 35 Second inlet connecting piece 36 Barbed Connector 91 outlet port of first or second flexible substrate container 92 sealed inlet port of first or second flexible substrate container 93 Flexible waste container inlet port 94 Flexible product container inlet port 95 Resealable outlet port of flexible product container 96 First inlet port of static mixing device 97 Second inlet port of static mixing device 98 Static Mixing Device Outlet Port 100 vertical axis 211,551,881 A first cavity for holding a first flexible substrate container 212,552,882 A second cavity for holding a second flexible substrate container 223,523,823 Means for maintaining static mixing devices 224,524,824 Cavity for flexible product container 225,525,825 Cavities for flexible waste containers 226 Means for securing flexible substrates, product, or waste containers 227 Sealing edges of flexible substrates, product, or waste containers 228 Perforations in the peripheral zone of flexible containers 231 a first sealable area for holding a first flexible substrate container 232 a second sealable area for holding a second flexible substrate container; 233 Means for fastening the frame to the external retaining means 234 Handle 235 Through hole 241 Sampling tube 242 Sterile Filter 243 Recesses for holding sampling tubes and sterile filters 244 Recess to hold sterile connector 245 Sterile Connector 246 Pinch Valve 247 Sterile Disconnector 248 Y-Piece 249 Sterile Filter 251 Through holes for matching hooks that hold the frame in place 252 Means for fixing valves 253 Means for retaining or fastening conduit to a frame

[0172] The following numbered item list are further embodiments included in the present disclosure:

[0173] 1. (a) first and second flexible substrate containers, each having an exit port; (b) a flexible waste container with an inlet port; (c) a flexible product container with an inlet port; (d) a static mixing device having at least first and second inlet ports and an outlet port; (e) a frame for simultaneously holding conduits for fluidly connecting the outlet port of the first substrate container with the first inlet port of the static mixing device, the outlet port of the second substrate container with the second inlet port of the static mixing device, and the outlet port of the static mixing device with the inlet ports of the waste container and the product container; The frame is a first cavity for holding a first flexible substrate container; a second cavity for holding a second flexible substrate container; - a tray portion comprising means for holding a static mixing device; The frame has an operational orientation, and the means for holding the first and second cavities and the static mixing device are arranged so that fluid flow from the first and second substrate containers to the static mixing device in the operational orientation, and / or fluid flow from the static mixing device to the waste container or product container, occurs at least partially in an anti-gravity direction.

[0174] 2. The frame of item 1, wherein the tray portion includes a further cavity for a flexible product container and a further cavity for holding a flexible waste container.

[0175] 3. The frame according to item 1 or 2, wherein each of the first and second cavities comprises one or more means for securing a respective flexible substrate container, said means optionally being shaped as protrusions or pins for receiving the flexible substrate container having through holes positioned to match the pins or protrusions, and the pins are optionally barbed pins or snap-lock pins.

[0176] 4. The frame of any one of items 1 to 3, further comprising an identification tag such as an RFID tag.

[0177] 5. A frame described in any one of items 1 to 4, wherein the means for holding the static mixing device is adapted to hold the static mixing device only when said static mixing device has a desired orientation, and when the static mixing device has its desired orientation and the frame is in its operating orientation, flow of liquid from the static mixing device through its outlet port optionally occurs in an anti-gravity direction.

[0178] 6. A frame according to any one of items 1 to 5, wherein the means for holding the first and second cavities and the static mixing device are arranged such that, in the operational orientation of the frame, the outlet ports of the flexible substrate container have inferior positions relative to the positions of the respective inlet ports of the static mixing device.

[0179] 7. The tray part is a first through-hole disposed in the first cavity and a second through-hole disposed in the second cavity; and / or - a circumferential gasket for each of the first and second cavities The frame according to any one of items 1 to 6, comprising:

[0180] 8. A frame according to any one of items 1 to 7, wherein the first cavity is shaped to at least partially accommodate a conduit for fluidly connecting an outlet port of a first substrate container with a first inlet port of a static mixing device, or the second cavity is shaped to at least partially accommodate a conduit for fluidly connecting an outlet port of a second substrate container with a second inlet port of a static mixing device.

[0181] 9. The frame is adapted to be insertable in its operational orientation into an apparatus for aseptically mixing two fluids, the apparatus comprising: a counterpiece for sealingly covering the first and second cavities; - means for applying pressure to the first and second flexible substrate containers when inserted into the first and second cavities; 9. The frame of any one of items 1 to 8, wherein the frame optionally holds first and second substrate containers, a flexible waste container, a flexible product container, a static mixing device, and conduits for fluidly connecting an outlet port of the first substrate container with a first inlet port of the static mixing device, for fluidly connecting an outlet port of the second substrate container with a second inlet port of the static mixing device, and for fluidly connecting an outlet port of the static mixing device with inlet ports of the waste container and the product container.

[0182] 10. A kit comprising a frame according to any one of items 1 to 9, (a) first and second flexible substrate containers; (b) a flexible waste container; (c) a flexible product container; (d) a static mixing device; (e) a conduit for fluidly connecting the outlet port of the first substrate container with the first inlet port of the static mixing device, the outlet port of the second substrate container with the second inlet port of the static mixing device, and the outlet port of the static mixing device with the inlet ports of the flexible waste container and the flexible product container, and optionally for filling each of the first and second flexible substrate containers with liquid substrate.

[0183] 11. The static mixing device comprises a T-piece mixer, a Y-piece mixer, a vortex mixer, a baffle-based static mixer, a microfluidic mixing device, a multi-inlet vortex mixer (MIVM), or a jet impingement reactor, the jet impingement reactor optionally comprising: a reaction chamber defined by an inner surface of a reaction chamber wall, the reaction chamber having a substantially spheroidal overall shape, said chamber comprising: (a) first and second fluid inlets, the first and second fluid inlets being arranged at opposing positions on a first central axis (x) of the reaction chamber so as to face each other, and each of the first and second fluid inlets comprising a nozzle; (b) a fluid outlet disposed at a third location, the third location being on a second central axis of the chamber, the second central axis being perpendicular to the first central axis; Item 11. The kit of item 10, wherein a distance (d) between the nozzle of the first fluid inlet and the nozzle of the second fluid inlet is equal to or smaller than a diameter of the reaction chamber along the first central axis.

[0184] 12. A flexible container useful as a flexible substrate container, flexible waste container, or flexible product container according to any one of items 1 to 11, comprising: an interior space for holding a fluid material surrounded by a flexible front wall and a flexible rear wall, each wall being made of a polymeric material; and at least one inlet or outlet port for allowing fluid communication with the interior space, wherein the flexible front wall and the flexible rear wall are connected to each other to form a sealing edge that substantially encloses the interior space, the edge including four corner regions such that when empty the interior space has an overall square or rectangular shape, wherein a first through-hole is provided at or near the first corner region of the sealing edge, and a second through-hole is provided at or near the second corner region of the sealing edge, the second corner region being adjacent to the first corner region.

[0185] 13. An apparatus comprising: a housing adapted to receive the frame described in item 9 in its operational orientation, the frame having the first and second flexible substrate containers empty; and means for aseptically filling the first and second substrates into the first and second flexible substrate containers.

[0186] 14. An apparatus comprising: a housing adapted to receive the frame described in item 9 in its operative orientation, wherein the first and second flexible substrate containers contain the first and second substrates; and means for forcing the substrates to flow from the first and second flexible substrate containers into a static mixing device so as to mix and form a liquid product, said means optionally comprising pressurized gas.

[0187] 15. A method for mixing two fluid substrates, comprising the use of a frame according to any one of items 1 to 9, a kit according to item 10 or 11, a flexible container according to item 12, or an apparatus according to item 13 or 14.

Claims

1. (a) first and second flexible substrate containers, each having an exit port; (b) a flexible waste container with an inlet port; (c) a flexible product container with an inlet port; (d) a static mixing device comprising at least first and second inlet ports and an outlet port; (e) a frame adapted to simultaneously hold conduits for fluidly connecting the outlet port of the first substrate container with the first inlet port of the static mixing device, the outlet port of the second substrate container with the second inlet port of the static mixing device, and the outlet port of the static mixing device with the waste container and the inlet ports of the product container; The frame is a first sealable area for holding said first flexible substrate container; a second sealable area for holding said second flexible substrate container; - means for holding said static mixing device, The frame has an operational orientation, and the means for holding the first and second sealable areas and the static mixing device are arranged so that in the operational orientation, fluid flow from the first and second flexible substrate containers to the static mixing device and / or fluid flow from the static mixing device to the flexible waste container or flexible product container occurs at least partially in an anti-gravity direction.

2. The frame of claim 1 , wherein the first and / or second sealable areas are molded as cavities.

3. The frame of claim 1 or 2, wherein the first and / or second sealable areas are defined by a circumferential rim extending from the frame.

4. The frame of claim 3 , wherein the circumferential rim is arranged to sealingly receive a counterpiece having a cavity for receiving each of the flexible substrate containers.

5. 3. The frame of claim 2, wherein the frame comprises a tray portion, the tray portion comprising a first cavity for holding the first flexible substrate container, a second cavity for holding the second flexible substrate container, and optionally means for holding the static mixing device.

6. A frame according to any preceding claim, wherein the conduit is a non-microfluidic conduit.

7. A frame according to any preceding claim, wherein the conduit is flexible and reversibly connectable with the port to which it is adapted for fluid connection.

8. The frame of any one of claims 1 to 7, wherein the static mixer is adapted to mix fluids at a total flow rate of about 10 mL to about 1,000 mL / min.

9. 9. The frame of any one of claims 1 to 8, wherein each of the first and second sealable areas is shaped and dimensioned to hold a flexible substrate container having an internal volume in the range of 50 mL to 1,500 mL.

10. A frame according to any one of claims 1 to 9, wherein the first flexible substrate container has an internal volume that is 1.5 to 4 times larger than the internal volume of the second flexible substrate container.

11. The frame of any one of claims 1 to 10, wherein the first and second sealable areas have substantially similar or identical heights and substantially different widths.

12. A frame according to any one of the preceding claims, wherein the frame comprises means for securing the flexible product container and / or means for securing the flexible waste container.

13. 13. The frame of claim 12, wherein the frame has a front side facing the user in its operational orientation and a rear side opposite the front side, and the means for securing the flexible product container is located on the front side of the frame, and the means for securing the flexible waste container is located on the front side or the rear side of the frame.

14. A frame according to any preceding claim adapted for a vertical operating orientation.

15. A frame according to any preceding claim, wherein each of the first and second sealable areas comprises one or more means for securing the respective flexible substrate container.

16. The frame of claim 15 , wherein the one or more means for securing the flexible substrate container are disposed on the front surface of the frame.

17. 17. A frame according to claim 15 or 16, wherein the means is optionally shaped as one or more hooks, protrusions or pins arranged to receive a flexible substrate container having through holes positioned to align with the hooks, pins or protrusions.

18. 18. The frame of claim 17, wherein the pin is optionally a barbed pin or a snap lock pin.

19. 19. The frame of claim 1, wherein the first sealable area and the second sealable area are located adjacent to each other, and the minimum distance between the first sealable area and the second sealable area is less than 10% of the width of the first sealable area.

20. The frame of any one of claims 1 to 19, wherein the flexible product container has an internal volume in the range of 100 mL to 2,000 mL.

21. A frame according to any preceding claim, further comprising an identification tag, such as an RFID tag.

22. 22. A frame according to any one of claims 1 to 21, wherein the means for holding the static mixing device is adapted to hold the static mixing device only when the static mixing device has a desired orientation.

23. 23. A frame according to any one of claims 1 to 22, wherein the flow of liquid from the static mixing device through its outlet port occurs in an anti-gravity direction when the static mixing device has its desired orientation and the frame is in its operating orientation.

24. A frame according to any one of claims 1 to 23, wherein the first and second sealable areas and the means for holding the static mixing device are arranged such that, in the operating orientation of the frame, the outlet port of the flexible substrate container has a lower position relative to the position of the respective inlet port of the static mixing device.

25. A frame according to any one of claims 1 to 24, wherein the frame or the tray portion thereof comprises a first through hole disposed in the first sealable area and a second through hole disposed in the second sealable area.

26. A frame according to any preceding claim, wherein the frame or the tray portion thereof comprises a circumferential gasket for each of the first and second sealable areas.

27. 27. A frame according to any one of claims 1 to 26, wherein the first sealable area is shaped to at least partially accommodate the conduit for fluidly connecting the outlet port of the first substrate container with the first inlet port of the static mixing device, or the second sealable area is shaped to at least partially accommodate the conduit for fluidly connecting the outlet port of the second substrate container with the second inlet port of the static mixing device.

28. 28. The frame of any one of claims 1 to 27, further adapted to hold a Y-piece or T-piece, the Y-piece or T-piece being disposed within the conduit fluidly connecting the outlet port of the static mixing device with the inlet ports of the waste container and the product container, such that an inlet of the Y-piece or T-piece is fluidly connected with the outlet port of the static mixing device, a first outlet of the Y-piece or T-piece is fluidly connected with the inlet port of the waste container, and a second outlet of the Y-piece or T-piece is fluidly connected with the inlet port of the product container.

29. 29. The frame of claim 28, wherein a one-way stopcock valve is disposed in the conduit portion fluidly connecting the first outlet of the Y-piece or T-piece with the inlet port of the waste container and / or the conduit portion fluidly connecting the second outlet of the Y-piece or T-piece with the inlet port of the product container.

30. 30. The frame of claim 29, wherein the one-way stopcock valve comprises means for mechanically operating the valve, the means being oriented relative to or adapted to operate from the rear face of the frame.

31. 31. A frame as described in any one of claims 28 to 30, wherein a check valve is disposed in the conduit fluidly connecting the first outlet of the Y-piece or T-piece with the inlet port of the waste container, the check valve optionally being positioned downstream of the one-way stopcock valve.

32. The frame is adapted to be insertable in its operative orientation into an apparatus for aseptically mixing two fluids, the apparatus comprising: a first counterpiece for sealingly covering said first and said second sealable areas; means for applying pressure to said first and second flexible substrate containers when secured to or inserted into said first and second sealable areas, 32. The frame of any one of claims 1 to 31, wherein the frame optionally holds the first and second substrate containers, the flexible waste container, the flexible product container, the static mixing device, and the conduits for fluidly connecting the outlet port of the first substrate container with the first inlet port of the static mixing device, the outlet port of the second substrate container with the second inlet port of the static mixing device, and the outlet port of the static mixing device with the inlet ports of the waste container and the product container.

33. 33. The frame of claim 32, wherein the pressure is exerted by pressurized gas contacting the outer surfaces of the first flexible substrate container and the second flexible substrate container, and the pressurized gas is provided to the surfaces through the first through hole located in the first sealable area and the second through hole located in the second sealable area.

34. 34. A frame according to claim 32 or 33, wherein the rear face of the frame is adapted to be sealed against a second counterpiece by circumferential gaskets respectively surrounding the first and second through holes.

35. 35. The frame of claim 34, wherein the first counterpiece and the second counterpiece are hingedly connected to one another.

36. 36. A flexible container adapted for use as a flexible substrate container, flexible waste container, or flexible product container according to any one of claims 1 to 35, comprising: an interior space for holding a fluid material surrounded by a flexible front wall and a flexible rear wall, each wall made of a polymeric material; and at least one inlet or outlet port for allowing fluid communication with the interior space, wherein the flexible front wall and the flexible rear wall are connected to each other to form a sealing edge that substantially encloses the interior space, the edge including four corner regions such that when empty the interior space has an overall square or rectangular shape, wherein a first through-hole is provided at or near a first corner region of the sealing edge, and a second through-hole is provided at or near a second corner region of the sealing edge, the second corner region being adjacent to the first corner region.

37. 37. The flexible container of claim 36, comprising at least one inlet port and / or at least one outlet port, with flexible tubing fluidly connected to the inlet port and / or outlet port, the flexible tubing comprising a sterile disconnect.

38. 38. The flexible container of claim 37, wherein the flexible tubing fluidly connected to the at least one outlet port has a downstream end fluidly connected to a sampling tube, the sampling tube having a downstream end fluidly connected to a sterile filter.

39. A kit comprising a frame according to any one of claims 1 to 35, (a) the first and second flexible substrate containers; (b) the flexible waste container; and (c) the flexible product container; and (d) the static mixing device; (e) a conduit for fluidly connecting the outlet port of the first substrate container with the first inlet port of the static mixing device, the outlet port of the second substrate container with the second inlet port of the static mixing device, and the outlet port of the static mixing device with the inlet ports of the flexible waste container and the flexible product container, and optionally for filling each of the first and second flexible substrate containers with a liquid substrate.

40. the static mixing device comprises a T-piece mixer, a Y-piece mixer, a vortex mixer, a baffle-based static mixer, a microfluidic mixing device, a multi-inlet vortex mixer (MIVM), or a jet impingement reactor, the jet impingement reactor optionally comprising: a reaction chamber defined by the inner surface of a reaction chamber wall, said reaction chamber having a substantially spheroidal overall shape, said chamber comprising: (a) first and second fluid inlets, the first and second fluid inlets being disposed at opposing positions on a first central axis of the reaction chamber so as to face each other, and each of the first and second fluid inlets comprising a nozzle; (b) a fluid outlet disposed at a third location, the third location being located on a second central axis of the chamber, the second central axis being perpendicular to the first central axis; 40. The kit of claim 39, wherein a distance between the nozzle of the first fluid inlet and the nozzle of the second fluid inlet is equal to or less than a diameter of the reaction chamber along the first central axis.

41. The jet impingement reactor comprises a reaction chamber, the chamber having a substantially spherical shape, the spherical shape comprising: first and second fluid inlets, the first and second fluid inlets being disposed at opposing positions on a first central axis of the reaction chamber so as to face each other, and each of the first and second fluid inlets being provided by a nozzle; a fluid outlet disposed at a location located on a second central axis of the chamber, the second central axis being perpendicular to the first central axis; the reactor further comprising first, second, and third fluid conduits, the first and second fluid conduits being arranged to direct a first fluid to the first fluid inlet and a second fluid to the second fluid inlet, and the third fluid conduit being arranged to direct a third fluid downstream from the fluid outlet, the third fluid being formed by the mixing or reaction of the first and second fluids in the reaction chamber; 41. The kit of claim 39 or 40, wherein the reactor comprises at least two pieces secured together, a first piece of which is made of a polymeric material and comprises at least a portion of the first or second fluid conduit and at least a hemispherical portion of the reaction chamber, and a second piece of which is at least partially insertable into the first piece and comprises the fluid outlet.

42. 42. The kit of any one of claims 39 to 41, wherein the flexible substrate container and optionally the flexible waste container are containers according to claim 36, and the flexible product container is a container according to claim 37 or 38.

43. 36. An apparatus adapted to receive and hold in its operational orientation a frame according to any one of claims 1 to 35, wherein the first and second flexible substrate containers are empty and are held in the first and second sealable areas, the apparatus further comprising means for aseptically filling the first flexible substrate container with a first substrate and / or means for aseptically filling the second flexible substrate container with a second substrate.

44. 36. An apparatus adapted to receive and hold in its operative orientation a frame according to any one of claims 1 to 35, wherein the first flexible substrate container contains a first substrate and the second flexible substrate container contains a second substrate, the apparatus further comprising means for forcing the substrates to flow from the first and second flexible substrate containers into the static mixing device so as to mix and form a liquid product, the means being adapted to apply pressure to the first and second flexible substrate containers.

45. 45. The apparatus of claim 44, wherein the pressure is exerted by pressurized gas contacting the outer surfaces of the first and second flexible substrate containers, the pressurized gas being provided to the surfaces through the first through-hole located in the first sealable area and the second through-hole located in the second sealable area.

46. 35. The apparatus of claim 34, comprising the first counterpiece and the second counterpiece.

47. 47. The device of claim 46, wherein the first counterpiece and the second counterpiece are hingedly connected to one another.

48. 10. A method for mixing two fluid substrates, the method comprising the use of a frame according to any one of claims 1 to 35, a kit according to any one of claims 39 to 42, a flexible container according to any one of claims 36 to 38, and / or an apparatus according to any one of claims 43 to 47.

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