Sterile connectable sensor patch

Abstract

A first connection unit having a plurality of sensor surfaces and adapted to be connected aseptically to a second connection unit, e.g. mounted on a flexible bioreactor bag.

Description

[0001] This application is a divisional application of the patent application with application number 201680008689.X, application date January 29, 2016, and invention title "Sterile Connectable Sensor Patch". Technical Field

[0002] This invention relates to sterile connectors, and more particularly to sterile connectors with sensors that can be used in cell culture. The invention also relates to methods for forming sterile connections. Background Technology

[0003] There is a trend toward single-use systems in biopharmaceutical manufacturing, particularly in cell culture operations. Typical single-use systems for cell culture include bioreactors in which cell cultures are contained in flexible bags, or expandable self-supporting bags that provide agitation by swaying (see, for example, U.S. Patent No. 6,190,913), or bags supported by stainless steel support structures, where agitation is typically provided by impellers (see, for example, U.S. Patent No. 7,629,167) or pneumatically (see, for example, WO 2007 / 068945A1).

[0004] In both systems, sensors that come into contact with the cell culture are required to monitor important variables such as cell density, pH, oxygen concentration, and the concentrations of various metabolites and / or nutrients. Such sensors are typically integrated into the bag via a welded port, as in EP2503320. However, many sensors cannot be sterilized using the same preferred method for sterilizing the bag.

[0005] Therefore, a convenient technology is needed to aseptically connect individually packaged sensors to single-use bioreactor bags. Summary of the Invention

[0006] One aspect of the present invention is to provide a connection unit for aseptic connection of a sensor to a bioreactor bag. This is achieved using a first connection unit as defined in claim 1.

[0007] One advantage is that the connecting unit can be individually packaged and sterilized using a different method than the bioreactor bag. A further advantage is that the sensor can be stored under refrigeration, eliminating the risk of damage to the sensor from folding the bag. A further advantage is that the number of ports on the bag can be minimized, and several different sensor combinations can be easily provided for a single bag type.

[0008] A second aspect of the invention is to provide a connection unit that is mounted on a bag and can be aseptically connected to a connection unit with a sensor. This is achieved using the second connection unit as defined in claim 1.

[0009] A third aspect of the invention is providing a sterile connection between the sensor and the bioreactor bag. This is achieved using the connection as defined in the claims.

[0010] A fourth aspect of the invention is to provide a method for aseptically connecting a sensor to a bioreactor bag. This is achieved using the method as defined in the claims.

[0011] Other suitable embodiments of the invention are described in the dependent claims. Attached Figure Description

[0012] Figure 1 The first connecting unit and the second connecting unit of the present invention are shown, a) before connection and b) after connection and clamping.

[0013] Figure 2 The first and second connecting units of the present invention are shown after being connected and clamped.

[0014] Figure 3 The first and second connecting units of the present invention are shown, a) after connection and clamping, and b) after the sensor moves toward the bag wall.

[0015] Figure 4 A bioreactor bag with a first connecting unit and a second connecting unit having the connection and clamping of the present invention is shown. Detailed Implementation

[0016] In one aspect, such as Figures 1-3As shown, the present invention discloses a first connecting unit 1 adapted to aseptically engage with a second connecting unit 2, wherein the first connecting unit includes a plurality of sensor surfaces 3. The sensors may be suitably located in a recess 4 or on the inner surface 5 of the recess, which is sealed to the surrounding environment before engagement and fluidly connected to the interior 29 of the bioreactor bag 30 via the second connecting unit after engagement. The first connecting unit may include a first terminal surface 6 having a first opening 7 of the recess 4, wherein the opening is sealed by at least a first releasably adhered membrane 8. The membrane may be arranged on the first connecting unit such that contact between the membrane and the first connecting unit and / or the recess is aseptic. The membrane may be held in place, for example, by an adhesive foam ring 9 applied to the terminal of the unit outside the opening 7. The membrane may also be suitably folded 180 degrees and, if desired, equipped with tabs 10 for easy removal. Furthermore, the first unit may have a gasket 11 mounted on the first terminal surface, adapted to seal adjacent to a similar gasket 12 mounted on a second terminal surface 13 of the second connecting unit 2. The first releasably adhered membrane 8 can be arranged to mate with a second releasably adhered membrane 14 disposed on a second terminal surface 13 of the second connecting unit having a second opening 15, and the first and second membranes can be adapted to be pulled out together in pairs after mating, such that the corresponding first opening 7 and second opening 15 in the first and second terminal surfaces aseptically mate. The first connector unit and the second connector unit can suitably have a circular cross-section. They can also each include flanges 21, 22 adapted to engage with clamp 23 to ensure connection after mating. The flanges can be adapted, for example, to use the well-known Tri-Clamp, also disclosed in ISO 2852. TM Or Tri-Clover TM Fixture engagement.

[0017] In some embodiments, the first connector unit includes at least two, such as at least three or at least four sensor surfaces. Having multiple sensor surfaces within the same connector unit is advantageous because only one port is required. Cell culture typically requires monitoring several variables and desires a minimum number of ports, as each port increases cost and may carry the risk of compromising sterility. One or more of these sensor surfaces (such as at least two, three, or four sensor surfaces) may be enzyme sensor surfaces, i.e., sensor surfaces containing immobilized enzymes, although one or more of the sensor surfaces may additionally or alternatively be electrochemical sensor surfaces or optical dye-based sensor surfaces. Enzyme sensors are commonly used, for example, to sense glucose (using immobilized glucose oxidase) and glutamine (typically using immobilized glutaminase) and may also be used to sense other nutrients / metabolites in cell culture. Enzyme sensors require refrigeration during storage and may be incompatible with the radiation sterilization methods commonly used for bioreactor bags. Therefore, individually packaged enzyme sensors need to be sterilely connected to the bag immediately before use. The first connector unit with the enzyme sensor may be sterilized, for example, using ethylene oxide, hydrogen peroxide, or other chemical sterilizing agents. By selecting a suitable sensor from the above group and combining it with a heat-resistant polymer, the sensor patch can also be sterilized by autoclaving or electron beam processing. Packaging it in a protective atmosphere (such as N2 or Ar) to prevent oxidation enhances the stability of enzyme- and dye-based sensors during sterilization and storage. This protective atmosphere is easier to achieve for individual sensor patches than for the entire bag.

[0018] In some embodiments, the ratio between the depth 16 and width 17 of the recess 4 is less than 1, such as less than 0.5 or less than 0.25. A lower aspect ratio is advantageous because it reduces the risk of cell blockage of the recess during culture. The risk of cell blockage can be further reduced by mounting the sensor surface on a movable part, such as... Figure 3 As shown in the diagram. The movable component can be, for example, an elastic diaphragm 18 or a bellows, which, after connection, can move toward the interior of the bag, for example, by the movement of the plunger 19 or by pneumatic action. The sensor surface 3 can then be moved to be substantially flush with or even protrude from the bag wall 20 into the interior 29 of the bag 30. Alternatively, the sensor surface can be mounted on a fixed component, such as... Figure 1 and Figure 2 As shown in the diagram. This provides a simpler design, eliminates the need for moving parts, and facilitates the construction of connections with low height. The fixing component can be used, for example, with a second connecting unit having a beveled edge, as discussed below.

[0019] In the second aspect, such as Figures 1-3As shown, the present invention discloses a second connecting unit 2 adapted to aseptically mate with the first connecting unit 1 disclosed above. The second connecting unit may be mounted (e.g., welded) as a port on a bag wall 20 and may include a second terminal surface 13 with an opening 15 in fluid communication with the interior of the bag. The opening may be sealed by a second releasably adhered membrane 14, such that contact between the membrane and the second connecting unit and / or the opening 15 is aseptic. The membrane may be secured in place, for example, by an adhesive foam ring 24 applied to the terminal surface 13 of the unit outside the opening 15. The membrane may also be suitably folded 180 degrees and, if desired, equipped with tabs 25 for easy removal. Furthermore, the second unit may have a gasket 12 mounted on the second terminal surface, adapted to seal adjacent to a similar gasket 11 mounted on the first terminal surface 6 of the first connecting unit 1. The second releasably adhered membrane 14 can be arranged to engage with the first releasably adhered membrane 8 arranged on the first terminal surface 6 of the first connecting unit having the first opening 7, and the first membrane and the second membrane can be adapted to be pulled out together after engagement, so that the corresponding first opening 7 and second opening 15 in the first terminal surface and the second terminal surface are aseptically engaged.

[0020] In some embodiments, the ratio between the depth 26 and the width 27 of the second opening 15 is less than 1, such as less than 0.5 or less than 0.25. A lower depth-to-width ratio is advantageous because it reduces the risk of cell blockage of the recess during culture. Alternatively, or additionally, the second opening may have, as Figure 2 The inclined edge 31 is shown. The inclined surface may, for example, make the second opening 15 have a generally truncated conical shape, wherein the base 28 of the truncated cone faces the internal volume 29 of the bioreactor bag 30. The ratio of the diameter of the base 28 of the truncated cone to the diameter of the top 35 (or the width 17 of the first opening in the first connector) may be at least 2.

[0021] In a third aspect, the present invention discloses a sterile connection formed by the cooperation of a first connecting unit 1 of any of the embodiments disclosed above and a second connecting unit 2 of any of the embodiments disclosed above. This connection may further include a clamp 23 holding the first and second units together. As discussed above, it may be, for example, a Tri-Clamp or Tri-Clover clamp. The clamp may engage the flanges 21, 22 of the first and second units. The sterile connection may have a total height h of less than 3 cm (e.g., less than 2 cm). This enables the use of connections on the bottom side of a swing-type bioreactor bag, such as... Figure 4 As is the case here. Placing the sensor on the bottom side is preferred because the bag will only be partially filled with liquid and the sensor must be in contact with the cell culture at least intermittently.

[0022] In a fourth aspect, the present invention discloses a method for forming a sterile connection, comprising the following steps: a) Provides the first connection unit as disclosed above and the second connection unit as disclosed above; b) The first unit and the second unit are fitted together such that the first opening and the second opening are aligned with each other and the first membrane and the second membrane are in contact with each other. c) Pull out the first membrane and the second membrane together, so that the corresponding first opening and second opening in the first terminal surface and the second terminal surface are sterilely matched.

[0023] In some embodiments, the method further includes step d) using a clamp on a mating first connecting unit and a second connecting unit to ensure a connection.

[0024] In some embodiments, the second connecting unit 2 is arranged on the flexible bioreactor bag 30, wherein the second opening 15 forms a port on the wall 20 of the flexible bioreactor bag, and the method further includes the step e of adding cell culture medium and cells to the bag.

[0025] In some embodiments, the method further includes step f) of culturing cells in a bag and monitoring at least two parameters using a sensor surface.

[0026] This written description uses examples to disclose the invention, including the best mode, and also enables those skilled in the art to practice the invention, including making and using any apparatus or system and performing any included methods. The patentable scope of the invention is defined by the claims and may include other examples that may occur to those skilled in the art. Such other examples are intended to be within the scope of the claims if they have structural elements that are not different from the literal language of the claims, or if they include equivalent structural elements that are not substantially different from the literal language of the claims. Any patent or patent application mentioned herein is hereby incorporated in its entirety by reference as if they were individually incorporated.

Claims

1. A sensor patch capable of aseptic connection, comprising: The first connecting unit includes: At least one sensor surface; A first terminal surface having a first opening of a first recess constructed for fluid flow therein, wherein the ratio between the depth and width of the first recess is less than 1, and wherein the first opening is sealed by a first releasably adhered membrane; and The first washer mounted on the surface of the first terminal; and A second connecting unit adapted to aseptically mate with the first connecting unit, comprising: A second releasably adhered membrane disposed on a second terminal surface, the second terminal surface having a second opening of a second recess configured for fluid flow therein, wherein the ratio between the depth and width of the second recess is less than 1, and wherein the second releasably adhered membrane is arranged to mate with the first releasably adhered membrane; and The second washer is mounted on the surface of the second terminal. The ratio between the depth and width of the first and second recesses is effective in reducing blockage of the fluid flowing therein. The first and second membranes are adapted to be pulled out together after mating, so that the corresponding first and second openings in the first and second terminal surfaces are aseptically mated, thereby forming a single recess from the first and second recesses, and the first and second gaskets are sealed adjacent to each other. Wherein, when the aseptically connectable sensor patch is fully assembled, the total volume of the individual recesses is equal to the sum of the first volume of the first recess and the second volume of the second recess; and The at least one sensor surface protrudes at least partially into the single recess.

2. The aseptically connectable sensor patch according to claim 1, wherein, The at least one sensor surface includes at least one enzyme sensor surface.

3. The aseptically connectable sensor patch according to claim 1 has a height of less than 3 cm.

4. The aseptically connectable sensor patch according to claim 1, wherein, The second opening has a truncated conical shape.

5. The aseptically connectable sensor patch according to claim 1 further includes clamps on the first connection unit and the second connection unit.

6. The aseptically connectable sensor patch according to claim 1, wherein, The second connecting unit is arranged on the flexible bioreactor bag, and the second opening forms a port in the wall of the flexible bioreactor bag.

7. The aseptically connectable sensor patch according to claim 1, wherein, The ratio between the depth and width of the first or second recess is less than 0.

5.

8. The aseptically connectable sensor patch according to claim 1, wherein, The ratio between the depth and width of the first or second recess is less than 0.

25.

9. The aseptically connectable sensor patch according to claim 1, wherein, The first releasably adhered membrane is itself folded 180 degrees and includes tabs on its edges for easy removal.

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