A portable air-blast system for homogeneous and reproducible freezing and thawing of biological materials

Abstract

The present disclosure relates to a system and method for improving the uniformity of thermal conductivity in a container filled with biological material during a freeze-thaw process, particularly a system that can be easily transferred and added to other conventional freeze-thaw methods and devices. The system includes a portable air blast system configured to receive a container filled with biological material and place it in a cooling or heating chamber for uniform and reproducible freezing and thawing of the biological material. The present disclosure also relates to a method for freezing and thawing a biological solution in a container using the portable air blast system of any of claims 1-12, the method including the steps of obtaining a container filled with biological solution; placing the container in a vent enclosure and on a stand for the air blast system; placing the air blast system together with the container in a heating or cooling chamber; and activating the air blast system by activating a fan in the air blast system.

Description

[Technical field]

[0001] This disclosure relates generally to systems and methods for homogeneously and reproducibly freezing and thawing biological materials, particularly aqueous solutions of biological materials used in chemical and pharmaceutical processes. This disclosure relates to systems and methods for improving the uniformity of thermal conductivity of a vessel filled with biological material during a freeze-thaw process, and in particular to a system that is easily transferable and additive to other conventional freeze-thaw methods and apparatus. [Background technology]

[0002] Biological materials are industrially produced in large batches that are stored for long periods of time and are generally frozen to minimize degradation during their relatively long storage period. The freezing and thawing process of biological solutions presents several challenges to consider, including the freeze-thaw operating parameters, storage of the material, transportation in a frozen state, and logistics. The choice of storage container is one of the most important considerations in the freeze-thaw process, as it can affect the stability of the formulation and downstream processing. Commonly used containers for frozen biological materials include bags and bottles / carboys. Although disposable bags have advantages over bottles / carboys, the risk of contamination and the potential risk of damage to the bag or tubing causing product loss are of greater concern than with bottles / carboys. The bottle / carboy system is simpler and more robust than bags and is the preferred embodiment for many companies, provided the biological product is robust and stable under a wide range of freeze-thaw conditions.

[0003] Currently, the freezing process involves placing the bottles and / or carboys containing the biological solution in a conventional upright or large freezer and allowing the product to freeze. However, product loss is often observed when freezing biological solutions using conventional freezers with set point temperatures between -20°C and -80°C. This is because such freezers are inadequate to provide the cooling rate required for a given load configuration. In conventional freezers, containers are placed side-by-side and sometimes stacked in the freezer, resulting in temperature gradients that are not uniform across the freezer space and differ from container to container. Thus, under these conditions, the freezing rate and product quality depend on the freezer capacity, freezer load, space between containers, container size, container shape and airflow characteristics within the freezer. Another option for the freezing process involves the use of cold air and blast freezers to rapidly freeze the biological material below its glass transition temperature. In a typical air blast freezer, cold air is circulated by a fan over containers that are enclosed in an insulated, sealed room or chamber. The degree of cooling achieved depends on several factors, including fan location, chamber volume, load configuration, temperature and air velocity, etc. Even with this rapid freezing method, inconsistencies in freezing from container to container can occur, which can result in inconsistencies in the quality of product batches.

[0004] Another consequence of different freezing rates is the occurrence of inhomogeneity of solute distribution on the macroscale in the frozen solution of biological materials (macro-cryoconcentration or freeze concentration). Cryoconcentration is associated with an inhomogeneous ice matrix, which may lead to degradation and quality loss of biological materials. Slow freezing and thawing also impairs the stability of biological materials. Furthermore, heat transfer at the top of the container, both by convection and radiation, leads to the formation of an ice shell consisting of an ice layer on top of the liquid at the gas-liquid interface. The ice shell increases the internal pressure in the container, leading to damage and destruction of the container. Patent document 1, filed on November 12, 2018, discloses an insulation device that prevents the formation of an ice shell on top of the liquid at the gas-liquid interface in the headspace region of the container. This device avoids the formation of an ice shell and the increase in pressure in the container, preventing damage and rupture of the container, thereby improving the freezing process.

[0005] Bottles and carboys are widely used by many pharmaceutical companies, but a scale-down model that mimics a large-scale system is desirable for research and development and optimization of the freezing and thawing process. Recently, Patent Document 2 discloses a scale-down system designed for bottles.

[0006] Although systems and methods already exist to help improve the freezing and thawing process using bottles and carboys, such as ice shell insulation devices and scale-down systems, these systems have yet to address the problem of inhomogeneity of freezing across multiple containers of a batch. Specifically, they have yet to address the problem of inhomogeneity of freezing when using currently available freezers, conventional or blast freezers, and freezers installed at different locations. Since in most cases the freezing equipment is already installed and varies across sites of the same biopharmaceutical company, it is desirable to design a simple and portable system for homogeneous and reproducible freezing and thawing of biological materials that can be used with existing equipment at different locations. Furthermore, it would be advantageous if the system and method could incorporate the ice shell insulation devices and scale-down systems already disclosed, thereby preventing the above-mentioned problems. This would allow for homogenization of the freezing process across containers, even when using different freezer types. [Prior art documents] [Patent documents]

[0007] [Patent Document 1] Portuguese Provisional Patent Application No. 115152 [Patent Document 2] International Publication No. 2018211437 Summary of the Invention [Problem to be solved by the invention]

[0008] The present disclosure provides systems and methods for uniform and reproducible freezing and thawing of biological materials, particularly systems that can be easily transported and used with other conventional freeze-thaw methods and devices.

[0009] The present disclosure provides systems and methods for improving the uniformity of thermal conductivity in a container filled with biological material during a freeze-thaw process. [Means for solving the problem]

[0010] The disclosed system includes a portable air blast system configured to receive a container filled with biological material and place it in a cooling or heating chamber. The system allows for uniform and reproducible freezing and thawing of the biological material. The portable air blast system improves the uniformity of thermal conductivity in the walls of the container, allowing for the same freezing / thawing rates even when different cooling or heating chambers are used.

[0011] The portable air blast system disclosed herein includes a stand for housing the container and for maintaining the container in a centered position within a vent enclosure. The portable air blast system also includes a vent enclosure designed to ensure uniform vertical air flow, and further includes a fan for drawing air within the chamber through the vent to improve uniformity of thermal conductivity at the container walls during the freeze / thaw process.

[0012] Another aspect of the present disclosure relates to a method of improving thermal conductivity uniformity in a container filled with biological material during a freeze-thaw process using the systems disclosed herein, the method including the steps of: (1) providing a cooling or heating chamber; (2) providing at least one portable air blast system; (3) placing the container in the center of the portable air blast system; (4) placing the portable air blast system within the cooling or heating chamber; and (5) running a fan until the biological material is completely frozen or thawed.

[0013] Another aspect of the present disclosure relates to rapidly loading a chamber into one or more portable air blast systems using multiple portable air blast systems and trolleys combined into one chamber.

[0014] Another aspect of the present disclosure relates to a platform configured to house a portable air blast system and capable of rotating, rocking, quivering, vibrating or providing other forms of mechanical motion to induce convection of liquid within a container, the platform being used during the thawing process.

[0015] One aspect of the present disclosure relates to a system for freezing and thawing an aqueous solution of a biological material, and in particular, a system for improving thermal conductivity uniformity in a container filled with the biological material, the system including a portable air blast system configured to house the container; the container filled with an aqueous solution of the biological material; and a cooling or heating chamber housing the portable air blast system for freezing or thawing.

[0016] In one embodiment, the portable air blast system is configured to house the container and is made of a rigid material such as a plastic, polymer, or other rigid material.

[0017] In one embodiment, the portable air blast system is configured to house the container and includes a stand, a vent enclosure, and a fan enclosure.

[0018] In one embodiment, the portable air blast system includes a stand that receives the container and maintains its position in the center of the vent enclosure.

[0019] In one embodiment, the stand may have a support designed to receive the container.

[0020] In one embodiment, the stand may have a pin that connects the stand to the vent enclosure.

[0021] In one embodiment, the pin maintains the distance between the stand and the vent enclosure at between 1 cm and 10 cm, more preferably between 2 cm and 5 cm.

[0022] In one embodiment, the portable air blast system is equipped with a vent enclosure to obtain uniform vertical airflow on all side walls of the container.

[0023] In one embodiment, the distance between the wall of the vent enclosure and the side wall of the container is approximately constant, between 1 cm and 10 cm, preferably between 1 cm and 3 cm.

[0024] In one embodiment, the velocity of the air within the vent is preferably greater than twice the downward velocity of the outside air.

[0025] In one embodiment, the air velocity within the vent is between about 0.5 m / s and about 20 m / s, preferably between about 1 m / s and about 10 m / s, and more preferably between about 2 m / s and about 8 m / s.

[0026] In one embodiment, the portable air blast system includes a fan enclosure having a fan for passing air from a chamber or room to a vent.

[0027] In one embodiment, the fan may comprise a support, preferably an insulating support, connected to the vent enclosure.

[0028] In one embodiment, the fan is located at the top of the vent enclosure, preferably above the vessel.

[0029] In one embodiment, a fan or blower may be used, preferably a fan, more preferably an axial fan.

[0030] In one embodiment, the fan is battery powered.

[0031] In one embodiment, the fan may preferably have a control system for controlling the speed.

[0032] In one embodiment, the fan must be suitable for use in cryogenic environments.

[0033] In one embodiment, multiple fans may be used, preferably arranged side-by-side, either horizontally or vertically.

[0034] In one embodiment, the portable air blast system may be made up of modular segments that are attached to one another.

[0035] In one embodiment, the container is a fixed shape container.

[0036] In one embodiment, the container is constructed from a rigid, biocompatible material such as glass, polyethylene terephthalate, polycarbonate, polytetrafluoroethylene, polyethylene, polyester, polyamide, polypropylene, ethylene-vinyl alcohol copolymers, polyvinylidene fluoride, polyvinyl chloride, and the like, as well as copolymers, blends, or laminates containing these.

[0037] In one embodiment, the container has a volume of about 1 mL to about 20 L, preferably about 100 mL to about 10 L.

[0038] In one embodiment, the container is a deformable container.

[0039] In one embodiment, the deformable container is composed of a biocompatible polymeric material, such as ethylene-vinyl acetate copolymer, ethylene-vinyl alcohol copolymer, polytetrafluoroethylene, polyethylene, polyester, polyamide, polypropylene, polyvinylidene fluoride, polyurethane, polyvinyl chloride, and copolymers, blends, and laminates thereof.

[0040] In one embodiment, the deformable container has a volume of about 10 mL to about 20 L, preferably about 10 mL to about 1 L.

[0041] In one embodiment, the deformable container is placed within a rigid shell, preferably made of a low heat resistant material such as metal (eg, stainless steel, aluminum or copper).

[0042] In one embodiment, the vessel may include an ice crust attenuation device.

[0043] In one embodiment, the vessel may be equipped with a scale-down device.

[0044] In one embodiment, the cooling or heating chamber may be a static or air blast chamber, or a controlled temperature chamber.

[0045] In one embodiment, multiple portable air blast systems may be grouped together in one chamber.

[0046] In one embodiment, there is a typical distance between the portable air blast systems to ensure that the air velocity outside the vent is between about 0.05 m / s and about 1 m / s, preferably between about 0.1 m / s and about 0.5 m / s.

[0047] In one embodiment, the trolley is used to load one or more portable air blast systems into the chamber.

[0048] In one embodiment, a platform configured to house the portable air blast system can rotate, rock, tremble, vibrate, or provide other forms of mechanically inducing convection currents in the liquid within the container.

[0049] One aspect of the present disclosure relates to a method of improving thermal conductivity uniformity in a container filled with biological material, the method comprising providing a cooling or heating chamber, providing at least one portable air blast system, centrally mounting the container on the portable air blast system, mounting the portable air blast system in the cooling or heating chamber, and running a fan until the biological material is completely frozen or thawed.

[0050] In one embodiment, biological materials include proteins, amino acid and peptide preparations, nucleic acid (DNA, RNA) solutions, cell suspensions, tissue suspensions, cell aggregate suspensions, cell growth media, serum, biological products, blood products, storage fluids, fermentation broths, and cell culture media with or without cells, mixtures of the above, and fragments thereof.

[0051] One aspect of the present disclosure relates to a portable air blast system for freezing and thawing a biological solution in a container, the system comprising: a vent enclosure having a wall; a stand configured to accommodate a container having a sidewall; and at least one fan, wherein the fan passes air through the vent enclosure to uniformly distribute heat within the wall of the container, and wherein a distance between the wall of the vent enclosure and the sidewall of the container is approximately constant to ensure similar vertical airflow velocity around all sidewalls of the container.

[0052] In one embodiment, the portable air blast system further comprises a controller for controlling the vertical air velocity.

[0053] In one embodiment, the portable air blast system further comprises a fan enclosure.

[0054] In one embodiment, the portable air blast system comprises a vent enclosure having a wall; a stand configured to house a container having a sidewall; and at least one fan, wherein the distance between the stand and the vent enclosure is between 1 cm and 10 cm, the fan passes air through the vent enclosure to ensure uniform thermal conductivity within the wall of the container, and the distance between the wall of the vent enclosure and the sidewall of the container is between 1 cm and 10 cm and is approximately constant to ensure similar vertical air velocity around all sidewalls of the container.

[0055] In one embodiment, the stand of the portable air blast system further comprises a solid base and a pin for connecting the stand to the vent enclosure.

[0056] In one embodiment, the stand of the portable air blast system is configured to maintain the position of the container in the center of the vent enclosure.

[0057] In one embodiment, the distance between the stand and the vent enclosure is 2 cm to 5 cm.

[0058] In one embodiment, the distance between the vent enclosure wall and the sidewall of the container is between 1 cm and 3 cm and is approximately constant to ensure similar vertical air velocity around all sidewalls of the container.

[0059] In one embodiment, the vertical air velocity is from about 0.5 m / s to about 20 m / s, preferably from about 1 m / s to about 10 m / s, and more preferably from about 2 m / s to about 8 m / s.

[0060] In one embodiment, the portable air blast system is made from a hard material, preferably a plastic, and more preferably a polymer.

[0061] In one embodiment, the fan is located in a fan enclosure.

[0062] In one embodiment, the fan is an axial fan.

[0063] One aspect of the present disclosure relates to a method of freezing and thawing a biological solution in a container using the disclosed portable air blast system, the method including the steps of obtaining a container filled with the biological solution; placing the container in a vent enclosure and on a stand of the air blast system; placing the air blast system together with the container in a heating or cooling chamber; and activating the air blast system by activating a fan in the air blast system.

[0064] In one embodiment, the method of freezing and thawing a biological solution in a container further comprises installing a plurality of air blast systems in the chamber.

[0065] In one embodiment of a method for freezing and thawing a biological solution in a container, the position of the container is maintained in the center of the vent enclosure.

[0066] In one embodiment of a method for freezing and thawing a biological solution in a container, each air blast system is placed a short distance apart to ensure that the air velocity outside the air blast system vents is between 0.05 m / s and 1 m / s, preferably between 0.1 m / s and 0.5 m / s.

[0067] These and other objects, features and advantages of the present disclosure will become apparent from the following detailed description taken in conjunction with the accompanying drawings.

[0068] To facilitate an understanding of the present disclosure, reference is made to the accompanying drawings, which illustrate preferred embodiments of the present disclosure and are not intended to limit the scope of the present application. [Brief description of the drawings]

[0069] [Figure 1] FIG. 1 shows the time-temperature profile inside a bottle during the freezing process using a blast freezer (solid line) or a static freezer (dashed line). [Figure 2a] FIG. 1 is a cross-sectional view of a bottle 50 frozen in an air blast freezer equipped with a sideways fan, showing the inhomogeneity of ice growth 501. [Figure 2b] FIG. 1 is a side view of a bottle 50 frozen in an air blast freezer equipped with a lateral fan, showing lateral deformation 502 of the bottle 50. [Diagram 3] FIG. 1 is a front view of a portable air blast system 10 constructed in accordance with the present disclosure. [Figure 4] 1 is a cross-sectional schematic diagram of a portable air blast system 10 including a container 50 and an ice crust attenuation device 60. [Diagram 5]1 is an exploded perspective view of a portable air blast system 10 including a stand 20, a vent enclosure 30 and a fan enclosure 40. The air blast system is constructed in accordance with the present disclosure. [Figure 6] FIG. 1 is a partially exploded elevational view of a portable air blast system 10 constructed in accordance with the present disclosure. [Figure 7] 1 is an exploded cross-sectional schematic diagram of a portable air blast system 10, a container 50 and an ice crust attenuation device 60. The air blast system is constructed in accordance with the present disclosure. [Figure 8] 7 is an exploded cross-sectional schematic diagram of a portable air blast system 10 with a deformable container 70 within a rigid shell 701. The air blast system is constructed in accordance with the present disclosure. [Figure 9] 1 is a time-temperature profile within a bottle during the freezing process in a static freezer with (solid line) and without (dashed line) the portable air blast system 10. [Figure 10a] FIG. 1 is a cross-sectional view of a bottle 50 being frozen in a static freezer using a portable air blast system 10, the air blast system constructed in accordance with the present disclosure. The cross-sectional view shows a flat ice surface 504. [Figure 10b] FIG. 5 is a cross-sectional view of a bottle 50 frozen in a static freezer, showing the formation of an ice crust and a "pyramid" shape 501. [Figure 11] FIG. 1 is a perspective view of a plurality of portable air blast systems 10 installed in a cooling or heating chamber 80. The air blast systems are constructed in accordance with the present disclosure. [Figure 12] FIG. 1 is an exploded cross-sectional view of a portable air blast system 10 designed into a modular assembly, constructed in accordance with the present disclosure. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0070] As mentioned above, the freezing and thawing process of biological materials poses several challenges. One of the main issues is the lack of uniformity associated with the freezing and thawing phenomenon. Many variables contribute to the uneven freezing, with the main issue being related to the different freezing rates of multiple containers within a single batch of product. Currently, the freezing process involves placing bottles and / or carboys containing biological materials into a traditional upright or box-type bulk freezer or blast freezer to freeze the product. The freezing rate and product quality depend on the freezer capacity, freezer load configuration, space between containers, container size, container shape and airflow characteristics within the freezer. Although the freezing time is shorter when using a blast freezer than when using a static freezer (Figure 1), freezing non-uniformity occurs, which is mainly due to the nature (direction and speed) of the airflow. Depending on the freezer load and configuration, this non-uniformity can be increased. In addition to the non-uniformity of freezing rate between containers, a very important aspect is the non-uniformity of ice growth 501 within the container 50 (Figure 2a). This inhomogeneity in freezing rate can lead to problems with cryoconcentration and deformation 502 or destruction of the container 50 due to uncontrolled ice growth and increased internal pressure (FIG. 2b). It is therefore important to provide a system and method for freezing and thawing biological material in a homogeneous and reproducible manner, ensuring controlled growth of ice and avoiding increased internal pressure.

[0071] Another issue concerns the existence of different freeze-thaw equipment used at several sites of the same biopharmaceutical company. Therefore, it is desirable to have a portable system that can be easily used in different locations to achieve a homogenous and reproducible freezing and thawing of biological materials and can be used with existing equipment. Furthermore, having a portable system allows the air blast system to be shipped together with any product, thus allowing precise thawing in other locations that may have less technical capacity. Thus, product quality and process reproducibility can be guaranteed throughout the supply chain, even when different equipment is used.

[0072] The present disclosure describes systems and methods that allow for improved uniformity of thermal conductivity on the exterior surface of a container filled with biological material during the freeze-thaw process while preventing damage or destruction of the container. The disclosed portable system allows any chamber to be used for freezing or thawing and can significantly reduce the time required to freeze or thaw biological material in a container.

[0073] In one embodiment, a system is configured to accommodate a container filled with biological material for freezing and thawing. The system includes a portable air blast system 10 configured to accommodate a container 50 filled with biological material. The portable air blast system 10 is configured to be installed in a cooling or heating chamber for uniform and reproducible freezing and thawing of the biological material. The main purpose of the portable air blast system is to improve the uniformity of thermal conductivity in the walls of the container, allowing similar freezing / thawing rates to be achieved using different cooling or heating chambers (see, e.g., Figures 3-8).

[0074] In one embodiment, the portable air blast system 10 includes a stand 20, a vent enclosure 30, and a fan enclosure 40 (see Figures 3-8). The portable air blast system is made of a rigid material such as a plastic, polymer, or other hard material.

[0075] In one embodiment, the stand 20 is designed to accommodate the container 50 and maintain the container in the center of the vent enclosure 30 to distribute air evenly across the sidewalls of the container. The stand 20 can have multiple supports 202 for accommodating the container 50, designed according to the container to be used with. The stand 20 may have a solid base 201 and pins 203 for connecting the stand 20 to the vent enclosure 30 and maintaining the distance between the stand 20 and the vent enclosure 30. The distance between the stand 20 and the vent enclosure 30 is 1 cm to 10 cm, preferably 2 cm to 5 cm, to ensure that the air is distributed to the vents 302. When the portable air blast system 10 is installed in an air blast chamber with lateral ventilation, the stand 20 has an opening at the bottom to ensure that the air is distributed evenly to the vents 302. In another embodiment, the container 50 may be maintained in the center of the vent enclosure 30 by support tabs connected to the vent enclosure 30 (see FIG. 5).

[0076] In one embodiment, the portable air blast system 10 includes a vent enclosure 30 designed to generate a vent 302 to obtain a nearly identical vertical velocity field around all side walls 503 of the container. To ensure a similar vertical velocity field around all side walls 503 of the container, the distance between the wall of the vent enclosure 301 and the side walls 503 of the container is nearly constant, between 1 cm and 10 cm, preferably between 1 cm and 3 cm. The vent enclosure 30 can separate the vertical air flow passing inside the vent 302 from the downward flow passing outside. The velocity of the air inside the vent 302 is preferably more than twice the downward velocity of the outside air. In a preferred embodiment, the velocity of the air inside the vent 302 is about 0.5 m / s to about 20 m / s, preferably about 1 m / s to about 10 m / s, more preferably about 2 m / s to about 8 m / s (see FIG. 7).

[0077] In one embodiment, the portable air blast system 10 includes a fan enclosure 40 having a fan 402 to pass air in the chamber or room through the vent 302 to improve the uniformity of thermal conductivity at the walls of the container during the freezing and thawing process. In a preferred embodiment, the fan 402 is placed on top of the vent enclosure 30 above the container with an insulating support 401. In one embodiment, a fan or blower is used, preferably the fan 402, more preferably an axial fan. In another embodiment, the fan can be powered with a battery, which can be placed in the insulating support 401. In another embodiment, for sensitive biological materials or scale down, the speed of the fan 402 can also be controlled to advantageously increase or decrease the thermal conductivity for different stages of the process. In one embodiment, the fan 402 is suitable for use in a cryogenic environment. In another embodiment, multiple fans 402 can be used to move air through the vent 302, for example, two or four fans positioned side-by-side horizontally or vertically, or one at the top and one at the bottom of the vent enclosure (see Figures 5-7).

[0078] In one embodiment, temperature probes are positioned at multiple points within the portable airblast system and within the container. The temperature probes indicate the temperature of the airflow at specific locations within the portable airblast system and indicate the time-temperature profile of the freeze / thaw of the biological material within the container. The temperature probes may include thermocouples, thermistors, or other conventional temperature sensing devices suitable for use in cryogenic environments.

[0079] In another embodiment, air velocity probes are placed at multiple points in the portable airblast system to provide information regarding air velocity within the portable airblast system at specific locations. The air velocity probes may be anemometers, pitot tubes, or other conventional sensing devices suitable for use in cryogenic environments.

[0080] In one embodiment, the container 50 configured to be filled with biological material can have several shapes and structural features, such as a bottle or carboy. Preferably, the container 50 should maintain its shape when empty and not deform significantly when filled with product. The container 50 can be made of a rigid, biocompatible material to enhance compatibility with biological materials. Examples of materials include glass, polyethylene terephthalate, polycarbonate, polytetrafluoroethylene, polyethylene, polyester, polyamide, polypropylene, ethylene-vinyl alcohol copolymer, polyvinylidene fluoride, polyvinyl chloride, and copolymers, blends, and laminates thereof. The size and capacity of the container 50 can vary. In a preferred embodiment, the capacity of the container 50 is about 1 mL to about 20 L, preferably 100 mL to about 10 L. The container 50 configured to be filled with biological material can include a headspace area and one cap having at least one port with a tube for aseptic filling and venting purposes.

[0081] In one embodiment, the container 50 also includes an ice crust attenuation device 60 configured to be attached to the headspace of the container. The primary purpose of the ice crust attenuation device 60 is to prevent the formation of ice crusts that could increase pressure within the container and result in damage.

[0082] In another embodiment, the vessel may also include a scaled-down apparatus that mimics the larger vessel for development studies and optimization of the freeze-thaw process. In this embodiment using a scaled-down apparatus, the air velocity in the vent can be conveniently adjusted to control the average thermal transfer rate to match, for example, the velocity of the larger vessel.

[0083] In another embodiment, the deformable container 70 may also be frozen or thawed in the portable air blast system 10, in which case the deformable container 70 is placed in a rigid shell 701. The rigid shell is preferably made of a material with low thermal resistance, such as a metal (e.g., stainless steel, aluminum, or copper). The deformable container 70, such as a bag, may deform when filled with the product and may be made of a biocompatible rigid polymeric material to promote compatibility with biological materials. Examples of biocompatible polymeric materials include ethylene-vinyl acetate copolymers, ethylene-vinyl alcohol copolymers, polytetrafluoroethylene, polyethylene, polyester, polyamide, polypropylene, polyvinylidene fluoride, polyurethane, polyvinyl chloride, and combinations thereof, copolymers, blends, laminates, and the like. The deformable container 70 may vary in size and volume. In a preferred embodiment, the volume of the deformable container 70 is about 10 mL to about 20 L, preferably about 10 mL to about 1 L (see FIG. 8).

[0084] In this embodiment, biological materials include proteins, amino acid and peptide preparations, nucleic acid (DNA, RNA) solutions, cell suspensions, tissue suspensions, cell aggregate suspensions, cell growth media, serum, biological products, blood products, storage fluids, fermentation broths, and cell culture media with or without cells, mixtures of the above, and fragments thereof.

[0085] The portable air blast system 10 shown in Figures 3-7 has particular applicability with general freezing or thawing processes when installed directly in the cavity of a chamber. The chambers mentioned above may or may not have convection and can be cooled or heated for freezing or thawing. The use of fans in the portable air blast system allows the air from the cooled or heated chamber to be directed parallel to the walls of the container without interference from other adjacent containers, ensuring that the freezing and thawing process of each container is not affected by the other. The system disclosed herein utilizes the existing cooled / heated air in the chambers, allowing any chamber to be used for freezing or thawing. Furthermore, the system disclosed herein is designed to be portable, lightweight and compact, preferably weighing less than 20 kg or not increasing the weight or size of the filled container by more than 100%. This transport can be easily done between sites and can be used in any chamber or room, ensuring homogenous, reproducible and faster freezing and thawing of biological materials.

[0086] Further examples of the use of the disclosed portable air blast system for freezing an aqueous solution in a container are detailed below.

[0087] In one embodiment, for example, the portable air blast system 10 was used to freeze 1.8 L of 5% (m / V) sucrose solution in a polyethylene terephthalate (PET) bottle with dimensions of 240(h)x120(w)x120(d) mm. Testing was performed using the ice shell attenuation device 60 described above. In one embodiment, the bottle was placed directly in a deep freezing chamber with a temperature setpoint of -80°C and frozen. In another experiment, the bottle was frozen in the portable air blast system and placed in a deep freezing chamber with a temperature setpoint of -80°C. Figure 9 shows the time-temperature profile in the bottle for both experiments with and without the portable air blast system 10. It was found that the freezing process was faster with the portable air blast system 10 (Figure 9, solid line). It took about 200 minutes to achieve a temperature of -30°C in the bottle with the portable air blast system, whereas it took about 435 minutes to reach the same temperature without the portable air blast system (Figure 9, dashed line). Further, as shown in Figure 10, the formation of ice crust on the liquid and "pyramid-like" shape 501 is avoided using the portable air blast system and ice crust attenuation device, resulting in a flat ice surface 504. Thus, these results show that the disclosed system enhances the freezing process and reduces the freezing time by improving the thermal conductivity in the container wall during the freezing process, while avoiding the formation of ice crust (Figure 10a), and consequently preventing damage to the container.

[0088] In another embodiment, multiple portable air blast systems 10 could be combined into one chamber 80 (see FIG. 11). In a preferred embodiment, each container has a portable air blast system with one fan on the container. In a preferred embodiment, there is a distance between the several portable air blast systems to ensure that the air velocity outside the vent is between about 0.05 m / s and about 1 m / s, preferably between about 0.1 m / s and about 0.5 m / s.

[0089] In another embodiment, the portable air blast system 10 is made up of modular segments that can be attached to each other. The portable air blast system 10 can have interchangeable modules for different functions and can accommodate different containers and devices. The modular assembly of the portable air blast system 10 allows for many possibilities to convert and adapt the system to different scenarios. By adding or modifying different modules, the system can be made larger in size to better fit different containers, change the airflow profile, and accommodate different devices such as probes, batteries, electronics, etc. The modular assembly allows for transportation, manufacturing, part replacement, and assembly in place (see FIG. 12).

[0090] In another embodiment, the system also includes a trolley to allow for rapid loading of the chambers onto the portable air blast system or onto multiple portable air blast systems.

[0091] In another embodiment, the container is agitated to mix the biological solution during thawing, which can be accomplished using a platform configured to house the portable air blast system and capable of rotating, rocking, quivering, vibrating or providing other forms of mechanical motion to induce convective currents of the liquid within the container.

[0092] Another aspect of the present disclosure relates to a method of improving the thermal conductivity uniformity of a container filled with biological material during a freezing or thawing process using the system described above, the method including the steps of: (1) providing a cooling or heating chamber; (2) providing at least one portable air blast system; (3) placing the container in the center of the portable air blast system; (4) placing the portable air blast system within the cooling or heating chamber; and (5) running a fan until the biological material is completely frozen or thawed.

[0093] The term "comprising" as used in this document is intended to indicate the presence of stated features, integers, steps or components, but does not exclude the presence or addition of one or more other features, integers, steps, components or groups thereof.

[0094] Those skilled in the art will appreciate that, unless otherwise indicated herein, the particular sequence of steps described is merely illustrative and may be modified without departing from the present disclosure. Thus, unless otherwise indicated, the steps described are unordered, i.e., where possible, the steps may be performed in any convenient or desirable order.

[0095] The present disclosure is in no way limited to the described embodiments, and those skilled in the art will recognize many possibilities for modifying it.

[0096] The above-described embodiments may be combined. The following claims further define certain embodiments of the present disclosure.

Claims

1. 1. A portable air blast system configured for freezing and thawing a biological solution in a container, comprising: a vent enclosure having a wall configured to create a vent within the portable air blast system; A stand having a plurality of supports; At least one fan; A controller; and Equipped with a fan enclosure, the stand and the plurality of supports house a container having a sidewall; the position of the container can be maintained in the center of the vent enclosure by support tabs protruding from the walls of the vent enclosure such that the distance between the walls of the vent enclosure and the side walls of the container is between 1 cm and 10 cm and is approximately constant to ensure similar vertical airflow velocities around all of the side walls of the container; the controller controls the vertical air velocity; the fan directs air through the vent to provide uniform thermal conductivity across the wall of the vessel; the vertical air velocity is between about 0.5 m / s and about 20 m / s; The portable air blast system, wherein the container is a bag, a bottle, or a carboy.

2. The portable air blast system of claim 1 , wherein the stand further comprises a solid base and a pin for connecting the stand to the vent enclosure.

3. The pin protrudes from the solid base to support a bottom of the vent enclosure, 3. The portable air blast system of claim 2, wherein the distance between the solid base of the stand and the bottom of the vent enclosure is between 2 cm and 5 cm.

4. 4. The portable air blast system of claim 1, wherein the distance between the wall of the vent enclosure and the side walls of the container is between 1 cm and 3 cm and is approximately constant to ensure similar vertical air velocity around all the side walls of the container.

5. The portable air blast system of any one of claims 1 to 4, wherein the vertical air velocity is between about 1 m / s and about 10 m / s.

6. The portable air blast system of any one of claims 1 to 5, wherein the system is made of a rigid material.

7. The portable air blast system of any one of claims 1 to 6, wherein the fan is disposed within the fan enclosure.

8. The portable air blast system of any one of claims 1 to 7, wherein the fan is an axial fan.

9. A method for freezing and thawing a biological solution in a container using a portable air blast system according to any one of claims 1 to 8, comprising the steps of: obtaining a container filled with a biological solution; placing the container within the vent enclosure and on the stand of the air blast system; placing the air blast system together with the container in a heating or cooling chamber; and The method includes activating the fan in the airblast system to activate the airblast system.

10. 10. The method of freezing and thawing a biological solution in a container as recited in claim 9, further comprising the step of installing a plurality of air blast systems in the chamber.

11. 10. The method of freezing and thawing a biological solution in a container according to claim 9, wherein the position of the container is maintained centrally in the vent enclosure.

12. 11. The method of freezing and thawing a biological solution in a container according to claim 10, wherein each airblast system is spaced apart from each other such that the speed of air outside the vent of the airblast system is between 0.05 m / s and 1 m / s.

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