APPARATUS, KIT AND METHOD FOR THE SUPPLY AND USE OF ELECTROMAGNETIC FIELDS WITH RESPECT TO A BIOREACTION.

Abstract

The invention relates to an apparatus, a kit, and a method for improving the quality, accelerating a change, and / or enhancing the performance of a bioreaction mixture. The bioreaction mixture is typically contained within an interior space in a vessel, such as a biofabrication vessel, with means for moving the bioreaction mixture. One or more modules are provided to emit one or more electromagnetic fields to which the bioreaction mixture is exposed for a period of time.

Description

APPARATUS, KIT AND METHOD FOR THE SUPPLY AND USE OF ELECTROMAGNETIC FIELDS WITH RESPECT TO A BIOREACTION The invention to which this application refers is the application of one or more electromagnetic fields, such as a series of pulsed electromagnetic fields (PEMF), (which may also be called digital sequences of electromagnetism), to a material in the form of a bioreaction mixture to achieve an improvement with respect to the same, such as an improvement in performance, and / or an acceleration in the change of the condition of the material and / or an improvement in the quality thereof. Changes, improvements, and / or accelerations in conditions that can be influenced are particularly relevant in the bioreaction of a bioreaction mixture, and examples of which are the metabolic productivity of biosystems such as fermentation and cell culture biosystems. For the application of one or more electromagnetic fields to be effective on a relatively large scale, in terms of several liters of bioreaction mixture, it is preferable to be able to guarantee that one or more electromagnetic fields are applied reliably and repeatedly. This ensures that the beneficial effect is achieved substantially uniformly and that the entire bioreaction mixture is exposed to the emitted electromagnetic fields to a similar degree. It is also necessary that the effect and improvements be achieved in a repeatable and reliable manner, and that the process be controllable by untrained personnel. This maximizes the possibility of using the apparatus and method in relatively large-scale manufacturing environments, rather than being solely a laboratory-based process, as is conventionally the case with many bioreaction techniques. It is known how to deliver the bioreaction mixture within a vessel, and although known systems exist for applying a magnetic field to a bioreaction mixture, these systems typically include one or more coils located at the top and / or bottom of the vessel. Problems have been found with the ability of this type of apparatus to provide uniform treatment of the bioreaction mixture within the vessel and to achieve the required change in the condition of the bioreaction mixture within a commercially acceptable timeframe. The applicant, in his pending application WO2019 / 234442, the contents of which are incorporated herein by reference, described another form of apparatus. QL AAΩ / 77Π7 / E / YILI Therefore, one objective of the present invention is to provide a solution to the aforementioned problems and to provide an apparatus and method that is reliable and allows for relatively uniform treatment of a bioreaction mixture, thereby enabling improvements in the quality, performance, and / or rate of change of the bioreaction mixture compared to conventional apparatus and methods. Another objective is to enable this to be achieved in a reliable and uniform manner and on a scale suitable for commercial batch production. Another objective is to provide a non-invasive device and method for bioreaction mixing. Another objective of the present invention is to provide an apparatus and a method that allows the effective application of one or more electromagnetic fields to the bioreaction mixture in a manner that is relatively easy and reliably repeatable and that can preferably be used in conjunction with a bioreactor-shaped vessel in which the material is contained. In a first aspect of the invention, an apparatus is provided for improving the performance and / or acceleration and / or quality of a bioreaction, the apparatus comprising: a container formed by a plurality of walls comprising at least one side wall and a base forming an internal space for containing a bioreaction mixture, means for causing the movement of said bioreaction mixture, and one or more modules, said module comprising a housing and a transmitter disposed within said housing, said transmitter being capable of emitting one or more electromagnetic fields, wherein the one or more modules are close to at least a portion of said walls that is made of a material that is substantially transparent to said one or more electromagnetic fields, and / or wherein the one or more modules are disposed in said internal space. In one embodiment, one or more modules are arranged in the bioreaction mixture within the internal space. In one embodiment, the housing comprises a liquid-tight coating made of plastic or glass. In one embodiment, said plurality of walls includes a lid for the container. In one embodiment, said cover includes a portion made of a material that is substantially transparent to said one or more electromagnetic fields and said one or more modules are positioned close to them. Q ίββΩΩ / 77Ω7 / Β / ΥΙΛΙ In one embodiment, at least a portion of said material made substantially transparent to said one or more electromagnetic fields is made of a non-metallic material, such as plastic or glass. In one embodiment, the means for inducing motion are electrical and / or mechanical motion means. In one embodiment, the motion means take the form of any, or any combination of: an agitator within the internal space of the vessel; an agitator external to the internal space of the vessel; a means for imparting vibration to the vessel and / or bioreaction mixture; a wave generator; a flow of material entering the internal space; and / or a flow of material exiting the internal space. In one embodiment, one or more of these modules are included, joined or integral with said agitator. In one embodiment, the agitator within the internal space is an agitator, a propeller, an impeller and / or a gas sprayer. In one embodiment, these means of causing movement include one or more components of the bioreaction mixture. In one embodiment, the means for inducing movement move the bioreaction mixture to periodically expose substantially all of the bioreaction mixture to one or more electromagnetic fields emitted from one or more modules and / or to retain cells in the bioreaction mixture substantially suspended within it. This ensures that, although the emitted electromagnetic fields do not extend throughout the entire bioreaction mixture, the bioreaction mixture will move through the electromagnetic field emission zones and thus provide exposure of the entire bioreaction mixture to the electromagnetic fields over time. In one embodiment, the device includes a frame, and the container is formed by a bag made of an elastic and flexible plastic material and is supported by the frame. Typically, in this embodiment, one or more modules are attached to the frame and / or the sheet material. In one mode, these electromagnetic fields emitted by the transmitter are a series of pulsed electromagnetic fields (PEMFS). In one embodiment, the apparatus further comprises a control means for controlling the shape of said one or more electromagnetic fields emitted from one or more modules. In one modality, one or more electromagnetic fields are emitted at a frequency that causes the rotation of the molecular dipole of the water contained in the bioreaction mixture. QL AAΩ / 77Π7 / E / YILI In one mode, the control means control the emission of one or more electromagnetic fields at a frequency or frequencies in the range of 2.4 GHz to 2.5 GHz. In one mode, electromagnetic fields are emitted at a plurality of different frequencies in a random sequence during the emission time period of one or more electromagnetic fields. In one mode, when there are two or more modules, the one or more electromagnetic fields emitted by them are emitted in a synchronized manner. Typically, the control means are an integral part of the module, vessel, or frame. In one embodiment, the device includes a support and said one or more modules are located on said support and said support can be located with the container. In one configuration, one or more modules include a rechargeable power source. In one mode, this transmitter provides short-range wireless communication of electromagnetic fields. In one embodiment, the apparatus comprises one or more locators for locating one or more modules near, on, or within one or more walls of the container. Typically, the one or more locators comprise one or more of a hook, a sail clip, an adhesive patch, a threaded locator, a magnet, or any combination thereof. In one embodiment, when one or more modules are arranged in the internal space and in the bioreaction mixture contained therein, the one or more modules further comprise at least one flotation portion to contain the one or more modules at a depth in the bioreaction mixture, when the bioreaction mixture comprises a liquid. In one embodiment, when there are two or more modules, at least one of the modules has a flotation portion of a shape selected to retain it at a different depth in the bioreaction mixture than another module or modules in the bioreaction mixture when the bioreaction mixture is a liquid. In one embodiment, each of the two or more modules in a group of modules is provided with a flotation portion of a shape selected to retain said modules at different respective depths in the bioreaction mixture when the bioreaction mixture comprises a liquid. In one embodiment, the container includes one or more hollow elongated members extending from at least one wall, the base, and / or the lid such that the one or more elongated members are not in fluid communication with the internal space, and wherein at least one of the one or more QL AAΩΠ / 77Π7 / E / YILI hollow elongated members comprise at least one portion made of a material that is transparent to electromagnetic fields. In one modality, the one or more modules are two or more modules, and each of the two or more modules is located in a node of another module. In one mode, these adjacent modules are separated by a distance of half the wavelength of the frequency of the emitted electromagnetic fields. In another mode, these modules are located in the array such that there is a separation within a range of 5 to 10 cm between adjacent modules in the array. In one example, the wavelength of an electromagnetic field at a frequency of 2.45 GHz is approximately 12.2 cm, and therefore the modules are placed 6.1 cm apart. The wavelength paths intersect at the module locations and prevent interference between the modules, so these spacings can be used when positioning modules in a module array. Therefore, an apparatus is provided for applying one or more electromagnetic fields to a bioreaction mixture for a period of time by using at least one container in which said bioreaction mixture is held and at least one transmitter to emit one or more of said electromagnetic fields from thereto into one or more zones of the internal space of the container and into and through which said one or more zones the bioreaction mixture is moved by means of movement to ensure that substantially all of the bioreaction mixture is exposed substantially uniformly to the electromagnetic fields in the one or more zones. Typically, the transmitter is provided as part of a module and one or more modules may be located at least partially in said bioreaction mixture and / or near one or more walls of said vessel during the emission of said electromagnetic fields and said modules are controlled to emit said electromagnetic fields at a frequency, or a range of frequencies, continuously, or in pulses, during said period of time. Typically, electromagnetic fields are emitted at a plurality of different frequencies in a random sequence during a given time period. In one mode, these frequencies are in the 2.4–2.5 GHz range. In one mode, the specific frequencies are 2402 MHz, 2426 MHz, and 2480 MHz. In one embodiment, said housing includes location means to allow the module to be located in a selected location on, or adjacent to, one or more walls of said vessel or the module may be provided integral with a wall of said vessel or in a cavity or opening therein. In one scenario, the module or modules are provided along with at least part of the container as a kit. Once the kit has been used, the module or modules are refilled, if necessary, and reused. Alternatively, one or more modules can be recycled, for example, by returning them to the supplier, who can then charge for them and make the modules available for subsequent use, or for another user. Thus, the end user of the module or modules can be different from the supplier of said module or modules. In one embodiment, a support is provided for a plurality of said modules that allows said modules to be located with the container and retained in a predefined spaced array. Typically, a power source is provided to allow the components to receive sufficient power to operate for at least a predetermined period of time, and switching means are provided to allow selective operation of the module. In one mode, the power source is a rechargeable source provided within the module, and the connection to a charging facility is provided wirelessly or via a plug located on the module, and more typically when the module is not in use. In one modality, the specific frequency range of said one or more electromagnetic fields is the Industrial, Scientific and Medical (ISM) short-range radio frequency band. In one mode, the transmitter is capable of generating the electromagnetic field up to a distance of 15 meters. In one modality, the control means allow the frequency and digital sequence of the emitted electromagnetic fields to correspond to the dielectric and / or other properties of the water molecules within said bioreaction mixture. In one mode, the transmission of one or more electromagnetic fields is in millisecond pulses in the range of 2.4 - 2.5 GHz at low pulse frequencies between 10 and 20 Hz, so that the duty cycle is typically in the range of 1 to 2%. In one embodiment, the apparatus includes means of locating the nutrients required to be used in relation to and / or added to the bioreaction mixture in the interior space of the container. Typically, one or more modules are located and controlled to create a substantially uniform emission intensity of electromagnetic fields toward and within said bioreaction mixture. QL AAΩ / 77Π7 / E / YILI In one mode, the number of modules, the configuration of the array of the same and / or the intensity of the electromagnetic fields emitted by them will be selected to suit the particular use of the modules and / or the bioreaction mixture located in the container at a particular time of use. In another modality, the location of the modules on the wall of the container is selected with respect to the location of the bioreaction mixture and / or nutrients within the interior space of the container to, for example, allow the modules to be located in close proximity to the nutrients and / or bioreaction mixture by placing them on the outer surface of the wall opposite the location of the nutrients or bioreaction mixture. In one embodiment, the matrix extends from the base of the vessel upwards to a fill level where the bioreaction mixture can be present in the interior space of the vessel. In one embodiment, the matrix extends along a single axis, such as a column or a row, or is provided to extend along a plurality of axes. Typically, the modules are preset with respect to the particular frequency or frequencies of the electromagnetic field that are emitted and typically also with a predetermined pulse frequency. In one configuration, the module is supplied as a sealed unit, and access to the internal components within the housing is limited to connecting a charging cable for the module's power supply. In an alternative configuration, no charging cable is provided, and the power supply is charged wirelessly. In one mode, a wireless receiver provided in the module receives a change in the module's operating control parameters. In one mode, each module has a unique identification code. In one mode, the use of the module is disabled until a payment has been made to the module provider, said payment made by a customer and which will then allow the module to be used, typically for a series of on / off functions or for a specific period of time. In one mode, the module's control means control the frequency and digital sequence of one or more electromagnetic field signals that are emitted so that they correspond to and / or are related to the dielectric and / or other properties of the bioreaction mixture that is contained in the vessel at that time. QL AAΩ / 77Π7 / E / YILI In one modality, the control means are provided in the form of an integrated circuit provided in the housing and which includes the transmitter to allow the emission of electromagnetic fields from it. In one configuration, the device includes a charging and / or control unit that provides one or more modules, when not in use, with an electrical and / or data connection to enable activation and / or charging. In another configuration, the user can selectively activate their modules via a remote payment scheme. Once payment is received, the control unit switches one, some, or all modules—depending on the payment amount—to an active mode, enabling them to emit electromagnetic fields. Typically, the selective localization of modules in an array, together with the stirring means, allows substantially uniform exposure to electromagnetic fields transmitted from the modules over a predetermined period of time when the type of bioreaction mixture, vessel dimensions, amount of bioreaction mixture, and array configuration of the modules are selected accordingly. Typically, when a plurality of modules are provided in an array, the modules, in combination, act as a unitary electromagnetic field signal emitting medium. In one modality, the control means allow the synchronized and sequential emission of pulsed electromagnetic fields (PEMFS). In one mode, one or more electromagnetic field signals are emitted from the array of modules for all or part of the processing of the bioreaction mixture contained within the internal space of the vessel. In one modality, electromagnetic fields are emitted for a predetermined period of time that is determined with reference to the particular bioreaction mixture and / or the amount of the bioreaction mixture. In one mode, the application of one or more electromagnetic fields lasts from 30 minutes to 2 hours and can be performed simultaneously with another function, if required, such as cooling the bioreaction mixture. It should also be noted that the duration of pulsed electromagnetic field application depends on the type and / or quantity of the bioreaction mixture being treated and / or the desired end product. When electromagnetic fields are pulsed, providing a rest period between pulses ensures that the microorganisms in the bioreaction mixture are not overwhelmed by the 8 Q LRRnn / ZZnZ / E / YIAI electromagnetic energy, but rather encourages them to increase metabolic processes and increase the growth rate. This has been found to result in increased metabolite expression and more efficient nutrient conversion, thereby increasing yields and / or reducing the production time required to achieve the desired result. Furthermore, the rest periods between pulses can allow the activity generated in the bioreaction mixture to relax and promote homogeneity of the bioreaction mixture as the groups in the mixture break down and a thermodynamically favorable open structure of the bioreaction mixture naturally forms. In one embodiment, the use of the apparatus according to the invention provides any, or any combination, of increased productivity in the production of biofuels, cell cultures and genetically modified organisms, insulin, monoclonal antibodies, growth hormones, interferon, interleukins, blood factor VIIa, blood factor VIII, blood factor IX, erythropoietin, gonadotropin, glucagon, vaccine antigenic sequences, and mammalian cell culture. Typically, the microbial organisms in the bioreaction mixture are electromagnetic systems and respond to changes in electromagnetism. In one approach, electronic magnetic field detectors can be used in the vicinity of the apparatus to detect the generation of electromagnetic fields. In one modality, the use of electromagnetic fields is controlled for use under aerobic conditions, and in another modality, electromagnetic fields are applied to a bioreaction mixture in an aerobic environment. Although not the only use, the use of the device and method under aerobic conditions is expected to be particularly beneficial. In another aspect of the invention, a kit is provided for improving the performance and / or acceleration and / or quality of a bioreaction, the kit comprising: a container formed by a plurality of walls comprising at least one side wall and a base forming an internal space for containing a bioreaction mixture, a means for moving said bioreaction mixture, and one or more modules, said module comprising a housing and a transmitter disposed within said housing, said transmitter being capable of emitting one or more electromagnetic fields, wherein localization means are provided to allow the one or more modules to be positioned close to at least a portion of said walls made of a material that is substantially transparent to said one or more electromagnetic fields, and / or wherein the one or more modules are disposed in said internal space. QL AAΩ / 77Π7 / E / YILI In one embodiment, the container consists of a frame and a sheet material supported on the frame to form said interior space. In one embodiment, said kit includes a plurality of modules and said modules are adapted and / or can be located to allow said modules to be placed in a spaced array with respect to said bioreaction mixture. In one modality, the kit includes stirring means to move the bioreaction mixture during the transmission of one or more electromagnetic fields. In another aspect of the invention, a module is provided for emitting one or more electromagnetic fields, said module comprising a housing, a power source, a transmitter for emitting one or more electromagnetic fields, and control means for enabling the control of said one or more electromagnetic fields, wherein the one or more electromagnetic fields are controllable such that for a period of time they are emitted at a predetermined frequency or a predetermined frequency range and are emitted continuously or as a series of pulses with a spacing between said emission pulses, and said module is locatable in, or near, a bioreaction mixture such that at least part of the bioreaction mixture is within the range of the one or more emitted electromagnetic fields. In another aspect of the invention, a method is provided for improving the performance and / or acceleration and / or quality of a bioreaction, the method comprising the steps of: providing a container formed by a plurality of walls comprising at least one side wall and a base forming an internal space for containing a bioreaction mixture, moving said bioreaction mixture in said internal space, and providing one or more modules, said module comprising a housing and a transmitter disposed within said housing, said transmitter being capable of emitting one or more electromagnetic fields, emitting said one or more electromagnetic fields from said one or more modules into at least a portion of the internal space;and wherein one or more modules are close to at least a portion of said walls made of a material that is substantially transparent to said one or more electromagnetic fields, and / or wherein one or more modules are arranged in said internal space.; In one embodiment, the method includes the step of introducing a cell culture medium into the internal space of the container and introducing one or more cells into the cell culture medium to form a bioreaction mixture. In one embodiment, the cell culture medium includes water molecules. Typically, one or more electromagnetic fields are emitted at a frequency to rotate and modulate these water molecules. QL AAΩ / 77Π7 / B / YILI In one modality, the bioreaction mixture moves during the application of one or more electromagnetic fields. In one modality, the method includes the additional step of applying one or more electromagnetic fields to the cell culture medium before introducing one or more cells into it. In one modality, the emission of one or more electromagnetic fields includes applying pulsed electromagnetic fields to the bioreaction mixture. In one mode, one or more electromagnetic fields have a frequency between 2.4 and 2.5 GHz and / or are emitted in pulses in the range of 0.5-1.5 milliseconds (ms) in duration and / or such pulses are separated by rest periods that are in the range of 40-66 ms and / or the pulses are emitted within a range of 12-20 pulses per second. In one modality, the one or more cells are yeast cells and the bioreaction is for the production of bioethanol. In one modality, the one or more cells are mammalian cells and the bioreaction is for the production of nucleic acids or peptides. In one modality, one or more cells are hybridoma cells. In one modality, the one or more cells are insect cells and the bioreaction is for the production of nucleic acids or peptides. In one modality, a plurality of modules are provided and located in a matrix. In one mode, the use of at least one module is disabled until a proposed user of the module has made a payment to a module provider. In one modality, the payment made is equated to a series of on / off functions of the module and / or for a specific period of time of use of said module. In one mode, the module user selectively activates the module through a remote payment scheme, and when payment has been made, the control means change the module's condition, depending on the level of payment made, to an active mode in which one or more electromagnetic fields can be emitted from there. In one modality, the emission time period of one or more electromagnetic fields is equivalent to all or part of the time during which said bioreaction mixture is maintained in the interior space. In one mode, electromagnetic fields are emitted in pulses. QL AAΩ / 77Π7 / E / YILI The specific embodiments of the invention are now described with reference to the accompanying drawings; where Figures a-le illustrate a modality of a module according to a modality of the invention; Figure 2 illustrates a charging and / or control station or bank according to one embodiment of the invention; Figures 3a-3c illustrate ways of using the modules in Figures 1a-1e together with a bioreactor vessel; Figures 4 and 5 illustrate the rotation of the water dipole when exposed to the electromagnetic fields emitted according to the invention; Figures 6a-6d illustrate the apparatus and the results of experiments performed according to the invention using a Chinese hamster ovary (CHO) cell line and with respect to IgG production; Figures 7a and 7b illustrate another embodiment of the invention; and Figures 8a-8c graphically illustrate the results obtained from an additional set of experiments using a Murine Hybridoma Cell (MHC) line and with respect to IgG production. With reference first to Figures 1a-1e, a module 2 according to one embodiment of the invention is illustrated. In this embodiment, the module 2 includes a first and a second part 4, 6 which, when joined as indicated by the arrows 8 in Figure 1d, form an outer housing 10 that typically has a waterproof and dustproof seal. The housing may be provided with suitable sealing means depending on its particular format and intended use. In the embodiment shown, there is an opening 12 in part 4 for locating a button 14 that can be pressed to operate an on / off switch 16 to activate or deactivate the module.An additional opening or zone 22 may be provided to indicate the location of electromagnetic field emission and / or to allow enhanced emission of electromagnetic fields from the transmitter of module 20 provided on the printed circuit board 18 inside the housing. Typically, one transmitter will be provided per housing and module, but it should be noted that, in other configurations, a housing may have several transmitters located within it. Printed circuit boards 18, 24, 26 are all located inside the housing when formed and in this mode are held in place by means provided in Part 6 Q ΩΩΩΩ / 77Ω7 / B / YILI as shown in Figure 11. The main PCB 24 includes the control components for operating module 2 to control the use of the energy cell 28 and the frequency(ies) and continuous or pulsed emission of electromagnetic fields for a required period of time. The electromagnetic fields emitted in terms of frequency and / or continuous or pulsed electromagnetic fields may vary depending on the specific use of the modules at any given time. In one configuration, the module may include a receiver to allow the reception of control signals to update and / or alter the operation of the module and / or charge module 2. In one mode, the charging of one or more energy cells located within the housing can be carried out at a central location to which the modules are returned after use or, alternatively, it can be carried out at a remote location, such as the end user's location via a station 31, an example of which is illustrated in Figure 2. In this mode, two location units 30, 32 are shown, which are stackable via the support legs 34. In this configuration, each unit has a series of locations, in this case 1-6, each for the placement of a module, and two of the modules 2, 2' are shown in position. Each unit is provided for connection 36 to a power supply, and each location 1-6 is equipped with a wireless charging facility so that when the module is in position, the power source 28 within it is charged wirelessly. Additionally, each location can be provided with a means to enable data communication with the control media components within the module to allow the module's control media to be updated. In one configuration, the update may be to a new version of the control software and / or to enable the module to be usable.In the latter case, activation can be achieved through payment by the user, perhaps via the Internet or an application, or by another means, and the payment allows a signal to be sent to the units to allow one, some, or all of the modules to become active and usable for a predetermined time or number of uses. Figures 3a and 3b illustrate two embodiments of the apparatus in use. In both embodiments, a bioreactor vessel 38 is schematically shown, which, in these embodiments, includes a support frame 40 shown with dashed lines, and includes a means for moving a bioreaction mixture within an interior space of the vessel. In this embodiment, the means for moving is a stirrer 42, also shown with dashed lines, which is 13 QL AAΩ / 77Π7 / B / YILI provides a mechanism for rotation around axis 44 via motor 46. The vessel walls are formed by a bag 48 made of plastic sheet material, typically single-use, supported by the frame. The bag has multiple walls, including side walls 52 and a base 54, defining an interior space 50 that contains the bioreaction mixture. The bioreaction mixture 56 is provided within the interior space 50 at a level 58 and can be moved around the space by the agitator 42. Additional nutrients and / or other materials can be selectively added to the bioreaction mixture 56 during the process. Also provided according to the invention are a plurality of modules 2, and in the two embodiments shown, the plurality of modules 2 are provided so that, collectively, they can emit one or more electromagnetic fields 60 through at least one wall, in the embodiments shown, the side wall 52 of the bag 48 and into the bioreaction mixture 56 to at least create electromagnetic field zones through which the bioreaction mixture can move, and move for a period of time that is at least part of the total processing time for the bioreaction mixture. As illustrated in Figure 3c, with a module 2 shown as located, in this embodiment, by an adhesive layer 62 to the outer surface 64 of the side wall 52. Alternatively, or in addition, mechanical locating means are used to position the module. As an alternative to locating the module directly on the wall of the container, the module can be located at a distance from the wall, but close enough so that the one or more electromagnetic fields emitted by it can pass through the wall and enter the interior space to allow exposure of the bioreaction mixture to it. In Figure 3a, the modules 2 are provided in a column-like array configuration along axis 64 and typically at a height that coincides with the top level 58 of the bioreaction mixture 56 in the interior space 50. In Figure 3b, the array configuration provides a series of modules located along the horizontal axes 66 and the vertical axes 68, and it will be appreciated that the number of modules, the array configuration thereof, and the operating parameters thereof will typically be predetermined with respect to the bioreaction mixture to be processed, the amount of the bioreaction mixture, the size of the reactor vessel, and / or the required processing to be carried out. Q ίΩΩΩΩ / 77Ω7 / Β / ΥΙΛΙ In any modality, once processing is complete, the modules can be removed and then immediately reused or passed on for recycling and / or recharging and / or reprogramming, as appropriate.Typically, electromagnetic fields are emitted in the 2.4–2.5 GHz range. This electronic frequency, when emitted into a liquid bioreaction mixture containing water (H₂O), which has a positive molecular dipole (Δ1) and a negative molecular dipole (Δ3) as shown in Figure 4, rotates the dipoles (Δ1, Δ3), as illustrated in Figure 5. As at least some of the water molecules rotate (Δ7), typically one revolution per 2.4–2.5 GHz cycle, hydrogen bonds (H) break and reform, creating a perturbation wave around the hydrated surfaces of the bioreaction mixture being treated, such as cell membranes. This results in, among other effects, increased fluidity around the membrane and an improved interface with the aqueous medium, thus enabling improved process quality, yield, and / or acceleration.The emission of electromagnetic fields in this frequency range causes the modulation of the electric field component of the electromagnetic wave 55 and the adaptation of the frequency range of one or more electromagnetic fields so that the hydrogen bond breaking time corresponds to half a cycle of the frequency in the 2.4–2.5 GHz range of water molecules. The rate of change can also be affected by the temperature of the bioreaction mixture. In tests, the apparatus and method have been applied to the use of yeast ethanol expression and have a radically beneficial effect on yeast cells (eukaryotic cells). Pressure transducers convert the CO2 evolution into relative quantities of ethanol, from which it is clear that the emission of electromagnetic fields using the modules as defined herein not only doubles the rate of ethanol production but could also increase the total amount of ethanol when the process is allowed to run to completion. Other uses include improvements in the efficiency of nucleic acid transfection, improved performance of IgG antibodies in mammalian cell lines, industrial application in single-use bioreactors (by using the beneficial nature of plastic film material coatings that are transparent to electromagnetic fields), improved production of therapeutic proteins from yeast, and applications using modules with conventional bioreactors. Q LRAnn / ZZnZ / E / YIAI In the tests performed using a module of the type shown in Figures 11a, a Corning® disposable rotating flask bioreactor vessel, P / N: CLS3152, with a volume of 125 mL, provided by Sigma Aldrich, USA, was used. The vessel 70 is illustrated in Figure 6a and has an interior space 72, containing a stirrer 74, a base 76, a top or lid 78, side walls 80, and connecting channels 82, 84. The bioreaction mixture included cell culture and was located within the interior space 72 in the experiments. More specifically, the material in this set of experiments was a CHO cell line producing monoclonal IgG, and the experiments studied the effects of using a module and emitting its electromagnetic fields on antibody production from the bioreaction mixture. As shown in Figure 6b, a module 2 according to the invention was located on the outer face of the side wall 80 of the container and was positioned with respect to the bioreaction mixture to be processed internally from the side wall in the interior space. Module 2 included a transmitter for electromagnetic fields. Two versions of the experiment were carried out. A first version (1) involved placing a module 2, according to the invention, in the location shown in Figure 6b within container 70. A second control version (2) involved placing a dummy device, consisting only of the module 4 casing without any components, in the same location as module 2 in version 1, within container 70. This dummy device was thus unable to transmit electromagnetic fields. Module 2 was switched on at the start of each experiment (Day 0) and placed in this location. Sampling of the antibody yields obtained from experimental versions 1 and 2 was performed on days 4, 6, 8, and 10 after switching on the module. The average results of an experimental campaign (n=6) are shown below in Table 1 and graphically in Figure 6c. QL AAΩ / 77Π7 / E / YILI Average Performance Module Day after ST.ER. Average Control configuration Module(l) Control(2) delta (1) ST.ER. Average (2) 4 18.438 14.272 0.292 3.259420802 2.294948094 6 33.171 24.610 0.348 3.30653162 3.632320054 8 38.715 32.355 0.197 4.308675478 4.632898692 ttest 10 40.608 0.003 35.137 0.156 2.662343771 2.516541155 A higher antibody yield was observed on each sampling day of experiment version 1 using module 2 compared to the control experiment version 2, with a delta of 29%, 35%, 20%, and 16% for days 4, 6, 8, and 10, respectively. A p-value of 0.003 calculated between the Module (1) experiment version and the Control (2) experiment version over the 10-day period showed that the yield increase is statistically significant, and therefore, the use of module 2 to emit electromagnetic fields is statistically and experimentally verified to increase antibody yield. In this experiment, pulsed electromagnetic fields were emitted from module 2, and the frequency of one or more of the emitted electromagnetic fields was varied during the module's operation over a predetermined period. The electromagnetic fields were emitted at three different frequencies in a random sequence: 2402 MHz, 2426 MHz, and 2480 MHz, which correspond to Bluetooth transmission protocol channels 37, 38, and 39, as illustrated in Figure 6d. The specific emission sequence of the different frequencies was random during the module's operating time period with a random delay of 0-10 milliseconds. Further experiments were conducted using a murine hybridoma (MHC) cell line and the effect of the present invention in relation thereto on IgG production. The average of the results obtained in terms of cell count, viability, and yield for each of the three runs using the invention (Version 1) are provided in the following table and graphically in Figures 8a-8c and compared with a control test run (Version 2) carried out in an identical culture but without using a module according to the invention and in the same manner and locations using the same apparatus as set out in relation to the experiments described in Figures 6a-6b. QL AAΩ / 77Π7 / E / YILI Antibody performance Experiment 1 Execution Experiment 2 Execution Experiment 3 Control (2) Active (1) Control (2) Active (1) Control (2) Active (1) Day 4 1.27 2.02 4.91 6.54 4.17 5.20 Day 6 4.28 6.46 5.00 14.85 10.69 14.88 Day 8 6.12 8.30 18.20 21.63 14.52 19.81 Day 10 7.81 12.98 19.22 26.61 19.65 22.22 delta 59.4 33.1 24.7 50.8 197.0 39.1 35.6 18.8 36.5 66.2 38.4 13.1 QL AAΩ / 77Π7 / E / YILI Table 2 The results show increased viability and slowed cell growth with the use of the module, leading to higher cell production and, consequently, increased cumulative yield. On average, across the three controlled experiments, the use of module 2 resulted in a 30% increase in IgG antibody yield compared to the conventional control. This higher production rate can be exemplified by comparing the performance of day 8 with day 10, and the use of the modules achieved a higher yield. At every point during the experiments, the measured antibody yield in the results of version 1 according to the invention arm was consistently higher than in the control version 2, indicating a higher antibody production rate. Furthermore, this increased rate was achieved with a consistently lower total cell count, suggesting that the observed productivity was a consequence of the greater sustained viability and higher productivity of the present invention. In one mode, the module can be operated continuously for the predetermined time period or, alternatively, the module is controlled to operate for a certain percentage of that predetermined time period, such as, for example, 1 hour on and 3 hours off if the total operating time period of the module is 4 hours, so that the module emits one or more electromagnetic fields for 25% of the time. Typically, the module's operation to emit one or more electromagnetic fields is pre-programmed and, in one mode, can be updated by manually entering a change 18 in the control system or by transmitting new control system data potentially wirelessly. In any case, the frequency of the electromagnetic fields and / or the selection of continuous or pulsed electromagnetic field emissions is selected, and may vary, depending on the particular type of bioreaction mixture to be processed and exposed to one or more emitted electromagnetic fields. In another embodiment of the invention, as described with respect to Figures 7a-7b, instead of the modules being located near a wall of the vessel 86, shown with dashed lines, a plurality or group of modules 2, 2', 2, 2' are placed within the interior space of the vessel and in or partially in the bioreaction mixture to be exposed to one or more electromagnetic fields. This arrangement is particularly useful when the bioreaction mixture 88 is in a condition of relatively low turbulence, i.e., little or no movement is required, such as when agitation of the bioreaction mixture is required, for example, if the process is fermentation of the bioreaction mixture.In this configuration, modules 2 are supplied with components as described above, and housing 4 is provided with a liquid-tight seal to prevent the bioreaction mixture 88 being processed from entering the module's inner cavity. Modules 2, 2', 2, and 2' are adapted so that when placed in the bioreaction mixture 88, they move to different depths within the mixture, as illustrated in Figure 7b, because the modules have different properties or intermediate buoyancy values. These different properties or values ​​are typically achieved by placing different quantities of a material of varying weight, or the same quantities of different types of material with different weights, in or on the modules.Therefore, the different intermediate buoyancy levels are used to ensure that when a group of modules is placed in the bioreaction mixture, they are at different depths within the bioreaction mixture, so that the electromagnetic fields emitted from there are emitted in different places within the bioreaction mixture to ensure that substantially uniform exposure of the bioreaction mixture to electromagnetic fields is achieved. This, in turn, means that the group user can simply place all the modules, or a selected number of modules depending on the depth of the bioreaction mixture being processed, into the bioreaction mixture and ensure that by placing the appropriate modules into the bioreaction mixture, the exposure is substantially reduced. Q ίββΩΩ / 77Ω7 / Β / YΙΛΙ uniform to the electromagnetic fields extends throughout the bioreaction mixture from the top surface to the bottom. In one embodiment, these modules include indication means 90, typically on the outer surface of the housing, to identify the modules and the buoyancy classification of the modules in the group as shown in Figure 7a. When the modules are placed on the outer surface of the container, then, although it may be preferred that the modules be placed along one or more side walls of the container to ensure substantially uniform exposure of the bioreaction mixture within the interior space to the one or more emitted electromagnetic fields, additionally or alternatively, the modules may be positioned on the base and / or top wall or lid of the interior space and / or in channels or ducts leading to or from it. In different modes, the module can be operated continuously for the predetermined time period or, alternatively, the module is controlled to operate for a certain percentage of that predetermined time period, such as, for example, 1 hour on and 3 hours off if the total operating time period of the module is 4 hours, so that the module emits the electromagnetic field or fields for 25% of the time. Typically, the module's operation to emit electromagnetic fields is pre-programmed and, in one mode, can be updated by manually entering a change in the control system or by transmitting new control system data, potentially wirelessly. In any case, the frequency of the electromagnetic fields and / or the selection of continuous or pulsed electromagnetic field emissions is selected, and may vary, depending on the particular type of bioreaction mixture that is to be processed and exposed to the emitted electromagnetic fields. One of the key advantages of the present invention is that the modules are provided to be selectively located in the positions that best suit the shape of the container and / or the bioreaction mixture to be processed at that time.

Claims

1. Apparatus for improving the performance and / or acceleration and / or quality of a bioreaction, the apparatus comprising: a container formed by a plurality of walls comprising at least one side wall and a base forming an internal space for containing a bioreaction mixture, means for causing the movement of said bioreaction mixture, and one or more modules, said module comprising a housing and a transmitter disposed within said housing, said transmitter being capable of emitting one or more electromagnetic fields, wherein the one or more modules are close to at least a portion of said walls that is made of a material that is substantially transparent to said one or more electromagnetic fields, and / or wherein the one or more modules are disposed in said internal space.

2. An apparatus according to claim 1, wherein one or more modules are arranged in said bioreaction mixture in said internal space.

3. Apparatus according to claim 1, wherein said outer casing comprises a liquid-tight coating made of a plastic material.

4. An apparatus according to any of the preceding claims, wherein said plurality of walls includes a container lid.

5. An apparatus according to claim 4, wherein said cover includes a portion made of a material that is substantially transparent to said one or more electromagnetic fields and said one or more modules are positioned close to them.

6. An apparatus according to any of the preceding claims, wherein said at least one portion made of a material substantially transparent to said one or more electromagnetic fields is made of a non-metallic material.

7. Apparatus according to any of the preceding claims, wherein said means for causing movement are electrical and / or mechanical movement means. q LRAnn / zznz / E / YiAi 8. Apparatus according to claim 7, wherein said means for causing movement comprise any, or any combination of: an agitator within the internal space of the container; an agitator external to the internal space of the container; a means for imparting vibration to the container and / or bioreaction mixture; a wave generator; an inlet material flow into the internal space and / or an outlet material flow from said internal space.

9. Apparatus according to claim 8, wherein the agitator within the internal space is an agitator, a propeller, an impeller, or a gas sprayer.

10. Apparatus according to any of the preceding claims, wherein the module(s) are located in the means for the movement of the bioreaction mixture located in the interior space.

11. Apparatus according to any of claims 1-6, wherein said means for causing movement is one or more components of the bioreaction mixture.

12. Apparatus according to any of the preceding claims, wherein said means for causing movement move said bioreaction mixture to periodically expose substantially all of said bioreaction mixture to said one or more electromagnetic fields emitted from said one or more modules and / or to retain cells in said bioreaction mixture substantially suspended in said bioreaction mixture.

13. An apparatus according to any of the preceding claims, wherein the apparatus further comprises a frame, and the container is a bag made of a plastic, elastic, and flexible material and is supported by the frame.

14. An apparatus according to claim 13, wherein one or more modules are attached to the frame.

15. An apparatus according to any of the preceding claims, wherein said electromagnetic fields emitted by the transmitter are a series of pulsed electromagnetic fields. Q LRAnn / ZZnZ / E / YIAI 16. An apparatus according to any of the preceding claims, wherein the apparatus further comprises a control means for controlling the shape of said one or more electromagnetic fields emitted from the one or more modules.

17. An apparatus according to any of the preceding claims, wherein said bioreaction mixture includes water.

18. An apparatus according to claim 17, wherein said one or more electromagnetic fields are emitted at a frequency that causes the rotation of the molecular dipole of the water contained in the bioreaction mixture.

19. An apparatus according to any of the preceding claims, wherein the control means control the emission of one or more electromagnetic fields at a frequency or frequencies in the range of 2.4 GHz to 2.5 GHz.

20. An apparatus according to any of the preceding claims, wherein said one or more electromagnetic fields are emitted at a plurality of different frequencies in a random sequence during said period of time.

21. Apparatus according to any of the preceding claims, wherein when there are two or more modules, said means of controlling the one or more electromagnetic fields emitted by them are emitted in a synchronized manner.

22. An apparatus according to any of claims 16-21, wherein the control means forms an integral part of the module, the container and / or the frame.

23. Apparatus according to any of the preceding claims, wherein the apparatus includes a support and said one or more modules are located on said support and said support can be located with the container.

24. Apparatus according to any of the preceding claims, wherein at least one module includes a rechargeable power source. Q LRAnn / ZZnZ / E / YIAI 25. Apparatus according to any of the preceding claims, wherein said transmitter provides short-range wireless communication of said one or more electromagnetic fields.

26. An apparatus according to any of the preceding claims, wherein the apparatus comprises one or more locators for locating one or more modules near or within the container.

27. An apparatus according to claim 26, wherein the one or more locators comprise one or more of a hook, a sail fastener, an adhesive patch, a threaded locator, a magnet, or any combination thereof.

28. An apparatus according to any of the preceding claims, wherein the one or more modules are arranged in the internal space and further comprise at least one flotation portion for containing the one or more modules at a depth in the bioreaction mixture when the bioreaction mixture comprises a liquid.

29. An apparatus according to claim 28, wherein there are two or more modules, at least one of which has a flotation portion of a shape selected to retain it at a different depth than another module or modules in the bioreaction mixture when the bioreaction mixture comprises a liquid.

30. An apparatus according to claim 29, wherein each of the two or more modules in a group of modules is provided with a flotation portion of a shape selected to retain said modules at different respective depths in the bioreaction mixture when the bioreaction mixture comprises a liquid.

31. An apparatus according to any of the preceding claims, wherein the container includes one or more hollow elongated members extending from at least one wall, the base, and / or the lid such that the one or more elongated members are not in fluid communication with the internal space, and wherein at least one of the one or more hollow elongated members comprises at least one portion made of a material that is transparent to electromagnetic fields. Q ίΩΩΩΩ / 77Ω7 / Β / YΙΛΙ 32. An apparatus according to any of the preceding claims, wherein the one or more modules are two or more modules, and each of the two or more modules is located at a node of another module.

33. An apparatus according to any of the preceding claims, wherein the one or more modules are two or more modules located in an array.

34. Apparatus according to claim 33, wherein said matrix extends from the base of the container upwards.

35. Apparatus according to any of claims 33-34, wherein said matrix extends along a single axis or is provided to extend along a plurality of axes.

36. Apparatus according to any of claims 33-35, wherein said modules are located in the matrix with a separation within a range of 5-10 cm between adjacent modules in the matrix.

37. Apparatus according to any of the preceding claims, wherein the one or more modules include a wireless receiver to allow the reception of signals that cause a change in at least one operating parameter of the module to be transmitted wirelessly and received by the same, and said change implemented.

38. Apparatus according to any of the preceding claims, wherein the apparatus includes a charging and / or control unit to which one or more modules, when not in use, are electrically connected and / or data connected to enable charging and / or activation control.

39. Apparatus according to any of the preceding claims, wherein said bioreaction mixture is any, or any combination of, a biofuel, cell cultures and genetically modified organisms, insulin, monoclonal antibodies, growth hormones, interferon, interleukins, blood factor VIIa, blood factor VIII, blood factor IX, erythropoietin, gonadotropin, glucagon, vaccine antigenic sequences, mammalian cell culture.

40. A kit for improving the performance and / or acceleration and / or quality of a bioreaction, the kit comprising: a container formed by a plurality of walls comprising at least one side wall and a base forming an internal space for containing a bioreaction mixture, a means for moving said bioreaction mixture, and one or more modules, said module comprising a housing and a transmitter disposed within said housing, said transmitter being capable of emitting one or more electromagnetic fields, wherein localization means are provided to allow the one or more modules to be positioned close to at least a portion of said walls made of a material that is substantially transparent to said one or more electromagnetic fields, and / or wherein the one or more modules are disposed in said internal space.

41. A kit according to claim 40, wherein the container is formed by a frame and a sheet material supported on the frame to form said internal space.

42. A method for improving the performance and / or acceleration and / or quality of a bioreaction, the method comprising the steps of: providing a container formed by a plurality of walls comprising at least one side wall and a base forming an internal space for containing a bioreaction mixture; moving said bioreaction mixture within said internal space; and providing one or more modules, said module comprising a housing and a transmitter disposed within said housing, said transmitter being capable of emitting one or more electromagnetic fields; emitting said one or more electromagnetic fields from said one or more modules into at least a portion of the internal space; and wherein the one or more modules are close to at least a portion of said walls made of a material that is substantially transparent to said one or more electromagnetic fields, and / or wherein the one or more modules are disposed within said internal space. Q LRRnn / ZZnZ / E / YIAI 43. A method according to claim 42, wherein the method comprises the step of introducing a cell culture medium into the internal space of the container and introducing one or more cells into the cell culture medium to form a bioreaction mixture.

44. A method according to any of claims 42 and 43, wherein the bioreaction mixture includes water.

45. A method according to claim 44, wherein one or more electromagnetic fields are emitted at a frequency to rotate and modulate said water molecules.

46. ​​A method according to claim 45, wherein the electric field component of the electromagnetic wave of one or more electromagnetic fields is modulated by adapting the frequency range of one or more electromagnetic fields so that the time for the hydrogen bonds to break substantially corresponds to half a cycle of the emission frequency.

47. A method according to any of claims 42-46, wherein the bioreaction mixture is moved during the application of one or more electromagnetic fields.

48. A method according to any of claims 42-47, further comprising a step of applying one or more electromagnetic fields to the cell culture medium prior to the introduction of one or more cells thereto.

49. A method according to any of claims 42-48, wherein the emission of one or more electromagnetic fields includes applying pulsed electromagnetic fields to the bioreaction mixture.

50. A method according to any of claims 42-49, wherein one or more electromagnetic fields have a frequency between 2.4 and 2.5 GHz and / or are emitted in pulses with a duration of 0.5-1.5 milliseconds (ms) and / or said pulses are separated by rest periods in the range of 40-66 ms and / or the pulses are emitted at a rate of 12-20 pulses per second. Q LRAnn / ZZnZ / E / YIAI 51. A method according to any of claims 42-50, wherein the one or more cells are yeast cells and the bioreaction is for the production of bioethanol.

52. A method according to any of claims 42-51, wherein the one or more cells are mammalian cells and the bioreaction is for the production of nucleic acids or peptides.

53. A method according to any of claims 42-52, wherein one or more cells are hybridoma cells.

54. A method according to any of claims 42-53, wherein the one or more cells are insect cells and the bioreaction is for the production of nucleic acids or peptides.

55. A module provided for emitting one or more electromagnetic fields, said module including a housing, a power source, a transmitter for emitting one or more electromagnetic fields and control means for permitting control of said one or more electromagnetic fields and wherein the one or more electromagnetic fields are controllable such that for a period of time they are emitted at; a predetermined frequency or range of predetermined frequencies and are emitted continuously or as a series of pulses with a spacing between said emission pulses and said module locatable in, or near, a bioreaction mixture such that at least part of the bioreaction mixture is within the range of the emitted electromagnetic fields.

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