Unit and method for executing a bioprocessing step

Abstract

The current invention relates to a unit for executing a bioprocessing step. In a second aspect, the present disclosure also relates to a method for executing a bioprocessing step in a bioprocessing unit. In a third aspect the present disclosure relates to the use of aforementioned unit for executing a bioprocessing step, wherein said bioprocessing step comprises the production of cells and / or the production of biomolecules such as a protein, a virus or viral particle, a cell or tissue therapy product, or a gene therapy product. In a further aspect, the present disclosure relates to a method of installing a flow path comprising a plurality of disposable tubing on aforementioned unit. In a last aspect, the disclosure relates to a method of operating a unit, wherein said unit is configured to execute a bioprocessing step in a bioreactor residing in said unit.

Description

[0001] UNIT AND METHOD FOR EXECUTING A BIOPROCESSING STEP

[0002] FIELD OF THE INVENTION

[0003] The present disclosure relates to a unit for executing a bioprocessing step. In a second aspect, the present disclosure relates to a method for executing a bioprocessing step in a bioprocessing unit. In a third aspect the present disclosure relates to the use of aforementioned unit for executing a bioprocessing step, wherein said bioprocessing step comprises the production of cells and / or the production of biomolecules such as a protein, a virus or viral particle, a cell or tissue therapy product or a gene therapy product. In a further aspect, the present disclosure relates to a method of installing a flow path comprising a plurality of disposable tubing on aforementioned unit. In a last aspect, the disclosure relates to a method of operating a unit, wherein said unit is configured to execute a bioprocessing step in a bioreactor residing in said unit.

[0004] BACKGROUND

[0005] Advanced therapy medicinal products (ATMPs) are increasingly gaining importance in the world of pharmaceuticals. Antibody-based therapies and RIMA, DNA, cell and gene therapies are deemed to provide a solution for many, hitherto not treatable diseases. In addition, due to the emergence of new viral infections such as SARS, SARS-CoV-2 and MERS, the demand for vaccine production facilities have risen.

[0006] The traditional systems for executing a bioprocessing step are often not ergonomic, easily accessible, flexible nor user-friendly. Major drawbacks related to the traditional systems known from the art is that they often cannot be operated in a small cleanroom due to the large footprint. Not only the footprint of the system itself, but also the space required for the operators to operate the system and / or the amount of operators required to operate the system often limits the implementation of the traditional systems. Not only the use of such systems in operation is difficult, but also the installation / preparation / dismantling of the disposable parts at the end / cleaning and maintenance of those traditional systems poses difficulties. Furthermore, due to their unpractical design, they often also cannot be operated along a wall. As these traditional systems often require large facilities, resulting in a negative effect on the production efficiency, the overall time for production and the total cost. Furthermore, these systems are not flexible, showing a low degree of modularity, wherein the addition of liquid reagents is severely restricted by the flow paths and their related valves and pumps, limiting for instance the range of flow rates that can be achieved. Furthermore, there is a need for user friendly systems that are modular and allow an easy installation, comprise components suitable for single-use and can operate in an automated way by a reduced amount of operators based on user-defined parameters.

[0007] The present disclosure aims to resolve at least some of the problems mentioned above. More specifically, it aims to provide a concise and modular cell and biomolecule production system, suited amongst others for the production of viral particles, proteins, and cell / tissue and gene therapy products that can be used for clinical and therapeutic purpose.

[0008] The present disclosure provides an ergonomic, highly accessible and user-friendly system which is flexible and modular. The system and method for executing a bioprocessing step according to the current disclosure allow the low-cost manufacture of cells and biomolecules, while still maintaining high quality and GMP requirements.

[0009] SUMMARY OF THE INVENTION

[0010] The present disclosure and embodiments thereof serve to provide a solution to one or more of the above-mentioned disadvantages. To this end, the present disclosure relates to a unit for executing a bioprocessing step, according to claim 1. More particular, the unit as described herein comprises: a unit for executing a bioprocessing step, wherein said unit comprises: at least one flow path, wherein the at least one flow path can be an influent or an effluent flow path, directing a liquid towards or away from a bioreactor residing in said unit; at least one pump and / or at least one valve, wherein the at least one pump and / or valve is connected to the at least one flow path; a controller, connected to said at least one pump and / or valve; a user interface, connected to the controller for allowing an operator to select the at least one flow path according to a given request sequence; wherein the controller is adapted to store said request sequence and wherein said controller is configured to initiate said request sequence, thereby controlling the liquid flow in the at least one flow path by means of activation and / or deactivation of said at least one pump and / or valve. Preferred embodiments of the unit are described in any of the claims 2 to 34.

[0011] A specific preferred embodiment relates to a disclosure according to claim 2.

[0012] In a second aspect, the present disclosure relates to a method for executing a bioprocessing step in a bioprocessing unit according to claim 35. More particular, the method as described herein comprises the steps of: installing one or more flow paths comprising a plurality of disposable tubing on said unit by means of flow path guides; installing one or more containers of choice comprising a liquid reagent on one or more supports present on said unit; connecting each of said one or more containers with a flow path of choice; providing a controller with instructions for operating the liquid flow in said flow paths, wherein said controller is adapted to store a request sequence, and wherein said controller is configured to initiate said request sequence in an operating mode, thereby controlling the liquid flow in one or more flow paths by means of activation and / or deactivation of one or more pumps and / or valves connected to said one or more flow paths.

[0013] A preferred embodiment of the method is described in claim 36.

[0014] In a third aspect the present disclosure relates to the use of aforementioned unit for executing a bioprocessing step, wherein said bioprocessing step comprises the production of cells and / or the production of biomolecules such as a protein, a virus or viral particle, a cell or tissue therapy product, or a gene therapy product.

[0015] In a further aspect, the present disclosure relates to a method of installing a flow path comprising a plurality of disposable tubings on the aforementioned unit.

[0016] In a last aspect, the disclosure relates to a method of operating a unit, wherein said unit is configured to execute a bioprocessing step in a bioreactor residing in said unit. More particular, the method as described herein comprises selecting one or more flow paths via a user interface of said unit according to a given request sequence, wherein said flow path can be an influent or an effluent flow path, directing a liquid towards or away from said bioreactor residing in said unit, wherein said user interface is connected to a controller, said controller stores said request sequence, which is initiated by said controller in the operating mode of said unit, thereby controlling the liquid flow in a given flow path by means of activation and / or deactivation of one or more pumps and / or valves of said unit. DESCRIPTION OF FIGURES

[0017] Figures 1-4 show schematic representations (left front side, left side, right front side and back view, respectively) of the unit according to an embodiment of the present disclosure.

[0018] Figure 5 shows a general overview of a flow diagram of the unit according to an embodiment of the present disclosure.

[0019] Figures 6-24 show flow diagrams of various request sequences according to an embodiment of the present disclosure.

[0020] Figures 25-27 show tags (A1-A4 and B1-B4) assigned to one or more tubing supports to facilitate the installation of the tubing according to an embodiment of the present disclosure. The tags can be engraved on a panel of the unit (Figure 25), can be attached to the unit by means of an adhesive (Figure 26) or can comprises a LED system comprising LED lights (Figure 27) controlled by the controller of the unit according to an embodiment of the present disclosure.

[0021] Figure 28 depicts an assembly enabling aseptic liquid sampling for instance from the bioreactor or from the harvest tank according to an embodiment of the present disclosure.

[0022] Figure 29 depicts a holder designed to support the liquid sampling assembly and facilitate drawing of samples according to an embodiment of the present disclosure.

[0023] Figures 30-32 depict dimensions of a unit according to an embodiment of the present disclosure and shows tags engraved on the unit according to an embodiment of the present disclosure to facilitate installation of tubing and containers.

[0024] Figures 33-34 show schematic representations (left front side, right front side, respectively) of the unit according to an embodiment of the present disclosure also including suspended transfer bags (which are used as containers connected to specific flow paths).

[0025] Figure 35 shows a side panel of the unit including manifolds according to an embodiment of the present disclosure. Figure 36 shows an assembly for directing a liquid towards the bioreactor of the unit ("BIO IN") according to an embodiment of the invention.

[0026] Figure 37 shows an assembly for directing a liquid away from the bioreactor of the unit ("BIO OUT") according to an embodiment of the invention.

[0027] Figure 38 shows an assembly for a foam trap of the unit according to an embodiment of the invention.

[0028] Figure 39 shows an assembly for small additions to the bioreactor according to an embodiment of the invention.

[0029] Figure 40 shows an assembly for large additions to the bioreactor according to an embodiment of the invention.

[0030] Figure 41 shows a base assembly of the unit according to an embodiment of the invention.

[0031] Figure 42 shows a base bottle assembly of the unit according to an embodiment of the invention.

[0032] Figure 43 shows a transfer bag assembly (5 liters) of the unit according to an embodiment of the invention.

[0033] Figure 44 shows a transfer bag assembly (10 liters) of the unit according to an embodiment of the invention.

[0034] DETAILED DESCRIPTION OF THE INVENTION

[0035] The present disclosure relates to a unit for executing a bioprocessing step, a method for executing a bioprocessing step in a bioprocessing unit, use of aforementioned unit for executing a bioprocessing step, a method of installing a flow path comprising a plurality of disposable tubings on said aforementioned unit and a method of operating a unit.

[0036] Unless otherwise defined, all terms used in disclosing the disclosure, including technical and scientific terms, have the meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. By means of further guidance, term definitions are included to better appreciate the teaching of the present disclosure.

[0037] As used herein, the following terms have the following meanings:

[0038] "A", "an", and "the" as used herein refers to both singular and plural referents unless the context clearly dictates otherwise. By way of example, "a compartment" refers to one or more than one compartment.

[0039] "About" as used herein refers to a measurable value such as a parameter, an amount, a temporal duration, and the like, is meant to encompass variations of + / - 20% or less, preferably + / -10% or less, more preferably + / -5% or less, even more preferably + / -1% or less, and still more preferably + / -0.1% or less of and from the specified value, in so far such variations are appropriate to perform in the disclosed invention. However, it is to be understood that the value to which the modifier "about" refers is itself also specifically disclosed.

[0040] "Comprise", "comprising", and "comprises" and "comprised of" as used herein are synonymous with "include", "including", "includes" or "contain", "containing", "contains" and are inclusive or open-ended terms that specifies the presence of what follows e.g. component and do not exclude or preclude the presence of additional, non-recited components, features, element, members, steps, known in the art or disclosed therein.

[0041] Furthermore, the terms first, second, third and the like in the description and in the claims, are used for distinguishing between similar elements and not necessarily for describing a sequential or chronological order, unless specified. It is to be understood that the terms so used are interchangeable under appropriate circumstances and that the embodiments of the invention described herein are capable of operation in other sequences than described or illustrated herein.

[0042] The recitation of numerical ranges by endpoints includes all numbers and fractions subsumed within that range, as well as the recited endpoints.

[0043] The expression "% by weight", "weight percent", "%wt" or "wt%", here and throughout the description unless otherwise defined, refers to the relative weight of the respective component based on the overall weight of the formulation. Whereas the terms "one or more" or "at least one", such as one or more or at least one member(s) of a group of members, is clear per se, by means of further exemplification, the term encompasses inter alia a reference to any one of said members, or to any two or more of said members, such as, e.g., any >3, >4, >5, >6 or >7 etc. of said members, and up to all said members.

[0044] Unless otherwise defined, all terms used in disclosing the invention, including technical and scientific terms, have the meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. By means of further guidance, definitions for the terms used in the description are included to better appreciate the teaching of the present invention. The terms or definitions used herein are provided solely to aid in the understanding of the invention.

[0045] Reference throughout this specification to "one embodiment" or "an embodiment" means that a particular feature, structure or characteristic described in connection with the embodiment is included in at least one embodiment of the present invention. Furthermore, the particular features, structures or characteristics may be combined in any suitable manner, as would be apparent to a person skilled in the art from this disclosure, in one or more embodiments. Furthermore, while some embodiments described herein include some but not other features included in other embodiments, combinations of features of different embodiments are meant to be within the scope of the invention, and form different embodiments, as would be understood by those in the art. For example, in the following claims, any of the claimed embodiments can be used in any combination.

[0046] "Plurality" as used herein refers to a number greater than one.

[0047] Perfusion is a process step where cells in a bioreactor are continuously fed with fresh medium and at the same time an equal amount of spent medium is removed, which enables high density cell growth. The perfusion rate can vary during the process depending on the type of cell line used, the polypeptide product produced by those cells, the specific cell culture medium employed and the cell growth system (for instance a fixed bed) used. An important aspect in the removal of the spent medium is also to remove the (possible toxic) metabolic side products from the cell culture. These side products may have a negative effect on cell viability, and further may impair the productivity of producer or host cells. Furthermore, it also allows to harvest a product of interest in case the product is secreted by the cells. Perfusion hence could also be used to harvest a secreted / released product by the cells.

[0048] Options for perfusion include batch perfusion and fed-batch perfusion (where feeding of fresh medium is performed but no removal of spent medium is performed) or a mixed process combining different feeding strategies.

[0049] Different feeding strategies (operating modes) of the bioreactor and combinations of these strategies can be implemented in the current disclosure, including:

[0050] • batch mode (just the bioreactor without feed-in-feed-out, no addition or removal of medium)

[0051] • fed batch (bioreactor with punctual feed-in and potentially also feed-out - several additions possible and potentially several lines for feed-in and feed- out)

[0052] • perfusion (continuous feed-in and feed-out but at potentially variable flow rates)

[0053] • recirculated batch (perfusion with feed-in and feed-out connected to the same container)

[0054] The bioprocessing step of the current disclosure comprises the production of cells and / or the production of biomolecules.

[0055] "Production of cells" can for instance relate to the use of "seed trains" during cell culture. In upstream bioprocessing, a seed train scales a small volume of cell culture to a larger volume for instance to inoculate intermediate bioreactors, and eventually the production bioreactor. The purpose of a seed train is the generation of an adequate number of cells for the inoculation of a production bioreactor. The cells are usually run through many cultivation systems which become larger with each passage.

[0056] "Production of cells" can for instance also relate to cultivating or growing cell masses for tissue engineering, cultivating meat or seafood products, tissue or leather etc. Lab-grown or cultured meat belongs to the emerging field of cellular agriculture and represents a promising technology for delivering products that have so far been produced through livestock. This technological innovation aims to offer a possibility of reducing the negative effects of conventional meat production techniques on humans, livestock, and the environment. "Production of cells" can also for instance relate to a process involving the production of cells for use in cell therapy and typically refers to the cultivation, expansion, and processing of therapeutic cells intended for medical treatment. Examples include T- cells for immunotherapy (e.g., CAR-T cells), stem cells (e.g., mesenchymal or hematopoietic stem cells), or other genetically modified cells.

[0057] "Production of cells" can also for instance relate to a process involving cell biomass production for cell banking. "Cell banking" as used herein refers to establishing master or working cell banks that are preserved and cataloged for future use, whether for R&D, vaccine production, or future therapeutic manufacturing.

[0058] Large volumes of cells (e.g., CHO cells, HEK293, yeast, algae, etc.) can for instance be produced for downstream protein expression (biologies), extracting cellular components (lipids, enzymes) or metabolic engineering applications (e.g., biosynthesis of pharmaceuticals or chemicals).

[0059] Advanced therapy medicinal products (ATMPs) are medicines that are based on genes, tissues or cells. They offer groundbreaking new opportunities for the treatment of disease and injury. ATMPs can be classified into three main types:

[0060] - gene therapy medicines: these contain genes that lead to a therapeutic, prophylactic or diagnostic effect. They work by inserting 'recombinant' genes into the body, usually to treat a variety of diseases, including genetic disorders, cancer or long-term diseases.

[0061] - somatic-cell therapy medicines: these contain cells or tissues that have been manipulated to change their biological characteristics or cells or tissues not intended to be used for the same essential functions in the body. They can be used to cure, diagnose or prevent diseases;

[0062] - tissue-engineered medicines: these contain cells or tissues that have been modified so they can be used to repair, regenerate or replace human tissue.

[0063] In addition, some ATMPs may contain one or more medical devices as an integral part of the medicine, which are referred to as combined ATMPs. An example of this is cells embedded in a biodegradable matrix or scaffold.

[0064] "Biomolecule" refers to any biological material of interest that is produced in a bioreactor. Biomolecules include, for example, viruses, virus-like particles, viral products, extracellular vesicles, cell or tissue therapy products, gene therapy products, viral vectors, DNA, RNA (for instance mRNA), proteins such as antibodies, carbohydrates, lipids, nucleic acids, metabolites and peptides. "Gene therapy product" refers to a therapeutic product comprising nucleic acids to treat or prevent a disease or disorder, such as a genetic disease or disorder.

[0065] "Gene therapy product" hence refers to a medicinal product that contains or consists of recombinant nucleic acids (such as DNA or RIMA), delivered by means such as viral vectors or non-viral delivery systems, and is administered to regulate, repair, replace, add, or delete a genetic sequence in human cells, with the intent to treat, prevent, or cure a disease or disorder, including genetic conditions.

[0066] "Cell therapy product" or "Cellular therapy product" includes cellular immunotherapies, cancer vaccines, and other types of both autologous and allogeneic cells for certain therapeutic indications, including hematopoietic stem cells and adult and embryonic stem cells.

[0067] "Stem cells" are naturally occurring cells in the body that have the ability to divide and produce a range of different cell types. Stem cells are important in the growth and development of the body, as well as in repair after injury. Stems cells are categorised as ATMPs when these cells undergo substantial manipulation or are used for a different essential function. They can be somatic-cell therapy products or tissue-engineered products, depending on how the medicine works in the body.

[0068] A "tissue therapy product" or a "tissue-engineered product" relates to a medicine containing engineered cells or tissues, which is intended to regenerate, repair or replace a human tissue.

[0069] "Viral gene therapy product" refers to a viral product where a part of the genetic material of the virus is substituted with therapeutic nucleic acids and where the virus is implemented to introduce the therapeutic nucleic acids into the cells of the patient. A number of viruses have been used for human gene therapy, including retroviruses, adenoviruses, herpes simplex, vaccinia, and adeno-associated virus.

[0070] "Antibody" refers to any immunoglobulin molecule, antigen-binding immunoglobulin fragment or immunoglobulin fusion protein, monoclonal or polyclonal, derived from human or other animal cell lines, including natural or genetically modified forms such as humanized, human, chimeric, synthetic, recombinant, hybrid, mutated, grafted, and in vitro generated antibodies. Commonly known natural immunoglobulin antibodies include IgA (dimeric), IgG, IgE, IgG and IgM (pentameric). "Virus" or "virion" refers to an ultramicroscopic (roughly 20 to 300 nm in diameter or larger sometimes), infectious agent that replicates only within the cells of living hosts, mainly bacteria, plants, and animals: composed of an RIMA or DNA core, a protein coat, and, in more complex types, a surrounding envelope.

[0071] "Bioreactor" refers to any device or system that supports a biologically active environment, for example for cultivation of cells or organisms for production of a biological product or biomolecule. This would include cell stacks, roller bottles, shakes, flasks, stirred tank suspension bioreactors, structured or unstructured fixed- bed bioreactors, high cell density structured or unstructured fixed-bed bioreactors, batch reactors, perfusion bioreactors, etc. In an embodiment, the bioreactor is developed for single-use and comprises disposable pre-fitted manifolds, for instance pre-fitted manifolds for a top and bottom liquid bioreactor drain, a liquid sample line, a bubble or foam trap, a base addition line, etc.

[0072] As used herein, "viral infection" refers to the entry of a virus into a cell and the subsequent replication of the virus in the cell.

[0073] "Cell infection" as used herein relates to natural infection of cells by viruses, thereby attaching to receptors and injecting their genetic material. This is commonly used to study viruses and produce viral vectors for gene therapy.

[0074] "Transfection" as used herein relates to introducing foreign genetic material into cells using non-viral methods like chemicals, electricity, or microinjection. Transfection is widely used to study gene function, manipulate gene expression, and produce recombinant proteins.

[0075] "Inoculation" as used herein refers to the process of introducing a specific quantity of live cells into a bioreactor to initiate or maintain a biological reaction. This step is critical in bioprocessing, as it sets the stage for cell growth, production of desired metabolites, or other biological activities. The inoculated cells proliferate and perform their intended functions within the controlled environment of the bioreactor. The inoculum is typically prepared from a cell culture that has been cultivated to a specific growth phase (usually the exponential phase) to ensure optimal viability and activity. In an embodiment, the inoculum is collected from a smaller bioreactor and transferred to the bioreactor residing in the unit of the current disclosure by means of a transfer bottle. "Cell culture harvest", "culture harvest" and "harvest" are used as synonyms and refer to any portion of liquid originating from the bioreactor comprising cells and / or biomolecules produced by said cells. It thus relates to any product or intermediate product obtained from culturing cells in a bioreactor. The cultured cells or the grown cells also are referred to as host cells.

[0076] As used herein, "docking" means to make a stable connection between two elements, whereby the elements can for instance comprise either a receiving portion or a connecting portion. In this disclosure, docking can for instance occur between the bioreactor and the bioreactor docking station of the unit.

[0077] Detailed description

[0078] Bioprocessing steps for cell biomass expansion and / or production of biomolecules such as a protein, a virus or viral particle, a cell or tissue therapy product or a gene therapy product require a unit which is able to perform a multitude of steps. Preferably the unit supports multiple steps during a cell culture and / or a biomolecule production process, including for instance inoculation of cells, transfection, transduction, cell growth and production of biomolecules. During these steps, additional actions need to be performed, such as liquid sampling from the bioreactor, complete or partial draining of the bioreactor, complete or partial filling of the bioreactor and addition of various solutions to the bioreactor. These solutions may comprise liquid reagents that can for instance include cell culture media, cell nutrients, a transfection or infection mix, various buffers, enzyme solutions or other additives, which need to be added to the bioreactor under the correct conditions. For instance, in certain embodiments, for long duration culture, it may be advantageous to replace part of the culture medium with fresh medium or to carry out an addition of nutriment or reagent.

[0079] It is of great importance that the correct amount of the desired liquid reagent is added at the desired time point and at a correct flow rate to the bioreactor.

[0080] As such, in a first aspect, the invention relates to a unit for executing a bioprocessing step, wherein said unit comprises: at least one flow path, wherein the at least one flow path can be an influent or an effluent flow path, directing a liquid towards or away from a bioreactor residing in said unit; at least one pump and / or at least one valve, wherein the at least one pump and / or valve is connected to the at least one flow path; a controller, connected to said at least one pump and / or valve; a user interface, connected to the controller for allowing an operator to select the at least one flow path according to a given request sequence; wherein the controller is adapted to store said request sequence and wherein said controller is configured to initiate said request sequence, thereby controlling the liquid flow in the at least one flow path by means of activation and / or deactivation of said at least one pump and / or valve.

[0081] In contrast to traditional systems, the unit of the current disclosure has a limited number of flow paths which can be reused for the consecutive addition / removal of different components (for instance liquid reagents to be added or waste to be removed). The unit of the current invention is hence able to support multiple steps during a cell culture and / or a biomolecule production process, including inoculation of cells, cell growth and production of biomolecules using said limited number of flow paths. In a preferred embodiment, the flow paths comprise a plurality of disposable tubing. In an embodiment, the same tubing can be used for the consecutive addition / removal of multiple components. A limited number of flow paths can be used during operation, because the unit of the current invention enables draining or emptying of the tubing in order to reuse them for a different purpose. In some embodiments, rinsing or priming the tubing for another use may be necessary. In an alternative embodiment, the tubing is disposable and can be replaced when a different component is to be added using the same flow path. As described above, said flow path can comprise multiple sections of tubing, wherein two consecutive sections of tubing are separated the one from the other by positioning of various components in between said sections of tubing (e.g. a container, a valve, an aseptic connector, a sensor, a split tubing assembly, etc.). In another embodiment, said flow path can consist of one uninterrupted single piece of tubing directly connecting a container of choice with the bioreactor.

[0082] Components that can be added for instance include: medium to the bioreactor (for instance during perfusion mode), one or more buffers (lysis buffer, washing buffer, rinsing buffer, ...), reagents additions during the process, for instance for changing the pH, conductivity, osmolarity, endonucleases, stabilizer, media boost, or for infection / transfection, etc. Components that can be removed for instance include: medium from the bioreactor (for instance during perfusion mode), medium samples from the bioreactor, a cell or biomolecule harvest, etc.

[0083] As each flow path is connected to a specific pump or valve, the operator can control the flow rate and the duration and timing of the addition / removal, thereby taking into account the shear sensitivity and the volume of the liquid reagent to be added / removed.

[0084] As such, the invention relates to a unit for executing a bioprocessing step. In a preferred embodiment, said unit is a cabinet, said cabinet comprising one or more side panels, one or more front panels and one or more back panels (see figures 1-4 for a preferred embodiment of the lay-out of the unit). In an embodiment, said unit comprises an internal storage unit having one or more compartments. In an embodiment, the one or more compartments can be accessed by doors provided in one or more front panels, side panels and / or back panels of the unit. In an embodiment, the one or more compartments can be accessed by doors provided in one or more front panels (see figures 1 and 3). In an embodiment, the one or more compartments can be accessed by doors provided in one or more back panels (see figure 4). In an embodiment, one or more of said doors may be sliding doors and / or one or more of said doors may be removable. Said one or more back panels are by preference a vertical metal sheet. Said one or more back panels can however be made from any suitable material known in the art, such as metal, plastic or acrylic (plexiglass) back panels. In an embodiment, modular back sheet templates, different back sheets with specific hardware, can be designed, depending on the processes to be executed in the system, allowing interchangeability and easy development for future customer need or requirement. The back sheets can be adapted and assembled together to fit different process needs. Furthermore, the combination and position of these back sheets can be adjusted onsite at any point in order to facilitate multi-product manufacture. In an embodiment, in the back of this sheet all technical components are installed like motors, network cables, power supply, etc. This design allows modularity, easy maintenance, increased accessibility, a decreased amount of necessary components and increases safety. In an embodiment, one or more of said back sheets are interchangeable. In an embodiment, one or more of said components installed on said sheets are interchangeable.

[0085] Major drawbacks related to the traditional systems know from the art is that they often cannot be operated in a small cleanroom due to the large footprint. Not only the footprint of the system itself, but also the space required for the operators to operate the system and / or the amount of operators required to operate the system often limits the implementation of the traditional systems. Not only the use of such systems in operation is difficult, but also the installation / preparation / dismantling of the disposable parts at the end / cleaning and maintenance of those traditional systems poses difficulties. Not only the system itself, but also the containers comprising a liquid reagent which are connected to a chosen flow path often require a lot of space in the traditional systems. Furthermore, due to their unpractical design, often the traditional systems also cannot be operated along a wall. As these traditional systems often require large facilities, resulting in a negative effect on the production efficiency, the overall time for production and the total cost.

[0086] In an embodiment, the unit is adapted to be juxtaposed against a second unit or wall. In an embodiment, the back panel of the unit is adapted to be positioned against a wall (see figures 1-4).

[0087] Production of cells and / or the production of biomolecules requires bioprocessing steps which necessitate the flow of liquid towards and from the bioreactor. Traditional systems are often not flexible, showing a low degree of modularity, wherein the addition of liquid reagents is severely restricted by the flow paths and their related valves and pumps, limiting for instance the range of flow rates that can be achieved. See Table 1 below for a comparison of various pump types and gravity addition.

[0088] Table 1

[0089] The unit of the invention provides a plurality of flow paths. As described above, said flow paths can be influent or effluent flow paths, directing a liquid towards or away from a bioreactor residing in said unit, respectively. In an embodiment, said flow path can be an influent flow path at a first time point and an effluent flow path at a second time point, or vice versa. As such, the flow path can be used to direct a liquid towards a bioreactor residing in said unit at a certain time point and can be used to direct a liquid away from a bioreactor residing in said unit at another time point. In an embodiment, flow paths are not limited to directing a liquid towards or away from a bioreactor residing in said unit, but are used to move liquid in and out of other points in the unit.

[0090] In a preferred embodiment, said flow path is realized by tubing, preferably disposable tubing. Said disposable tubing can have any suitable dimensions. In an embodiment, the internal diameter (ID) of one or more disposable tubings used in the system is comprised between 1-20 mm, such as between 3 and 15 mm. In an embodiment, the outside diameter (OD) of one or more disposable tubings used in the system is comprised between 1-25 mm, such as between 5 and 20 mm. In an embodiment, one or more disposable tubings used in the system have an internal diameter (ID) of 1 / 8" and an outside diameter (OD) of 1 / 4". In an embodiment, said ID is 1 / 4" and said OD is 7 / 16". In an embodiment, said ID is 3 / 16" and said OD is 3 / 8". In an embodiment, said ID is 3 / 8" and said OD is 5 / 8". In an embodiment, said ID is 1 / 2" and said OD is 3 / 4". It will be clear to the skilled person that these dimensions also can apply to non-disposable tubing and are not limited to disposable tubing.

[0091] Said tubing can be made from any material known from the art. The tubing may be disposable, sterile, and provided as a ready-to-use component. Suitable tubing materials may further include, without limitation, C-Flex®, PharMed®, neoprene, silicone or platinum-coated silicone or weldable thermoplastic elastomer (TPE) tubings. It is understood that the invention is not limited to these examples, and any biocompatible, process-compatible tubing material known in the field may be used.

[0092] Said disposable tubing can be made from any material known from the art, such as Polyvinyl chloride (PVC) and silicone. In an embodiment, said tubing comprises platinum-cured silicone tubing.

[0093] In an embodiment, at least part of said flow paths comprise one or more sensors and / or indicators for measuring and indicating parameters such as flow rate, pressure and / or temperature. In an embodiment, at least one of said flow paths comprises one or more filters.

[0094] In an embodiment, at least part of said influent flow paths are positioned such that they provide a slope of between 1.0 and 3.0 degrees from a horizontal plane.

[0095] The slope can be defined as the angle a between a straight line from one end of a flow path to an opposite end of the flow path and the horizontal plane h. Suitably, the slope of one or more flow paths can be at least 1.0 degrees, such as between 1.0 and 20.0 degrees, between 1.0 and 10.0 degrees, between 1.0-3.0 degrees, such as 1.0, 1.5, 2.0, 2.5, 3.0 or any other value between 1.0 and 3.0 degrees from the horizontal plane. The slope allows efficient draining of the liquid reagents added to the flow paths towards or from the bioreactor. This also minimizes the amount of liquid remaining in the flow path and eventually discarded after use when using a disposable flow path, facilitating disposal of the flow path and reducing the amount of potentially biohazardous material to be handled as waste. Further, removal of air from the system is also facilitated by the slope.

[0096] The slope is particularly important when a flow path with lengths of tubing connected by hose barb couplings is used, as the lower inner diameter of the hose barb couplings causes stagnating pools and can trap air bubbles. Hose barb couplings are desirable for braided tubing and other types of tubing which is not amenable to connection by welding or molding. However, in a preferred embodiment, at least part, more preferably all, of said influent flow paths are uninterrupted paths, meaning that they are not interrupted by a connection, such as those formed by welding, molding or hose barb couplings. This will prevent the formation of stagnating pools and trapping of air bubbles. In an embodiment, the unit of the invention comprises flow path guides present on one or more panels of said unit. Said flow path guides can be for instance visually and / or tactilely distinguishable lines for the visualization of said flow paths. Such visualization of the flow paths can be used for the correct placement of tubes in alignment with the flow path. Said guides can be for instance black lines, colored lines or relief lines (for instance obtained by painting, printing, laser engraving and / or chemical etching) along which the disposable tubes of a flow path can be arranged. In an embodiment, said guides are laser-engraved and / or chemically etched on one or more panels of said unit. In an embodiment, the unit of the invention comprises markings along the flow path to facilitate the installation of the flow path. In an embodiment, said marking comprises a tag. Said tag can for instance be painted or printed on the panels of the unit, engraved in the panels of the unit or be attached to the unit by means of an adhesive (the tag can be for instance a sticker). In another embodiment, said marking comprises a LED system comprising LED lights or any other visual indicator. In a further embodiment, the guides comprise LED lights that are correlated to a request sequence in the controller and during a setup portion of the instrument, the LED light will light up and guide the installation of the manifold. Other visual indicators for instance include color-coded lights, flashing signals, or illuminated symbols, to assist the operator in the correct assembly or positioning. In an embodiment, these visual indicators include but are not limited to flashing lights, which intermittently activate to attract attention or signal specific states or setup steps; color-coded illumination, whereby different colors such as red, green, blue, or amber are used to convey status, position, readiness, or error conditions; and blinking patterns, where predefined sequences of light pulses correspond to particular instructions or alerts. In an embodiment, the visual indicator may take the form of an alphanumeric display, such as an LED or LCD screen configured to present letters, numbers, or symbols that indicate status messages or installation instructions. Alternatively, the visual indicator may involve symbolic projections or icons, including lights that project or illuminate shapes, arrows, or pictograms onto a surface to guide the user.

[0097] In an embodiment, said flow path guides and / or markings are used for the correct placement of tubes in alignment with the flow path. In an embodiment, said flow path guides and / or markings are used for the correct placement of pumps and / or valves in alignment with the flow path. In an embodiment, said flow path guides and / or markings are used for the correct placement of one or more containers comprising a liquid reagent connected to a chosen flow path. In an embodiment, said flow path guides and / or markings are used to indicate the flow direction of the flow path. In an embodiment, said flow path guides and / or markings are used to differentiate between punctual and continuous use of tubes, pumps, valves and / or containers. In an embodiment, said flow path guides and / or markings are used to differentiate between flow paths comprising liquid and gas lines. Said flow path guides and / or markings can be positioned on any of the panels of the unit, both on the side facing the interior of the unit and on the exterior facing side of the panels of the unit.

[0098] In an embodiment, said guides are adapted to receive said flow path, said guides than function as a support for the tubing and / or the containers. In an embodiment, said guides are clamps allowing to secure the flow path to one or more panels of the unit. As such, the guides may comprise means for restraining a a disposable flow path (such as tubing) with the desired slope, e.g. pegs, ledges and / or recesses adapted to receive the disposable flow path. The disposable tubes of the flow path can suitably be aligned with the guides, and in case the guides comprise restraining means this can minimize any slack in flexible flow path tubes. During use, single use systems for treatment of bioprocess liquids require frequent installation and removal of the flow path tubing. This is greatly facilitated by the presence of guides.

[0099] In an embodiment, a tag is assigned to one or more tubing supports to facilitate the installation of the tubing. As discussed above, said tags can for instance be engraved on a panel of the unit (figure 25), can be attached to the unit by means of an adhesive (figure 26) or can comprises a LED system comprising LED lights (figure 27) controlled by the controller of the unit.

[0100] In an embodiment, the tubing is replicated or shown on the user interface and progress of the tubing installation is tracked through manual entry or sensor integration at specific locations along the guide lines, allowing for identification or errors in the installation process.

[0101] In an embodiment, the user interface allows to track the liquid flow inside the unit.

[0102] Said guides adapted to receive said flow paths are preferably positioned on one or more of said panels in such a way to prevent crossing of one or more of said flow paths and / or increase visibility of the liquid flow in one or more flow paths. In a preferred embodiment, the different flow paths of the unit cross each other as little as possible, as this would not only make the installation of the flow path more difficult, but would also possibly decrease the visibility of the liquid flowing in said flow path when tubing is used that allows to make the liquid flow visible.

[0103] In an embodiment, the majority of the influent flow paths is positioned at the front panel and the side panels of the unit and is visible to the operator. Preferably at least 50%, more preferably at least 60%, at least 70% or at least 80% of the total influent flow path length, meaning the sum of length of the different influent flow paths present in the unit, are positioned at the front panel and the side panels of the unit. In an embodiment, the guides on the front panel of the unit are positioned as such to prevent at least 50%, more preferably at least 60% of the total length of the influent flow paths positioned on said front panel from crossing each other.

[0104] In an embodiment, the guides on the front panel of the unit are positioned as such to provide at least 50%, more preferably at least 60% of the total length of the influent flow path positioned on said front panel with a slope of at least 1.0 degrees, such as between 1.0 and 20.0 degrees, between 1.0 and 10.0 degrees, between 1.0 and 3.0 degrees from a horizontal plane.

[0105] The required bioprocessing steps should be performed at the right time for the right amount of time with a desired flow rate. These steps could follow a given request sequence. In an embodiment, said request sequence is preprogrammed. In an embodiment, said request sequence can be chosen by an operator and instructed to the controller of the bioprocessing unit by means of a user interface.

[0106] In an embodiment, an operator can select one or more flow paths according to a given request sequence in a start-up mode (hence before the unit is in operation and no bioprocessing step has been performed).

[0107] In an embodiment, an operator can select one or more flow paths according to a given request sequence in an operating mode (when the unit is in operation and one or more bioprocessing steps have been performed).

[0108] Said request sequence can be instructed to and stored by the controller. Said controller is configured to initiate said request sequence. Instructions to initiate said request sequence can be preprogrammed and stored by the controller or can be manually instructed by the operator to the controller. In an embodiment, said the unit is remotely operated or controlled, for instance by means of a remote computer, a remote server, or a network. In an embodiment, said request sequence can be preprogrammed (for instance by the distributor of the unit). In an embodiment, said request sequence can be programmed by an operator, wherein the operator can choose and program the different steps of a request sequence. In an embodiment, the various steps are preprogrammed and can simply be selected by the operator from a list of possible steps, thereby building the desired request sequence. In an embodiment, the request sequence can be manually paused or prematurely terminated. In an embodiment, additional steps can be added to the request sequence during operation.

[0109] Such a request sequence can for instance include, but is not limited to, one or more of the following steps:

[0110] - filling of the bioreactor with growth medium;

[0111] - equilibration of the medium inside the bioreactor to obtain a stable pH and / or dissolved oxygen (DO) concentration; liquid sampling of the medium in the bioreactor; inoculation of cells in the bioreactor in batch mode, including the removal of a volume equivalent to the inoculum volume and subsequent filling of the bioreactor with an inoculum comprising cells; inoculation of the cells in recirculation mode; cell growth in batch mode allowing cells to attach to the growth matrix inside the bioreactor; cell growth in fed-batch liquid sampling during batch mode to verify the attachment of the cells to the growth matrix and check the mortality rate of the cells which remained in suspension; cell growth in recirculation mode during which media is recirculated; cell growth in perfusion mode during which fresh medium is continuously added and used medium is removed; cell growth in fed batch mode during which partial or complete emptying of the medium inside the bioreactor is followed by addition of an equal volume of fresh medium; one or more washing steps;

[0112] - transfection or infection of the cells inside the bioreactor in batch mode;

[0113] - transfection or infection of the cells inside the bioreactor in recirculation mode; production of the desired biomolecule in perfusion mode and / or batch mode, and / or fed-batch and / or in recirculation mode; harvesting of the desired biomolecule if the latter is secreted or released by the cells; lysis of the cells if the desired biomolecule is not secreted by the cells or only partially secreted using lysis buffers; drainage of fluids from the bioreactor; cell harvest using enzymes; one or more rinsing steps.

[0114] - Addition of one or more buffers and / or reagents additions during the process, for instance for changing the pH, conductivity, osmolarity, adding lysis buffer, endonucleases, stabilizer, media boost, or for infection / transfection, etc.

[0115] The unit of the current invention is optimally configured to perform one or more of these steps, having a plurality of flow paths, wherein each of said flow paths can be an influent or an effluent flow path, directing a liquid towards or away from a bioreactor residing in said unit and having one or more pumps and / or one or more valves, wherein each of said one or more pumps and / or valves is connected to a given flow path. In an embodiment, at least a portion of the one or more flow paths is reused for another request sequence or to repeat the request sequence. In an embodiment, at least a portion of the one or more flow paths is reused for at least one other or different material. In an embodiment, at least a portion of the one or more flow paths is reused for another step of a request sequence or to repeat a step in the request sequence.

[0116] The request sequence specifies the steps to be performed by controlling the liquid flow in a given flow path by means of activation and / or deactivation of said one or more pumps and / or valves. As described above, in an embodiment, said request sequence can be preprogrammed and the operator can select such a preprogrammed request sequences. In an embodiment, the operator can create his or her own request sequence.

[0117] The liquid flow can be controlled by one or more pumps or by gravity addition.

[0118] Shear sensitive products should preferably by added to the unit by means of a system that minimizes shear on the product. Gravity addition minimizes shear and further also allows to add small volumes. Pumps can also be used to control the liquid flow.

[0119] In an embodiment, said pumps are chosen from low shear pumps or peristaltic pumps. In an embodiment, the unit comprises one or more reversible pumps, such as a positive displacement pump.

[0120] In an embodiment, said pumps are chosen from low flow rate pumps and high flow rate pumps. By selecting a given flow path connected to a certain pump, the operator can choose the flow rate at which the liquid reagent is added or removed from the bioreactor.

[0121] Generally speaking, fluids fall into one of two categories: Newtonian and NonNewtonian. Among other things, a Newtonian fluid holds its viscosity regardless of shear rate. Conversely, the viscosity of non-Newtonian liquids changes depending on shear rate. Shear is defined as relative motion between adjacent layers of a moving fluid. When one layer of a fluid moves adjacently to another layer of fluid, it can begin to exhibit the deleterious effects of shear. Shear rate is defined as the measure of the extent or rate of relative motion between adjacent layers of a moving fluid.

[0122] Shear sensitive products (such as liquids comprising viruses, viral vectors produced and transfection mix or sensitive cells and lysates containing valuable proteins), should be pumped with a pump conferring a low shear, such as a diaphragm pump. In an embodiment, a Quattroflow pump, a quaternary diaphragm pump that uses an eccentric disc to move fluid through the pump chamber is used to pump shear sensitive liquids. Other examples of low shear pumps include positive displacement pumps (e.g. piston pumps), progressive cavity pumps, eccentric disc pumps, sine pumps, twin screw pumps, low shear centrifuge pumps, low shear peristaltic pumps with specific roll bearings, etc. Diaphragm pumps move liquids through suction created by a vibrating diaphragm, while peristaltic hose pumps move liquids along by squeezing liquid-filled hose with a one-way "milking" action.

[0123] Diaphragm pumps are generally more complex, and have more valves, so they often require more technical knowledge initially. Peristaltic pumps are simpler, and work more easily under difficult conditions, yet over time the hose requires more attention. On the downside, diaphragm pumps work best when the liquid is clean and free of particles or gases and hence are less appropriate to prime the flow paths because of the occurrence of gas bubbles in the tubing of the flow path. These pumps have check valves, usually balls located on both the intake and discharge sides; if these valves become clogged the pump will become inaccurate in its metering capability and will then lose suction / prime.

[0124] Peristaltic pumps are simple and easy to use and, they can handle thick or dirty liquids well, since there are no check valves to become clogged. Also, they operate more easily against higher back- pressures and thicker solutions and can be operated in a discontinuous manner. Peristaltic pumps also block the liquid when they are stopped. Finally, they prime easily and provide good suction capabilities for most liquids. Still, peristaltic pumps have drawbacks, mainly involving their hose. By incessant squeezing, the hose is steadily weakened until it deteriorates or also known as fatigue. Not only are the feed rate and pumping capability diminished over time, the pump motor itself is constantly under a load. Of course, this workload consumes more energy. Yet, the most serious downside of peristaltic pumps involves the possibility that the hose may rupture. Peristaltic pumps also generate more shear and allow lower flow rates than diaphragm pumps.

[0125] As such, both pump types have advantages and disadvantages and choice of the most optimal pump depends on the characteristics of the liquid and the volume to be added or removed.

[0126] In an embodiment, the unit comprises pump drives for disposable pump heads. In the latter case, the disposable pump heads may form part of a disposable flow path and can be received by the pump drives positioned on the unit.

[0127] The unit of the invention allows to choose a given flow path based on the specific requirements of the liquid addition / removal step. For instance, the unit of the invention allows the possibility to use small addition and / or large addition flow paths based on the process needs.

[0128] As such, in an embodiment, one or more of said flow paths can be designed for addition of small volumes, for instance volumes up to 10 L. In a further embodiment, said liquid flow in said flow path for small volume addition is controlled by a peristaltic pump.

[0129] In an embodiment, one or more of said flow paths can be designed for addition of larger volumes, for instance volumes starting from 10 L. In a further embodiment, said liquid flow in said large volume addition flow path is transported by a low shear pump (such as a diaphragm pump).

[0130] In a preferred embodiment, the unit of the current invention comprises at least one low shear pump such as a diaphragm pump and at least one peristaltic pump in order to efficiently add and remove multiple types of liquids to the bioreactor residing in said unit. In an embodiment, the unit of the current invention comprises one low shear pump. In an embodiment, the unit of the current invention comprises one low shear pump, for instance having an optimal pump flow rate range between 0.5-4 L / min (also referred to as "Large additions pump").

[0131] In an embodiment, the liquid flow in a given flow path is controlled by one pump. In another embodiment, the liquid flow in a given flow path is controlled by more than one pump, such as 2, 3, 4, 5 or 6 pumps.

[0132] In a preferred embodiment, the unit of the current invention comprises at least one low shear pump and at least 2 peristaltic pumps. In a preferred embodiment, the unit of the current invention comprises at least one low shear pump and at least 3, such as 4 peristaltic pumps.

[0133] In an embodiment, the unit of the current invention comprises at least 2 peristaltic pumps, one for pumping the cell medium in the bioreactor (also referred to as "BioIn pump") and one for pumping the cell medium out of the bioreactor (also referred to as "BioOut pump"). In a further embodiment, said 2 peristaltic pumps for pumping the cell medium in and out of the bioreactor have the same optimal pump flow rate range, for instance between 0.021 and 2.5 L / min.

[0134] In an embodiment, the unit of the current invention comprises one or more peristaltic pumps for pumping the cell medium in the bioreactor ( "BioIn pump"), one or more peristaltic pumps for pumping the cell medium out the bioreactor ( "BioOut pump") and one or more additional peristaltic pumps for adding liquid reagents to the bioreactor.

[0135] Said one or more additional peristaltic pumps can have the same or a different optimal pump flow rate range. In a further preferred embodiment, said one or more additional peristaltic pumps each have a different optimal pump flow rate range. In an embodiment, said unit comprises 2 additional peristaltic pumps, wherein each have a different optimal pump flow rate range. In an embodiment, said unit comprises 2 additional peristaltic pumps. In a further embodiment, said first additional peristaltic pump has an optimal pump flow rate range between 10 and 1000 mL / min, for instance 20-600 mL / min (also referred to as "small additions pump") and said second additional peristaltic pump has an optimal pump flow rate range between 0.1 and 100 mL / min, for instance 0.5 and 42 mL / min (also referred to as "Base pump").

[0136] In a preferred embodiment, the unit of the current invention comprises one low shear pump and 4 peristaltic pumps. In a further preferred embodiment, the unit of the current invention comprises one low shear pump and 4 peristaltic pumps, comprising one peristaltic pump for pumping the cell medium in the bioreactor ( "BioIn pump"), one peristaltic pump for pumping the cell medium out the bioreactor ("BioOut pump") and two additional peristaltic pumps for adding liquid reagents to the bioreactor.

[0137] The flow path can suitably comprise sanitary or aseptic connectors in order to allow for aseptic connection of the pre-sterilized flow path to pre-sterilized containers or one or more further flow paths. In an embodiment, a (pre-sterilized) flow path is connected to a (pre-sterilized) container by means of an aseptic sealing or by means of an aseptic tube welding.

[0138] Liquid reagents (for instance cell growth medium, an inoculum comprising cells, transfection reagent, lysis reagent, etc.) can be provided to the bioreactor by means of the plurality of flow paths, wherein said unit can comprise one or more aseptic connectors to connect one or more containers comprising a liquid reagent to a chosen flow path. Such a container can be any recipient known from the art and can include for instance bottles, (flexible) bags, tanks ... In a preferred embodiment, said container is a closed recipient. In an embodiment, said container comprises a singleuse (disposable) container, such as a single-use BioProcess container. In another embodiment, said container is reusable. In an embodiment, said container is flexible.

[0139] Examples of such containers are for instance transfer bags of 5 and 10 liters. Exemplary assemblies of such transfer bags are depicted in Figures 43 and 44.

[0140] In an embodiment, said unit further comprises one or more supports for installation of said one or more containers comprising a liquid reagent. Said supports can be any type of supports known from the art which allow installation of the containers, such as (adjustable) clamps or shelves. In a further embodiment, one or more of said supports are collapsible. In a further embodiment, one or more of said supports are removable. In a preferred embodiment, said supports are present on a front panel or on a lateral panel of said unit, not only increasing visibility of the containers, but also increasing accessibility, making the system more ergonomic for the operator when standing in front of the unit and facing the front panel.

[0141] In an embodiment, the invention provides a kit comprising a plurality of disposable (single-use) flow paths suited to be placed on the unit of the current invention. In an embodiment, each of said disposable flow paths is a closed flow path comprising a piping consisting of a single-piece tubing for making a fluid connection between a container comprising a liquid reagent and the bioreactor. In an embodiment, all flow path tubing can be replaced depending on a given process using dedicated automated phases.

[0142] In an embodiment, the kit further comprises other single-use components to be used with the unit of the current invention, such as one or more single-use containers for comprising a liquid reagent, one or more single-use probes or sensors, one or more single-use bioreactors, one or more single-use pumps, etc.

[0143] In an embodiment, the bioreactor is developed for single-use and comprises disposable pre-fitted manifolds, for instance pre-fitted manifolds for a top and bottom liquid bioreactor drain, a liquid sample line, a bubble or foam trap, a base addition line, etc.

[0144] In a further aspect, the invention relates to a method for executing a bioprocessing step in a bioprocessing unit, wherein said method can for instance comprising one or more of the following steps of: installing one or more flow paths comprising a plurality of disposable tubing on said unit by means of flow path guides; installing one or more containers of choice comprising a liquid reagent on one or more supports present on said unit; connecting each of said one or more containers with a flow path of choice; providing a controller with instructions for operating the liquid flow in said flow paths, wherein said controller is adapted to store a request sequence, and wherein said controller is configured to initiate said request sequence in an operating mode, thereby controlling the liquid flow in one or more flow paths by means of activation and / or deactivation of one or more pumps and / or valves connected to said one or more flow paths. Each of said one or more containers is connected with a flow path of choice. In an embodiment, the flow path is chosen depending on the request sequence such that when the request sequence is initiated, the liquid comprised in the container will follow the correct flow path, with the correct associated flow rate and timing of addition. In an embodiment, the request sequence is prepared and based on said request sequence, the one or more flow paths and / or one or more containers are installed.

[0145] In an embodiment, the unit comprises one or more sensors to indicate whether or not the correct components (for instance the correct containers) are installed on the correct position of the unit. In a further embodiment, the controller will only operate a request sequence if the right components are present.

[0146] In an embodiment, said bioprocessing step comprises the production of cells and / or the production of biomolecules such as a protein, a virus or viral particle, a cell or tissue therapy product or a gene therapy product.

[0147] In a further aspect, the invention relates to use of the unit as described above for executing a bioprocessing step, wherein said bioprocessing step comprises the production of cells and / or biomolecules such as a protein, a virus or viral particle, a cell or tissue therapy product or a gene therapy product.

[0148] In a further aspect, the invention relates to a method of installing a flow path comprising a plurality of disposable tubing on the unit as described above, said method comprising: choosing a suitable flow path and related pump, receiving said disposable tubing of the chosen flow path in guides positioned on the unit and connecting one end of said disposable tubing with a container of choice and the other end with the bioreactor. In an embodiment, said disposable tubing comprises multiple sections of tubing, wherein two consecutive sections of tubing are separated the one from the other by positioning of various components in between said sections of tubing (e.g. a container, a valve, an aseptic connector, a sensor, a split tubing assembly, etc.). In another embodiment, said disposable tubing consists of one uninterrupted single piece of tubing directly connecting a container of choice with the bioreactor.

[0149] In an embodiment, said unit further comprises one or more flow paths for liquid addition or removal without a pump, for instance by means of gravity. As such, the flow of liquid in said one or more flow paths is not controlled by means of a pump, but by means of gravity. As described above, said flow path can be connected to a container comprising the liquid reagent. In an embodiment, the liquid reagent can flow into the flow path by means of gravity, for instance by simply connecting the container to the flow path and opening a valve. In another embodiment, said liquid reagent is pumped from the container into the desired flow path.

[0150] In a preferred embodiment, the unit of the current invention comprises one low shear pump and 4 peristaltic pumps and one or more flow paths for liquid addition by means of gravity. In a further preferred embodiment, the unit of the current invention comprises one low shear pump and 4 peristaltic pumps, comprising one peristaltic pump for pumping the cell medium in the bioreactor ("BioIn pump"), one peristaltic pump for pumping the cell medium out the bioreactor ( and two additional peristaltic pumps for adding liquid reagents to the bioreactor and one flow path for liquid addition by means of gravity. In a further preferred embodiment, said first additional peristaltic pump has an optimal pump flow rate range between 20-600 mL / min ("small additions pump") and said second additional peristaltic pump has an optimal pump flow rate range between 0.5 and 42 mL / min ( "Base pump").

[0151] As described above, one or more request sequences can be chosen by an operator and instructed to the controller of the bioprocessing unit by means of a user interface.

[0152] In an embodiment, said user interface comprises an optical display comprising a graphical user interface, said optical display is able to present an outline of said flow paths to said operator. Said user interface allows the operator to provide the unit with instructions by selecting or imputing one or more request sequences. In an embodiment, the optical display is able to show the installation of the one or more flow paths and / or the one or more containers in real time. In an embodiment, the optical display is able to show the various components of the unit (and their status), such as the bioreactor, the pumps, the valves, the sensors, etc. In an embodiment, the guides and flow paths are visually reproduced in graphic format on the user interface.

[0153] In an embodiment, said optical display comprising said graphical user interface is positioned at a front panel of said unit. In an embodiment, said optical display comprising said graphical user interface is connected to a rotatable arm, allowing flexible positioning of said optical display. As such, this enables the operator to manipulate the interface while maintaining a view on the flow paths both located on the left (when front-facing) and front sides of the unit.

[0154] Such a request sequence can include a multitude a process steps, for instance the ones listed here below and visualized in figures 6-24. Any specific values regarding flow rate, the upper limit of fluid addition, temperature, pH, volume ranges of containers / bioreactors, etc. in these figures is merely exemplary and is not limitative for the unit and / or the process steps of the request sequence. Process values / steps as disclosed in these figures or their figure description are based on best effort estimation for common processes to be addressed by an embodiment of the unit / the method of the present disclosure. These request sequences listed below cannot be viewed as a list of exhaustive bioprocess possibilities of the unit / the method of the present disclosure.

[0155] Figure 5 shows a general overview of a flow diagram of the unit according to an embodiment of the present disclosure. The legend of the flow diagram is indicated in the right and is also applicable to figures 6-24. For instance, active ways in a certain request sequence and related flow diagram are indicated in grey.

[0156] In an embodiment (see figure 6), a request sequence comprises a step of filling of the bioreactor with (growth) medium by activating a peristaltic pump of the unit (option 1) (such as the "BioIn pump" described above, for instance allowing a flow rate of up to 2.5 L / min) connected to an influent flow path connected to a container comprising medium. In an alternative embodiment, filling of the bioreactor with (growth) medium can occur by instructing a low shear pump of the unit (option 2) (such as the "large additions pump" described above, allowing for instance a flow rate of up to 4.0 L / min or higher) connected to an influent flow path connected to a container comprising medium.

[0157] In an embodiment (see figure 7), a request sequence comprises a step of equilibrating the medium inside the bioreactor to obtain a stable pH, dissolved oxygen (DO) concentration and / or temperature. In a preferred further embodiment, said step occurs when the unit performs in batch mode (hence when no recirculation of perfusion of growth medium occurs). In an embodiment (see figure 7), a request sequence comprises a step of liquid sampling of the medium in the bioreactor after equilibration and before inoculation of the bioreactor with cells, for instance to perform offline measurements and / or to correct potential drifts of the sampling probes. Off-line measurements are performed by taking liquid samples from the bioreactor and analyzing these with other analyzer systems independent from the controller of the unit.

[0158] Examples of off-line measurements include:

[0159] - measurement of pH / DO to ensure proper calibrated status of probes set on the system (good practice);

[0160] - Measurement of media components such as glucose (key nutrients) and lactate (metabolic by product generated by growing cells) to ensure appropriate feeding of the culture and estimate cell numbers inside the bioreactor;

[0161] - Measurement of any other relevant components critical to ensuring optimal process conditions;

[0162] - Measurement of product titers, suspension turbidity, conductivity or other process related parameters to ensure proper process optimization

[0163] - sampling of the bed of the bioreactor, etc.

[0164] In an embodiment, the unit comprises an assembly enabling aseptic liquid sampling (see figure 28) for instance from the bioreactor or from the harvest tank. In an embodiment, the sampling assembly is single-use and hence reduces the risk of cross-contamination while delivering significant additional time and cost savings in the areas of assembly, cleaning, and cleaning validation. The sampling assembly can comprise any number of sampling tubes. In a preferred embodiment, the sampling assembly comprises between 2 and 12 sampling tubes, such as 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12 sampling tubes. In an embodiment, the sampling assembly further comprises a purge bottle, for instance at the end of the assembly. Said purge bottle can be of any size, for instance a 500mL purge bottle. Each individual sampling tube is connected to the assembly with tubing. In an embodiment, said connection further comprises a clamp. In an embodiment, each sampling tube comprises a container closure, a vent filter (for instance one of 0.2 pm), and / or a cap (for instance made from polyethylene). The sampling assembly is engineered to ensure an unobstructed fluid path, extremely low levels of extractables, and a secure elastomeric seal between closure and sampling tube. These assemblies are however relatively bulky and the number of tubes and filters composing them may impact customer experience when drawing liquid samples and aseptically disconnecting tubes.

[0165] As such, in an embodiment, the unit further comprises a holder designed to support the liquid sampling assembly and facilitate drawing of samples.

[0166] In an embodiment, said holder comprises one or more perforated plates supporting one or more sampling tubes and a top platform supporting and guiding the central tubing section of the sampling assembly (see Figure 32). Any number of sampling tubes and any size of sampling tubes can be envisioned. In an embodiment, the sample holder is adapted to receive 10 sampling tubes of either 15 or 50 mL. The sample holder can also be adapted to receive more or less than 10 sampling tubes and said tubes can be 15 or 50 mL or any other volume. By supporting and guiding all tubes and components of the assemblies, it greatly facilitates sampling as well as aseptic separation of tubes containing liquid samples.

[0167] In a preferred further embodiment, said step occurs when the unit performs in batch mode (hence when no external recirculation, but only recirculation inside the bioreactor occurs).

[0168] In an embodiment (see figure 8), a request sequence comprises a step of inoculation of cells in the bioreactor in batch mode, wherein the request sequence includes the removal of a volume equivalent to the inoculum volume by means of activation of a peristaltic pump (such as the "BioOut pump" described above) connected to an effluent flow path (a drain line) and subsequent filling of the bioreactor with an inoculum comprising the cells by means of a activation of a peristaltic pump (such as the "BioIn pump" or the "Small addition pump" described above) connected to an influent flow path or by means of a gravity controlled flow path. In a further embodiment, said gravity controlled flow path can be connected (using a pump) to a bottle or a (flexible) container. In an embodiment, the inoculum is comprised in a transfer bottle which can be connected to the unit. Use of a low shear pump (such as the "large addition pump" described above) is not recommended for the inoculation step as its use impacts the yield because of its larger residual volume, its lower accuracy, and its requirement to have a higher minimal volume to prime the pump. In an embodiment (see figure 9), a request sequence comprises a step of inoculation of the cells in recirculation mode, wherein a container comprising an inoculum is connected to a flow path (the recirculation loop) which is controlled by activation of a peristaltic pump directing liquid flow towards the bioreactor ("BioIn pump" described above) and activation of a peristaltic pump directing liquid flow away from the bioreactor ("BioOut pump" described above).

[0169] In an embodiment (see figure 10), a request sequence comprises a step of cell growth in batch mode, comprising a waiting time which allows cells to attach to the growth matrix (for instance a fixed bed) inside the bioreactor.

[0170] In an embodiment (see figure 10), a request sequence comprises a step of liquid sampling during batch mode to verify the attachment of the cells to the growth matrix and check the mortality rate of the cells which remained in suspension.

[0171] In an embodiment (see figure 11), a request sequence comprises a step of cell growth in recirculation mode during which media is recirculated, wherein a container (the recirculation tank) is connected to a flow path (the recirculation loop) which is controlled by activation of a peristaltic pump directing liquid flow from the recirculation tank towards the bioreactor ("BioIn pump" described above) and activation of a peristaltic pump directing liquid flow away from the bioreactor towards the recirculation tank ("BioOut pump" described above).

[0172] In an embodiment (see figure 11), a request sequence comprises a step of liquid sampling to estimate the cell growth (for instance through the glucose / lactate profiles) and / or to correct potential drifts of the probes during the cell growth in recirculation mode. In an embodiment, a request sequence comprises a step of liquid sampling to collect supernatant for cell culture media analysis testing, for cell growth estimation based on metabolites (for instance through the glucose / lactate profiles) and / or to correct potential drifts of the probes during the cell culture.

[0173] In an embodiment (see figure 12), a request sequence comprises a step of cell growth in perfusion mode during which fresh medium is continuously added and used medium is removed, wherein continuous addition of fresh media (for instance comprised in medium container) occurs by means of a flow path (the "perfusion in" line) which is controlled by activation of a peristaltic pump directing liquid flow towards the bioreactor ("BioIn pump" described above) and wherein used media is collected (for instance in a waste vessel) by means of a flow path (the "perfusion out" line) which is controlled by activation of a peristaltic pump directing liquid flow away from the bioreactor ("BioOut pump" described above).

[0174] In an embodiment (see figure 12), a request sequence comprises a step of liquid sampling to estimate the cell growth (for instance through the glucose / lactate profiles) and / or to correct potential drifts of the probes during the cell growth in perfusion mode.

[0175] In an embodiment (see figure 13), a request sequence comprises a step of cell growth in fed batch mode during which one or more steps of partial or complete emptying of the medium inside the bioreactor is followed by addition of an equal volume of fresh medium (media replacement step), wherein partial or complete emptying of the used medium inside the bioreactor occurs prior to addition of new medium by means of activation of a peristaltic pump (such as the "BioOut pump" described above) connected to an effluent flow path (a drain line) directing liquid away from the bioreactor towards a waste container and wherein cell growth is achieved with addition of new media (an equal volume as the one previously removed) to the bioreactor by activating a peristaltic pump of the unit (such as the "BioIn pump" described above, for instance allowing a flow rate of up to 2.5 L / min) connected to an influent flow path connected to a container comprising medium. In an alternative embodiment, filling of the bioreactor with (growth) medium can occur by instructing a low shear pump of the unit (such as the "large additions pump" described above, for instance allowing a flow rate of up to 4.0 L / min or higher) connected to an influent flow path connected to a container comprising medium. One or more of said media replacement steps can be performed depending on the bioprocessing step.

[0176] In an embodiment (see figure 13), a request sequence comprises a step of liquid sampling to collect supernatant for cell culture media analysis testing, for cell growth estimation based on metabolites (for instance through the glucose / lactate profiles) and / or to correct potential drifts of the probes during the cell culture.

[0177] In an embodiment (see figure 14), a request sequence comprises one or more washing cycles, wherein such a cycle comprises the following steps:

[0178] • draining of the medium inside the bioreactor by means of activation of a peristaltic pump (such as the "BioOut pump" described above) connected to an effluent flow path (a drain line) directing liquid away from the bioreactor towards a waste container, • subsequent filling of the bioreactor with medium by activating a peristaltic pump of the unit (such as the "BioIn pump" described above, for instance allowing a flow rate of up to 2.5 L / min) connected to an influent flow path connected to a container comprising medium or by activating a low shear pump of the unit (such as the "large additions pump" described above, for instance allowing a flow rate of up to 4.0 L / min or higher) connected to an influent flow path connected to a container comprising medium,

[0179] • subsequent agitation of the bioreactor for a time period,

[0180] • followed by a step of equilibrating the medium inside the bioreactor to obtain a stable pH, dissolved oxygen (DO) concentration and / or temperature in batch mode (hence when no recirculation of perfusion of growth medium occurs).

[0181] In an embodiment, a request sequence can comprise one or more washing cycles. In an embodiment, the number of cycles depends on the desired dead volume / void volume (for instance, the liquid retained within a fixed-bed matrix due to capillarity) inside the bioreactor. In an embodiment, the request sequence comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more than 10 washing cycles, preferably the request sequence comprises 1 or 2 washing cycles.

[0182] In an embodiment (see figure 14), a request sequence comprises a step of liquid sampling to verify the media composition and the efficiency of washing after said one or more washing cycles.

[0183] In an embodiment (see figure 15), a request sequence comprises a step of transfection or infection of the cells inside the bioreactor in batch mode, wherein the transfection step occurs quickly, preferably under 15 minutes. In a preferred embodiment, if the transfection / infection mix contains cells / virus sensitive to shear stress, the transfection / infection mix can be added in the bioreactor by means of a flow path controlled preferably by a low shear pump or by means of a flow path controlled by gravity and not by means of a flow path controlled by a peristaltic pump or the large volume pump. In an embodiment, a request sequence comprises a transfection cycle, wherein such a cycle comprises the following steps: a volume equivalent to the transfection / infection volume is optionally removed from the bioreactor through the drain line, by means of activation of a peristaltic pump (such as the "BioOut pump" described above) connected to an effluent flow path (a drain line) directing liquid away from the bioreactor towards a waste container, - the transfection / infection mix can be added in the bioreactor by activating a low shear pump of the unit (such as the "large additions pump" described above) connected to an influent flow path connected to a container comprising the transfection / infection mix or by means of a gravity controlled flow path connected to a container comprising the transfection / infection mix,

[0184] - agitation of the bioreactor at a lower agitation speed (for instance 0,5-1 cm / s),

[0185] - flushing of the flow path used for addition of the transfection / infection mix.

[0186] In some embodiments, agitation can occur in the bioreactor via an agitator. In some embodiments, an agitator can be a rotatable, non-contact magnetic impeller, a blade or screw agitation system, or an external circulation system. In some embodiments, the agitator can comprise a disk blade turbine, a curved blade turbine, an open blade fluid foil axial impeller, a turbine impeller with pitched blades, or a three-blade propeller. In a preferred embodiment, the bioreactor comprises a magnetic stirrer comprising magnets. In a further preferred embodiment, the magnetic stirrer controls a propeller located in the bioreactor. In a further embodiment, the propeller is single-use. In an embodiment, agitation can occur outside the bioreactor via an agitation device, which could for instance be positioned below the bioreactor. In some embodiments, the agitator or agitation device can provide agitation between 0 to 1000 rotations per minute (rpm), for instance between 0 and 450 rpm, such as 0, 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, 200, 210, 220, 230, 240, 250, 260, 270, 280, 290, 300, 310, 320, 330, 340, 350, 360, 370, 380, 390, 400, 410, 420, 430, 440, 450 rpm or any value in between. In a preferred embodiment, the rate of the agitator is controlled by means of the HMI.

[0187] In an embodiment, agitation can occur outside the bioreactor via a vibration device which could for instance be positioned below the bioreactor, for instance a vibration table. In an embodiment, the bioreactor is integrated into a docking station, which includes a vibration table.

[0188] In some embodiments, a bioreactor disclosed herein may comprise heating and / or cooling devices, designed to heat and / or cool the culture medium. In an embodiment, one or more or all containers in the biomolecule production system may comprise heating and / or cooling devices. In some embodiments, the heating device can be an electrical element, an electrical coil or any other heating means generally used in the field of cell culture, such as for example a thermostatically controlled double jacket. In an embodiment, the heating means are heating plates. In an embodiment, the unit disclosed herein comprises a heating support for the bioreactor. In a further embodiment, said heating support is comprised of heating plates. In an embodiment, the heating device comprises 7 elements. In a further embodiment, each element comprises a temperature sensor. In a further embodiment, each element comprises a temperature limiter. In a further preferred embodiment, the temperature limiter is set to 110°C. In some embodiments, cooling device may be any suitable cooling devices such as a Peltier element.

[0189] Transfection in recirculation is not desired but sometimes it might be necessary (given that during batch mode a too high concentration of transfection reagent in the bioreactor may have a cytotoxic effect). In recirculation mode, the volume is higher, which allows to have a lower concentration of transfection reagent in the bioreactor.

[0190] As such, in an embodiment (see figure 16), a request sequence comprises a step of transfection or infection of the cells inside the bioreactor in recirculation mode, wherein a container (comprising the transfection / infection mix) is connected to a flow path (the recirculation loop) which is controlled by activation of a peristaltic pump directing liquid flow from the container towards the bioreactor ("BioIn pump" described above) and activation of a peristaltic pump directing liquid flow away from the bioreactor towards the container ("BioOut pump" described above).

[0191] In an embodiment (see figure 17), a request sequence comprises a step of production of the desired biomolecule via recirculation (1) or via perfusion (2), batch and fed batch. If via recirculation, the recirculation tank can be used during the production and during harvest. In perfusion, this tank can be used to harvest the bioreactor and another one with fresh medium can be connected to the inlet of the bioreactor.

[0192] In an embodiment (see figure 17), a request sequence comprises a step of liquid sampling to estimate cells metabolites, virus content and correct potential drifts of the probes.

[0193] In an embodiment (see figure 18), a request sequence comprises a step of harvesting of the desired biomolecule if the latter is secreted by the cells. The final harvest can be performed with Bio Out pump to the Harvest tank. In an embodiment (see figure 19), a request sequence comprises a step of lysis of the cells if the desired biomolecule is not (fully) secreted or released by the cells. In an embodiment, the lysis buffer can be added in the bioreactor via the "large addition pump" or the "bioIN pump" (see above for details). In an embodiment, the bioreactor is drained prior to addition of the lysis buffer (see above for details regarding a draining step).

[0194] In an embodiment, the bioreactor is agitated after addition of the lysis buffer during a period. In an embodiment, the temperature is controlled during said agitation step. In an embodiment, draining of the bioreactor occurs after said agitation step. This can occur via the draining line and "BioOut pump" for harvest. In an embodiment, intermediate washing steps (see above) can be performed. In an embodiment, the lysis step is followed by a quenching (depending on the lysis buffer solution used), and one or more rinsing steps. In an embodiment, said lysis step is repeated, for instance once, twice or three, four, five, six, seven or more than seven times.

[0195] In an embodiment (see figure 20), a request sequence comprises one or more rinsing steps to recover void volume. Multiple rinsing cycles can be performed which can comprise the following steps:

[0196] 1. Filling the bioreactor with production medium and / or rinsing buffer via the "BioIn pump" (1) or "large addition pump" (2) (see above).

[0197] 2. Bioreactor agitation for a certain period.

[0198] 3. Draining the bioreactor to the harvest tank with "BioOut pump" (see above).

[0199] In an embodiment (see figure 21), a request sequence comprises a step of DO control. DO control is achieved in keeping total gas in overlay and stirring set within the defined range while varying O2 sparger flow rate.

[0200] In an embodiment (see figure 22), a request sequence comprises a step of temperature regulation of the bioreactor.

[0201] In an embodiment (see figure 23 and 24), a request sequence comprises a step of pH control of the bioreactor by means of CO2 and / or base addition regulation.

[0202] In an embodiment, the pH inside the bioreactor is adjusted by means of a base adjustment kit. In an embodiment, this base adjustment kit comprises two parts, a container (for instance a bag or bottle assembly, for instance depicted in Figure 42), containing the base (preferably NaOH), and a transfer assembly (for instance depicted in Figure 41) to connect to the bioreactor and optionally to fill the container. In an embodiment, the container comprises a bag and the bag assembly is hung on a hook of the unit. In a further embodiment, the bag assembly comprises a 5 L single-use bag. In an embodiment, the container comprises a bottle, for instance a 5 L bottle. In an embodiment, the unit comprises two such bottles, for instance one attached to each of the two side panels. In a preferred embodiment, the attachment of the container with the bioreactor comprises an aseptic connection. In an embodiment, the base is added to the bioreactor by means of a pump, such as peristaltic pump ("Base pump" described above).

[0203] In some embodiments, the unit can comprise one or more process controllers. In an embodiment, one or more process controllers are configured to control both the bioreactor and the unit. In some embodiments, the process controller is configured to control operations of a bioreactor and / or a unit for executing a bioprocessing step and can include a plurality of sensors, a local computer, a local server, a remote computer, a remote server, or a network. In some embodiments, the bioreactor and / or the unit can include one or more sensors, for example, a pH sensor, a dissolved oxygen sensor, temperature sensor (e.g., a thermocouple), flow rate sensor, gas sensor, level sensor, cell density sensor or any other sensor. In some embodiments, the process controller can be operational to control aspects of a product manufacturing process, and can be coupled to sensors disposed in the bioreactor and / or the unit, for example, to control the temperature, volume flow rate or gas flow rate into the bioreactor and / or the unit in real time. In an embodiment, the process controller is divided in two parts, namely a Programmable Logic Controller (PLC) and a Supervisory Control and Data Acquisition (SCADA). The PLC is the intelligence of the system and is connected to the sensors and the actuators. The PLC contains only data and no power. The SCADA is important for visualisation, data historian and audit trail. This SCADA system runs on a server that stores the data historians and supports the visualization. In an embodiment, information can also be visualized from a client tablet. In an embodiment, the client network can be connected directly to the server for remote access. In some embodiments, a process controller can include a Human-Machine Interface (HMI), such as a display, for example, a computer monitor, a smart phone app, a tablet app, or an analog display, that can be accessed by a user to determine the state of the system (based on the sensors comprised in the system) and to control the system by means of various actuators, such as pumps, valves, heaters and agitators. In some embodiments, the process controller can include an input, for example, a keyboard, a separated smart tablet, a key pad, a mouse, a touch pen or a touch screen, to allow a user to enter control parameters for controlling the operation of the bioreactor and the unit. In some embodiments, the process controller can control access to the bioreactor and / or the unit.

[0204] In some embodiments, the bioreactor disclosed herein can comprise and or contain sensors (or probes) for monitoring different parameters. In an embodiment, the sensors or probes can be electrically connected. In another embodiment, the sensors can be wireless. In another embodiment, the bioreactor comprises both electrically connected sensors and wireless sensors. In some embodiments, a sensor disclosed herein can be located in any compartment of a bioreactor disclosed herein. In some embodiments, sensors described herein can be a gas sensor (e.g. air, oxygen, nitrogen, or carbon dioxide), pH sensor, temperature sensor, cell density sensor, level sensor or dissolved oxygen (DO) sensor. In some embodiments, the sensors disclosed herein can measure amongst other things, biomass or cell density, the dissolved oxygen partial pressure, oxygen content, the pH value, the temperature, pressure, flow rate, level, certain concentrations of nutriments, such as lactate, glucose, glutamine, glutamate, ammonium or any metabolic product or product to be metabolized which could for example reflect the cell density. In some embodiment, cell density (biomass density) can be determined by electrical impedance analysis or electrical impedance spectroscopy using an arrangement of measuring electrodes or any other cell density measuring methods. During production of biomolecules, accurate determination of the volume, and hence the level of fluid, inside one or more of the vessels (such as the bioreactor) of the system is necessary. As such, in some embodiments, a bioreactor according to the disclosure can comprise one or more level sensors. In a preferred embodiment, the bioreactor comprises multiple level sensors positioned at different heights of the bioreactor. Such level sensors can aid in controlling the flow inside the bioreactor or can trigger an alarm in case of foam development inside the bioreactor. In an embodiment, one or more containers comprise one or more level sensors. Level sensors are known from the art and for instance use capacitive technology.

[0205] In an embodiment, a bioreactor according to the disclosure can comprise load cells. Load cells are force transducers configured to measure the weight of a vessel and its contents by detecting deformation of an internal strain gauge when subjected to mechanical load. By continuously monitoring the weight of the bioreactor, load cells enable indirect yet highly accurate measurement of the total fluid volume inside the vessel. This is particularly advantageous when non-invasive, real-time monitoring is desired, or in cases where conventional level sensors are limited due to vessel geometry, fluid properties (e.g., opacity or conductivity), or foaming behavior. Load cells may be integrated under the base of the vessel (e.g., as part of the support frame), and may be connected to a control unit that converts weight signals into volume readings based on known fluid density and vessel geometry. In some embodiments, the load cell data may also be used to regulate dosing, feeding, or harvesting operations, or to trigger alarms in the event of unexpected weight changes such as leaks, evaporation, or unplanned addition of fluid.

[0206] It will be clear to the person skilled in the art that these level sensors and / or load cells can also be used in relation to other vessels of the system.

[0207] In some embodiments, a bioreactor according to the disclosure can comprise sensors for measuring culture parameters. In some embodiments, a sensor disclosed herein can be in contact with culture medium in the bioreactor. In some embodiments, culture parameters can comprise amongst other things, the dissolved oxygen partial pressure, the pH, the temperature, the optical density, certain concentrations of nutriments, such as lactate, glucose, glutamine, glutamate, ammonium or any metabolic product or product to be metabolized which could for example reflect the cell density. In an embodiment, the part of the sensor which is placed into the bioreactor (for instance a pH probe) is single-use, whereas the part of the sensor not in contact with the bioreactor is multi-use (such as the transmitter of a pH sensor). In some embodiment, a bioreactor disclosed herein can use regulation loops according to the disclosed parameters. In some embodiments, a regulation loop can for example, modulate the quantity of oxygen to be injected according to the value of the dissolved oxygen partial pressure present or the quantity of dissolved oxygen consumed by the cells; speed of circulation of the culture medium; inject CO2 according to the pH value obtained by the sensors or any other type of regulation generally used in this type of culture. In some embodiments, cells can be exposed to dissolved oxygen concentrations of 50% of air saturation. In some embodiments, cells can be exposed to about 0%, 1%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 78%, 80%, 90%, or 100% nitrogen and / or about 0%, 1%, 5%, 10%, 21%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 110, 120, 130, 140 or 150% oxygen. In some embodiments, cells can be exposed to pure oxygen or an oxygen enriched atmosphere. In an embodiment, a sampling assembly can be connected to a bioreactor lid.

[0208] In an embodiment, information obtained from said sensors can be used to trigger an alarm when certain threshold values are surpassed. In an embodiment, one or more of said sensors can be deactivated during certain steps of a given request sequence, for instance: the level sensor can be deactivated during filling or emptying of the bioreactor.

[0209] Said unit is configured to comprise a bioreactor. In an embodiment, said unit does not comprise a bioreactor. In an embodiment, said unit comprises a bioreactor. In a further embodiment, said unit comprises a single-use bioreactor. In an embodiment, said single-use bioreactor comprises disposable pre-fitted manifolds for connecting with the flow path tubing of the unit (such as a for connecting with a drain line, a liquid sample line, a bubble or foam trap, etc.).

[0210] In an embodiment, the unit comprises pre-installed flow path tubing. In an embodiment, the unit does not comprise pre-installed flow path tubing. In an embodiment, the unit comprises pre-installed containers. In an embodiment, the unit does not comprise pre-installed containers.

[0211] In an embodiment, the disclosure provides a bioreactor cabinet configured to be incorporated in the unit of the current disclosure. In an embodiment, said bioreactor cabinet is preferably a wheeled (or otherwise mobile) bioreactor cabinet suited to receive a bioreactor. Such a bioreactor cabinet will allow to easily position and remove the bioreactor from the unit of the current disclosure, for instance when the cells inside the bioreactor would need to be harvested.

[0212] In a further embodiment, said bioreactor cabinet is provided thereto with a bioreactor docking station. In an embodiment, said bioreactor cabinet, more preferably a sidewall of said cabinet, is provided with a connector allowing the transmission of power, signals and / or data when paired with the unit of the current disclosure.

[0213] In an embodiment, said connector is able to introduce the coupling of the bioreactor cabinet to unit of the current disclosure by means of a connecting portion and receiving portion. Said connecting portion may be located at the bioreactor cabinet whereas the receiving portion is present in the unit or vice versa.

[0214] In an embodiment, the connector can be a modular connector system allowing combinations of power and signal contacts, Ethernet, optical fiber, coaxial contacts, hydraulic, pneumatic and thermocouplings in a compact frame or housing. This modular connector system can be configured according to the specific requirements of the connection. In a preferred embodiment the connectors are waterproof. In an embodiment, the male connector of the bioreactor cabinet is connected to the female connector of the production system. To ensure correct connection between the male and female connector, the female connector may contain centering pins. In another embodiment, the connector comprises an electronic eye to ensure correct connection. In another embodiment, the connector comprises magnetic elements to ensure correct connection.

[0215] In a further or in another embodiment, a connecting portion on the bioreactor cabinet and a receiving portion on the unit will allow docking of the bioreactor cabinet to the system to ensure that both entities are firmly connected to each other, prohibiting the release of the bioreactor cabinet from the system during the production of biomolecules. This connecting and receiving portion can be any known connecting system suitable in the art, such as of mechanical or magnetic system. A break-away function can be incorporated to be able to release the bioreactor cabinet from the system.

[0216] In some embodiments, culture medium can be circulated in the bioreactor via an agitator. In some embodiments, an agitator can be a rotatable, non-contact magnetic impeller, a blade or screw agitation system, or an external circulation system. In some embodiments, the agitator can comprise a disk blade turbine, a curved blade turbine, an open blade fluid foil axial impeller, a turbine impeller with pitched blades, or a three-blade propeller. In a preferred embodiment, the bioreactor comprises a magnetic stirrer comprising 5 magnets. In a further preferred embodiment, the magnetic stirrer controls a propeller located in the bioreactor. In a further embodiment, the propeller is single-use. In some embodiments, the agitator or agitation device can provide agitation between 0 to 1000 rotations per minute (rpm), for instance between 0 to 450 rpm, such as 0, 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, 200, 210, 220, 230, 240, 250, 260, 270, 280, 290, 300, 310, 320, 330, 340, 350, 360, 370, 380, 390, 400, 410, 420, 430, 440, 450 rpm or any value in between. In a preferred embodiment, the rate of the agitator is controlled by means of the HMI.

[0217] In an embodiment, said unit comprises a bioreactor docking station. In an embodiment, the docking station is removable or mobile. In a preferred embodiment, the bioreactor docking station resides (at least partially) inside the unit, preferably guarded from the outside environment by the walls of said unit. Consequently, the unit provides a protective shield to safeguard the bioreactor from harmful encounters. In an embodiment, the unit comprises a height adjuster to accommodate different bioreactor heights. In an embodiment, said height adjuster is positioned below the bioreactor.

[0218] In an embodiment, said bioreactor docking station comprises a height adjuster, for allowing positioning of said bioreactor. In an embodiment, said height adjuster is removable. As described above, in an embodiment, said bioreactor docking station and bioreactor are comprised in a bioreactor cabinet configured to be incorporated in the unit of the current disclosure.

[0219] By combining a chemical lysis step (using a lysis solution) with mechanical action target biomolecules are more efficiently recovered from the bioreactor. Such mechanical action or motion can be chosen from agitation / vibration, vortexing or moving a solution inside the bioreactor. In an embodiment, moving a solution comprises filling and / or draining the bioreactor with a solution. In an embodiment, said mechanical action or motion is selected from the group consisting of vibration / agitation, compaction, vortexing, rocking, applying ultrasound waves, filling and / or draining the bioreactor with a solution, expansion and combinations of the foregoing.

[0220] In an embodiment, the unit comprises an agitation / vibration device for agitating / vibrating the bioreactor. In a further embodiment, the unit comprises a fixation system for securing the bioreactor to the agitation / vibration device. In an embodiment, said agitation / vibration device comprises a deck adapted for receiving a bioreactor; and said fixation system comprising at least one fastener for securing the bioreactor to the deck. In an embodiment, the unit further comprises a bridge structure adapted for mounting to the bioreactor and receiving the at least one fastener.

[0221] In an embodiment, the bioreactor cabinet further comprises a vibration device / agitation device for vibrating / agitating the bioreactor. In a further embodiment, said vibration device / agitation device is positioned below the bioreactor, for instance between the height adjuster and the bioreactor.

[0222] Such height adjuster functions as a support to install the bioreactor. The height adjuster may be releasably attached to the unit and / or docking station and may vary in dimension, depending on the size and the dimension of the bioreactor. In an embodiment, the height adjuster will allow positioning of the bioreactor, regardless of its dimension, such that for example the top of the bioreactor is perfectly aligned with an opening provided in the unit for accessing the bioreactor or with connectors for connecting flow paths with the bioreactor. For example, the unit may be designed to fit a bioreactor of a specific size, such as a bioreactor with 600 m2internal growth surface. In case the same unit is to be used with a smaller bioreactor, e.g. a bioreactor of 200 m2internal growth surface, a height adjuster may be used to accommodate said smaller bioreactor. The latter allows ergonomic manipulation of said bioreactor and its outlets that are located on said surface. Said height adjuster may have any form suitable to be used, such as square, rectangular, round. In an embodiment, said height adjuster is a cylinder or disc-like element. The surface of the height adjuster can't allow deformation of the adjuster. The height adjuster may comprise any material suitable in the art such as plastic, aluminium, steel, metal alloy. Stainless steel has a high corrosion resistance and retains strength at high temperatures. In an embodiment, the height adjuster is adapted to change its position. The height adjuster might for instance be adapted to tilt, shake or rock or be lowered or elevated.

[0223] In some embodiments, the bioreactor can be a perfusion bioreactor, wave bioreactor, cylindrical bioreactor, bag bioreactor, moving bed bioreactor, packed bed bioreactor, fibrous bioreactor, membrane bioreactor, batch bioreactor, continuous bioreactor or combinations of the foregoing. In some embodiments, the bioreactors can be made from or comprise a suitable material, for example, stainless steel, glass, aluminum, or plastic or combinations thereof. In some embodiments, the bioreactor can allow for analysis of products.

[0224] In some embodiments, the bioreactor described herein comprises a fixed bed. In some embodiments, the fixed bed is a structured fixed bed (which means that it is formed of an easily replicated, generally homogeneous, substantially fixed structure, and thus is not randomly oriented or unstructured, and, as can be appreciated, could take a variety of sizes or shapes while meeting this qualification). In some embodiments, the structured fixed bed comprises a stack of substrate disks. The substrate layers of the disks are stacked with the first or second side of a substrate layer facing a first or second side of an adjacent substrate layer. In some embodiments, the structured fixed bed extends spirally around a tubular part. In some embodiments, the structured fixed bed described herein can provide for a large cell growth surface within a small volume while still allowing circulation of medium and cells. In some embodiments, the structured fixed bed can be a mesh or comprises a mesh structure. In some embodiments, mesh structure or mesh can be a structure comprising a network or web-like pattern of filament, wire or thread. In some embodiments, the network can define pores, openings or perforations formed of a three-dimensional weave. In some embodiments, the structured fixed bed described herein can comprise a tortuous path for cells and cell culture media. In some embodiments, the tortuous path or channel formed creates turbulence which facilitates cell and cell medium incursion into and / or through the structured fixed bed. In some embodiments the mesh structure is a cell immobilization structure. In some embodiments the mesh structure is or forms a spacer layer or section for flow of cells and medium. In some embodiments the mesh structure is both a cell immobilization and a spacer layer section. In an embodiment, said spacer sections may create equidistant spaces between layers for tissue, meat or seafood growth.

[0225] In some embodiments, a spacer layer facilitates the path for flow of cells and medium. In some embodiments, the structured fixed bed can comprise one or more cell immobilization layers having a surface which allows cells to adhere and grow upon and forming a cell immobilization section. In some embodiments, adjacent to the cell immobilization layers are one or more spacer layers. In some embodiments, the spacer layer can include a structure which forms a spacer section. In some embodiments, the spacer section allows passage of cells and medium through an open but tortuous path. In some embodiments, the structure or nature of the spacer layers can be chosen such that the spacer layers create a tortuous, open path for cells and culture media to travel in parallel to the surface of said spacer and cell immobilization layers. In some embodiments, the tortuous path or channel formed by the spacer section creates turbulence which facilitates cell and cell medium incursion into the immobilization layers.

[0226] In some embodiments, the spacer layer can be a mesh or comprises a mesh structure. In some embodiments, mesh structure or mesh can be a structure comprising a network or web-like pattern of filament, wire or thread. In some embodiments, the network can define pores, openings or perforations formed of a three-dimensional weave. In some embodiments, the spacer layers and / or the cell immobilization layers of a spacer section and a immobilization section can be made of a biocompatible material, preferably a biocompatible polymer, for example polyester, polyethylene, polypropylene, polyamide, plasma treated polyethylene, plasma treated polyester, plasma treated polypropylene or plasma treated polyamide. In some embodiments, the spacer layer or the cell immobilization layer can comprise silica, polystyrene, agarose, styrene divinylbenzene, polyacrylonitrile or latex. In some embodiments, the spacer layers and / or the cell immobilization layers of a spacer section and a immobilization section can be made of an edible material, such as chitin or cellulose. Cellulose has great potential as a scaffolding material for culturing of cells due to its versatility, biocompatibility. In an embodiment, cellulose and its derivatives are functionalized by mixing them with other materials to improve their chemical, physical or biological properties. Another equally popular substance that can be used as a scaffold is chitin, a polysaccharide found in the exoskeletons of arthropods and cell walls of fungi. It presents certain advantages as a scaffold material, as its fibrous structure provides mechanical support and mimics the texture and structure of muscle tissue. Chitin is also mostly biocompatible and can support the attachment, growth, and differentiation of cells for instance for meat tissue regeneration. In some embodiments, the layers can be hydrophilic or hydrophobic. In some embodiments, the cell immobilization layer can be hydrophilic. In some embodiments, a cell immobilization layer can be woven or nonwoven. In some embodiments, a cell immobilization section and a spacer section can be alternately positioned. In some embodiment, alternately positioned sections can alternate in a vertical position or in a horizontal position. In some embodiments, cell immobilization sections may be layered or alternately positioned in a vertical position or in a horizontal position. In some embodiments one or more layers may be connected. In some embodiments, one or more layers of cell immobilization layers can be superimposed on one or more spacer layers (or vice versa). In some embodiments, a structured bed disclosed herein can be tightly or loosely rolled to a structure such as a spiral structure, a monolith structure or varying shape or could be formed of layers one on top of one another with fluid flowing in parallel or perpendicular to the surfaces of the layers.

[0227] In some embodiments, the fixed bed growth surfaces may range from 0.1 to 2m2, 1 m2to 2 m2, 7-40 m2, 150-800 m2, and may vary among different sizes (height or diameter) of bioreactors. As noted, a plurality of fixed beds may be provided in a stacked configuration, such as one, two, three, four, or more fixed beds. In an embodiment, said fixed bed growth surface may for instance be 200 m2or 600 m2. Depending on the size of the fixed bed and / or the bioreactor, the unit of the current disclosure might be adapted. In an embodiment, the heating support or the agitation device will be adapted depending on the size of the bioreactor. In an embodiment, the docking station for the bioreactor and / or the bioreactor cabinet will be adapted depending on the size of the bioreactor. In an embodiment, said unit is a mobile system, preferably a wheeled system to allow transport. In some embodiments, the unit comprises tracks to allow transport. In an embodiment, the unit may be provided with structures or components that allow the mobility or transportation of said unit. Transportation means can include any means suitable in the art, both manually and / or electronically controlled, and include but are not limited to wheels, tracks, rolls.... Alternatively, or in addition to, in a further embodiment, said unit may be provided with suitable structures that allow conjunction with an elevator, conveyor or lifting device that may subsequently transport said unit. In a preferred embodiment, the unit comprises wheels.

[0228] The unit might be made of any material suitable in the art such as metal alloy, metal, or plastic. In one embodiment, the unit is made from a material comprising aluminum or stainless steel. In a specifically preferred embodiment, the unit is made of a material comprising stainless steel.

[0229] In an embodiment, the unit is designed to be installed close to a wall, for instance at a distance of less than 1 meter from a wall, such as at a distance of 20-40 cm from a wall. A certain distance from the wall is required for ventilation purposes and / or cleaning purposes. In an embodiment, the unit comprises stabilization means, such as feet, to allow proper stabilization during operation. In a further embodiment, the unit comprises 4 feet, one on each corner of the unit. In an embodiment, said one or more feet are adjustable in height, for instance to adjust the horizontally of the bioreactor. In an embodiment, the unit further comprises wheels, allowing the system to be moved. In a further embodiment, the unit comprises 4 wheels. In some embodiments, the wheels can be equipped with brakes and a directional lock. This will prevent unnecessary movement of the unit. In an embodiment, the one or more wheels are collapsible to avoid contact with the ground, for instance during operation of the unit. In an embodiment, when in operation, the unit is installed on feet, during which the wheels are collapsed and are not touching the ground.

[0230] In an embodiment, one or more components of the unit (the cabinet of the unit, the bioreactor cabinet, the bioreactor,..) are provided in submodules which can easily be dismantled and assembled back together. This allows to design a dismantlable system to help with shipping and installation.

[0231] In an embodiment, the unit according to the current disclosure can be combined with downstream processing equipment for in-line purification or clarification (for instance when the cells produce a secreted product). This downstream processing can occur in continuous, semi-continuous, or batch mode. In an embodiment, the unit according to the current disclosure can be combined with devices that allow further purification or filtration of the harvest of said bioreactor. The harvest can comprise of medium originating from the bioreactor or can be a lysate of the cells cultured in the bioreactor. Said purification means can be a combination of one or more of clarification, flocculation, precipitation of cell debris, lipids, host cell proteins, DNA, as well as ultrafiltration, tangential flow filtration aiming at concentrating the supernatant, or changing the chemical conditions (such as pH, conductivity, ionic strength). Said means can also be chromatographic means, in capture mode or in flow through mode; chromatography can be envisaged both in a packed mode, a monolith mode, a membrane-based mode or in a fluidized mode; should the chromatography be implemented in a fluidized mode, it can include the use of classical media separated by settling or centrifugation, or (para)magnetic media separated by an external magnetic field. It can be any combination of any of the means described previously. Such devices may include but are not limited to one or more chromatography column such as such as affinity chromatography, ionic exchange chromatography (e.g. anion or cation), hydrophobic interaction chromatography, size exclusion chromatography (SEC), immuno-affinity chromatography which is a column packed with an affinity resin, such as an anti-IgM resin, a Protein A, a Protein G, or an anti-IgG resin or any combination. The size of the column may vary based on the type of protein being purified and / or the volume of the solution from which said protein is to be purified.

[0232] In a further aspect, the invention relates to a method of operating a unit, said unit is configured to execute a bioprocessing step in a bioreactor residing in said unit, wherein said method comprises selecting one or more flow paths via a user interface of said unit according to a given request sequence, wherein said flow path can be an influent or an effluent flow path, directing a liquid towards or away from said bioreactor residing in said unit, wherein said user interface is connected to a controller, said controller stores said request sequence, which is initiated by said controller in the operating mode of said unit, thereby controlling the liquid flow in a given flow path by means of activation and / or deactivation of one or more pumps and / or valves of said unit.

[0233] As described above, in an embodiment, said request sequence can be preprogrammed and the operator can select such a preprogrammed request sequence via a user interface of the unit. In an embodiment, the operator can create his or her own request sequence by selecting one or more flow paths, for instance by means of the user interface.

[0234] The present invention will be now described in more details, referring to examples that are not limitative.

[0235] DESCRIPTION OF FIGURES

[0236] Figures 1-4 show a unit according to an embodiment of the current invention (a view of the left front side, the left side, the right front side and back of the unit in figures 1-4, respectively).

[0237] Figures 33-34 show a unit according to an embodiment of the current invention further displaying a container coupled to the flow path for small additions 72 and a container coupled to the flow path for addition by gravity 73.

[0238] The unit 1 comprises a plurality of flow paths realized by means of flow path tubing 2. Said flow paths can be influent or effluent flow paths, directing a liquid towards or away from a bioreactor 3. The unit comprises flow path guides 4 present on the front and side panels of the unit. Said flow path guides 4 are adapted to receive said flow path tubing 2. The guides allow to secure the flow path tubing 2 to the panels of the unit 1. As such, the guides 4 help restrain a disposable flow path tubing 2 with the desired slope. The disposable tubes 2 of the flow path can suitably be aligned with the guides 4, and the guides 4 can minimize any slack in flexible flow path tubes 2. During use, single use systems for treatment of bioprocess liquids require frequent installation and removal of the flow path tubing. This is greatly facilitated by the presence of guides 4. The flow paths are further visualized by means of laser engraving on the panels of the unit.

[0239] The unit further comprises a multitude of pumps, wherein each of said pumps is connected to a given flow path. The unit comprises one low shear pump 5 and 4 peristaltic pumps 6, 7, 8, 9 and a flow path for liquid addition by means of gravity. One peristaltic pump is used for pumping the cell medium in the bioreactor ("BioIn pump", 6), one peristaltic pump is used for pumping the cell medium out the bioreactor ("BioOut pump", 7). Said first additional peristaltic pump 8 has an optimal pump flow rate range between 20-600 mL / min ("small additions pump") and said second additional peristaltic pump 9 has an optimal pump flow rate range between 0.5 and 42 mL / min ("Base pump"). Both pump types have advantages and disadvantages and choice of the most optimal pump depends on the characteristics of the liquid and the volume to be added or removed. For instance, shear sensitive products (such as liquids comprising sensitive cells and lysates containing valuable proteins), should be pumped with a pump conferring a low shear 5, such as a diaphragm pump. Pinch valves 15 are equally provided to control the liquid flow in said flow paths.

[0240] Liquid reagents (for instance cell growth medium, an inoculum comprising cells, transfection reagent, lysis reagent, etc.) are provided to the bioreactor 3 by means of the plurality of flow paths. The unit 1 further comprises aseptic connectors to connect containers 11 comprising a liquid reagent to a chosen flow path.

[0241] The unit 1 further comprises supports for installation of said containers 11 comprising a liquid reagent. The supports are present on a front panel or on a lateral panel of said unit, not only increasing visibility of the containers, but also increasing accessibility, making the system more ergonomic for the operator when standing in front of the unit and facing the front panel. The unit further comprises a holder 16 designed to support the liquid sampling assembly and facilitate drawing of samples.

[0242] Request sequences can be chosen by an operator and instructed to the controller of the bioprocessing unit 1 by means of a user interface. The user interface comprises an optical display 12 comprising a graphical user interface, which is able to present an outline of the flow paths to the operator. The optical display 12 is positioned at a front panel of said unit 1 and connected to a rotatable arm 13, allowing flexible positioning of said optical display.

[0243] The unit further comprises a controller connected to the pumps and / or valves and is is adapted to store and initiate the request sequences as provided by the operator, thereby controlling the liquid flow in a given flow path by means of activation and / or deactivation of the pumps and / or valves of the unit.

[0244] The unit comprises wheels 14 which allow transportation of the unit and adjustable feet 15 for stabilization during operation.

[0245] The unit is a cabinet comprising side panels, front panels and back panels. The unit comprises an internal storage unit having multiple compartments which can be accessed by doors provided in the front (see figures 1 and 3) and back panels (see figure 4) of the unit. As depicted in Figure 2, the unit can further comprise a space for spare connectors 74, for instance on a side panel.

[0246] Such a space for spare connectors 76 can also be positioned on a front panel, see for instance Figure 34. Figure 34 further also depicts a foam trap 75 and a probe holder 79. An assembly for such a foam trap is depicted in Figure 38.

[0247] As depicted in Figures 30B and 34, signals for alarms and alerts can be generated by lights, such as LED lights 78. The unit can further be provided with lightning LEDs 77. These lightning LEDs ensure sufficient luminosity in the space above the bioreactor, as typically one must fulfill minimum lux requirements for proper operation. The LEDS intensity is tunable to adjust based on operating conditions.

[0248] Figures 5 shows a general overview of a flow diagram of the unit according to an embodiment of the present disclosure.

[0249] Figures 6-24 show flow diagrams of various request sequences according to an embodiment of the present disclosure. The various flow diagrams and request sequences are discussed above.

[0250] Figures 25-27 show tags assigned to one or more tubing supports to facilitate the installation of the tubing according to an embodiment of the present disclosure. The tags can be engraved on a panel of the unit (Figure 25), can be attached to the unit by means of an adhesive (Figure 26) or can comprises a LED system comprising LED lights (Figure 27) controlled by the controller of the unit according to an embodiment of the present disclosure.

[0251] Figure 28 depicts an assembly 17 enabling aseptic liquid sampling for instance from the bioreactor or from the harvest tank according to an embodiment of the present disclosure. The sampling assembly is single-use and hence reduces the risk of crosscontamination while delivering significant additional time and cost savings in the areas of assembly, cleaning, and cleaning validation. The sampling assembly comprises up to 10 sampling tubes 18. The sampling assembly 17 further comprises a 500 ml purge bottle 19 at the end of the assembly. Each individual sampling tube 18 is connected to the assembly with tubing and said connection further comprises a clamp 20 and an aseptic disconnection mechanisms 23. Each sampling tube comprises a container closure, a vent filter 21 and a cap 22. The sampling assembly is engineered to ensure an unobstructed fluid path, extremely low levels of extractables, and a secure elastomeric seal between closure and sampling tube.

[0252] Figure 29 depicts a holder 16 designed to support the liquid sampling assembly and facilitate drawing of samples according to an embodiment of the present disclosure. The holder comprises two perforated plates 23, 24 able to support 10 sampling tubes and a top platform 25 supporting and guiding the central tubing section of the sampling assembly. The sample holder is adapted to receive 10 sampling tubes of either 15 mL or 50mL. Adaptation to 50mL tubes can be done by replacing perforated plate 23 by an alternative design supporting 50mL tubes. By supporting and guiding all tubes and components of the assemblies, it greatly facilitates sampling as well as aseptic separation of tubes containing liquid samples.

[0253] Figure 30A and figure 31A show exemplary dimensions (expressed in mm) of a unit according to an embodiment of the present disclosure.

[0254] Figure 30B shows exemplary tags engraved on the front side of a unit according to an embodiment of the present disclosure. Box E for instance depicts engravings assisting in the positioning of containers used for base addition and addition of small volumes. Box F depicts an engraving assisting in the positioning of a drain line from the bioreactor. Signals for alarms and alerts can be generated by lights, such as LED lights 78. The unit can further be provided with lightning LEDs 77. Guides 87 allow correct placement of tubes in alignment with the flow path. Said guides 87 can be for instance black lines, colored lines or relief lines (for instance obtained by laser engraving and / or chemical etching) along which the disposable tubes of a flow path can be arranged. Said guides 87 can also be laser-engraved and / or chemically etched on one or more panels of said unit.

[0255] Figure 31B shows exemplary tags engraved on the left side of a unit according to an embodiment of the present disclosure. Box L for instance depicts engravings assisting in the positioning of containers used for collection of liquids drained from the bioreactor by means of the "BioOut pump", whereas box P depicts engravings assisting in the positioning of containers used for liquids which will be added to the bioreactor by means of the "BioIn pump". Box O depicts engravings assisting in the positioning of containers used for liquids which will be added to the bioreactor by means of the "large additions" pump. Figure 32 shows exemplary tags engraved on the right side of a unit according to an embodiment of the present disclosure. Box T for instance depicts engravings assisting in the positioning of holder for the sampling assembly. Box Q depicts engravings assisting in the positioning of a container used for addition of liquids to the bioreactor by means of gravity. Box S depicts engravings assisting in the connection of gas inlet tubings. Box R assists in the positioning of an venting gas line.

[0256] Figures 33-34 show schematic representations (left front side and right front side, respectively) of the unit according to an embodiment of the present disclosure also including containers connected to the flow path for small additions 72 and the flow path for gravity addition 73.

[0257] Figure 35 shows a side panel of the unit including manifolds according to an embodiment of the present disclosure. A container (for instance a flexible bag) can be provided on support 80. This container can for instance be coupled with the flow path for small additions. Details of an exemplary assembly for small additions are depicted in Figure 39. One peristaltic pump is used for pumping fluids towards the bioreactor ("BioIn pump", 6). This flow path further comprises tubing 84 and an assembly 82. Details of such an exemplary assembly are depicted in Figure 36. Another peristaltic pump (on a front panel, not shown) can be used for pumping fluids out the bioreactor (BioOut pump", 7). This flow path further comprises tubing 81 and an assembly 83. Details of such an exemplary assembly are depicted in Figure 37. Large additions can also be added to the bioreactor. This flow path further comprises tubing 86 and an assembly 85. Figure 40 shows such an exemplary assembly for large additions to the bioreactor. A pump for pumping large additions to the bioreactor can for instance be a diaphragm pump and can comprise a hardware component 88 installed on the unit and a single-use pump head that integrates in an assembly as shown in Figure 40 (component 54). Guides 87 allow correct placement of tubes in alignment with the flow path. Said guides 87 can be for instance black lines, colored lines or relief lines (for instance obtained by laser engraving, painting and / or chemical etching) along which the disposable tubes of a flow path can be arranged. Said guides 87 can also be laser-engraved and / or chemically etched on one or more panels of said unit.

[0258] Figure 36 shows an assembly for directing a liquid towards the bioreactor of the unit ("BIO IN") according to an embodiment of the invention. Numbering is as follows:

[0259] Figure 37 shows an assembly for directing a liquid away from the bioreactor of the unit ("BIO OUT") according to an embodiment of the invention. Numbering is as follows:

[0260] Figure 38 shows an assembly for a foam trap of the unit according to an embodiment of the invention. Numbering is as follows:

[0261]

[0262] Figure 39 shows an assembly for small additions to the bioreactor according to an embodiment of the invention. Numbering is as follows:

[0263] Figure 40 shows an assembly for large additions to the bioreactor according to an embodiment of the invention. Numbering is as follows:

[0264] Figure 41 shows a base assembly of the unit according to an embodiment of the invention. Numbering is as follows:

[0265] Figure 42 shows a base bottle assembly of the unit according to an embodiment of the invention. Numbering is as follows: Figure 43 shows a transfer bag assembly (5 liters) of the unit according to an embodiment of the invention. Numbering is as follows:

[0266] Figure 44 shows a transfer bag assembly (10 liters) of the unit according to an embodiment of the invention. Numbering is as follows:

[0267] The present invention is in no way limited to the embodiments described in the examples and / or shown in the figures. On the contrary, methods according to the present invention may be realized in many different ways without departing from the scope of the invention.

Claims

1. CLAIMS1. A unit for executing a bioprocessing step, wherein said unit comprises: at least one flow path, wherein the at least one flow path can be an influent or an effluent flow path, directing a liquid towards or away from a bioreactor residing in said unit;- at least one pump and / or at least one valve, wherein the at least one pump and / or valve is connected to the at least one flow path;- a controller, connected to said at least one pump and / or valve; a user interface, connected to the controller for allowing an operator to select the at least one flow path according to a given request sequence; wherein the controller is adapted to store said request sequence and wherein said controller is configured to initiate said request sequence, thereby controlling the liquid flow in the at least one flow path by means of activation and / or deactivation of said at least one pump and / or valve.

2. Unit according to claim 1, wherein said unit comprises one or more aseptic connectors to connect one or more containers comprising a liquid reagent to a chosen flow path.

3. Unit according to any of the previous claims, wherein said at least one pump is chosen from a low shear pump and a peristaltic pump.

4. Unit according to any of the previous claims, wherein said pumps are chosen from low flow rate pumps and high flow rate pumps.

5. Unit according to any of the previous claims, further comprising one or more flow paths for liquid addition by means of gravity.

6. Unit according to any of the previous claims, wherein said request sequence is preprogrammed and / or obtained via the user interface.

7. Unit according to any of the previous claims, wherein the unit is a cabinet, said cabinet comprising one or more side panels, one or more front panels and one or more back panels.

8. Unit according to any of the previous claims, wherein said user interface comprises an optical display comprising a graphical user interface, said optical display is able to present an outline of said flow paths to said operator.

9. Unit according to claim 8, wherein said optical display comprising said graphical user interface is positioned at a front panel of said unit.

10. Unit according to claim 9, wherein said optical display comprising said graphical user interface is connected to a rotatable and / or retractable arm, allowing flexible positioning of said optical display.

11. Unit according to any of the previous claims, wherein at least part of said influent flow paths are positioned such that they provide a slope of between 1.0 and 3.0 degrees from a horizontal plane.

12. Unit according to any of the previous claims, wherein at least part of said influent flow paths are uninterrupted paths.

13. Unit according to any of the previous claims, wherein one or more of said flow paths can be designed for addition of volumes up to 10 L, said liquid flow in said flow path being controlled by a peristaltic pump.

14. Unit according to any of the previous claims, wherein one or more of said flow paths can be designed for addition of volumes starting from 10 L, said liquid flow in said flow path being controlled by a diaphragm pump.

15. Unit according to any of the previous claims, wherein at least part of said flow paths comprise one or more sensors and / or indicators for measuring and indicating parameters such as flow rate, pressure and / or temperature.

16. Unit according to any of the previous claims, wherein flow path guides are present on one or more panels of said unit.

17. Unit according to claim 16, wherein said guides comprise visually and / or tactilely distinguishable lines for the visualization of said flow path.

18. Unit according to claim 17, wherein said guides are visualized on one or more panels of said unit.

19. Unit according to claim 17, wherein said guides are adapted to receive said flow path.

20. Unit according to claim 19, wherein said guides adapted to receive said flow paths are positioned on one or more of said panels in order to prevent crossing of one or more of said flow paths, increase visibility of the liquid flow in one or more flow paths and / or increase efficiency during operation and / or maintenance of the unit.

21. Unit according to any of the previous claims 2-20, wherein said unit further comprises one or more supports for installation of said one or more containers comprising a liquid reagent and wherein said supports are present on a front panel or on a lateral panel of said unit.

22. Unit according to any of the previous claims, wherein said bioreactor is a single-use bioreactor.

23. Unit according to any of the previous claims, wherein said unit comprises a bioreactor docking station.

24. Unit according to claim 23, wherein said bioreactor docking station comprises a height adjuster, for allowing positioning of said bioreactor.

25. Unit according to any of the previous claims, wherein said bioreactor comprises a fixed bed for culturing cells.

26. Unit according to claim 25, wherein said fixed bed is a structured fixed bed, optionally comprising a spiral bed.

27. Unit according to any of the previous claims 25-26, wherein said fixed bed has a surface of between 10 to 800m2, more preferably 200m2to 600m2.

28. Unit according to any of the previous claims, wherein said unit is a mobile system, preferably a wheeled system to allow transport.

29. Unit according to any of the previous claims, comprising an internal storage unit having one or more compartments.

30. Unit according to claim 29, wherein the one or more compartments can be accessed by doors provided in one or more front panels, side panels and / or back panels of the unit.

31. Unit according to any of the previous claims, wherein the unit is adapted to be juxtaposed against a second unit or wall.

32. Unit according to claim 31, wherein the back panel of the unit is adapted to be positioned against a wall.

33. Unit according to any of the previous claims, further comprising one or more probes for sampling the bioreactor.

34. Unit according to any of the previous claims, wherein one or more components of the unit, such as the cabinet or the bioreactor, are modular.

35. A method for executing a bioprocessing step in a bioprocessing unit, said method comprising the steps of: installing one or more flow paths comprising a plurality of disposable tubing on said unit by means of flow path guides; installing one or more containers of choice comprising a liquid reagent on one or more supports present on said unit; connecting each of said one or more containers with a flow path of choice; providing a controller with instructions for operating the liquid flow in said flow paths, wherein said controller is adapted to store a request sequence, and wherein said controller is configured to initiate said request sequence in an operating mode, thereby controlling the liquid flow in one or more flow paths by means of activation and / or deactivation of one or more pumps and / or valves connected to said one or more flow paths.

36. Method according to claim 34, wherein said bioprocessing step comprises the production of cells and / or the production of biomolecules such as a protein,a virus or viral particle, a cell or tissue therapy product or a gene therapy product.

37. Use of a unit according to the previous claims 1-33 for executing a bioprocessing step, wherein said bioprocessing step comprises the production of cells and / or the production of biomolecules such as a protein, a virus or viral particle, a cell or tissue therapy product or a gene therapy product.

38. A method of installing a flow path comprising a plurality of disposable tubing on the unit of any of the previous claims 1-33, comprising: choosing a suitable flow path and related pump, receiving said disposable tubing of the chosen flow path in guides positioned on the unit and connecting one end of said disposable tubing with a container of choice and the other end with the bioreactor.

39. A method of operating a unit, said unit is configured to execute a bioprocessing step in a bioreactor residing in said unit, wherein said method comprises selecting one or more flow paths via a user interface of said unit according to a given request sequence, wherein said flow path can be an influent or an effluent flow path, directing a liquid towards or away from said bioreactor residing in said unit, wherein said user interface is connected to a controller, said controller stores said request sequence, which is initiated by said controller in the operating mode of said unit, thereby controlling the liquid flow in a given flow path by means of activation and / or deactivation of one or more pumps and / or valves of said unit.

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