Cell culture medium specification attribute assessment

Abstract

The invention particularly discloses a method. The method comprises the following steps: obtaining the impurity level of at least one impurity of each raw material in a plurality of raw materials; obtaining a formula of the cell culture medium, wherein the formula shows the proportion of various raw materials in the cell culture medium; and determining a total impurity level of the at least one impurity in the cell culture medium based at least in part on the formulation and each impurity level.

Description

TECHNICAL FIELD

[0001] According to the present disclosure, various exemplary embodiments relate to the field of cell culture, in particular to the preparation of biological growth media (often referred to as cell culture media) for cell culture. More specifically, according to the present disclosure, various exemplary embodiments relate to a computer-implemented method for determining an impurity level of at least one impurity in a cell culture medium. BACKGROUND

[0002] In vitro cultivation of cells and tissues is one of the foundations of modern biotechnology. Providing reliable and efficient cell culture conditions helps to improve the production of cell culture products, such as biologies, e.g. therapeutic or diagnostic proteins, as well as reliable diagnostic tests that rely on cell culture. Achieving the best quality of cell culture depends on the synergy between the cells to be cultivated and the cell culture medium in which these cells are suspended. Therefore, it becomes increasingly important that cell culture media can be customized according to the specific cell line or even the product to be produced. Accordingly, the need for precise monitoring and control of the composition of cell culture media is also growing.

[0003] Cell processes often lack reproducibility, partly due to variability of cell culture media. One approach to address this issue is to replace any undefined compound, such as an animal, yeast or plant-derived compound, with a well-defined compound, such as a recombinantly produced protein. A typical feature of these chemically defined media is a significant reduction in process variability in cell culture processes performed with them. However, chemically defined media still suffer from variability, as individual compounds introduce impurities into the cell culture medium, which cumulatively affect the cell culture process. Furthermore, the impurity profile of each of these compounds varies from supplier to supplier and batch to batch, so there is a risk of reproducibility for each new source of compound. While some approaches in the art aim to reduce the impurities introduced into the cell culture medium along with the compounds, the purification and production processes of these purer compounds make these alternative approaches more cumbersome and therefore more costly. SUMMARY

[0004] According to the present disclosure, a computer-implemented method is disclosed, wherein the method comprises: - obtaining an impurity level of at least one impurity for each of a plurality of raw materials; - obtaining a recipe for a cell culture medium, wherein the recipe shows proportions of the plurality of raw materials in the cell culture medium; and - determining a total impurity level of the at least one impurity in the cell culture medium based at least in part on the recipe and the impurity levels.

[0005] Further, according to the present disclosure, a computer program is disclosed, wherein the computer program comprises instructions, which, when executed by a computer, cause the computer to perform the method according to the present disclosure. For example, the computer program comprises program instructions, which, when the computer program is run on a processor, cause the processor to perform and / or control the method according to the present disclosure. For example, the processor is to be understood to include, but is not limited to, a control unit, a microprocessor, a microcontroller unit (such as a microcontroller), a digital signal processor, an application-specific integrated circuit, or a field programmable gate array. In this case, all steps of the method can be controlled or all steps of the method can be performed or one or more steps can be controlled and one or more steps can be performed. The computer program can be distributed, for example, over a network, such as the Internet, a telephone or mobile radio network, and / or a local area network. The computer program can be at least partially a software and / or firmware of the processor. It can likewise be implemented at least partially as hardware. The computer program can be stored, for example, on a computer-readable storage medium, such as a magnetic, electric, optical, and / or other type of storage medium. The storage medium can be, for example, part of the processor, such as a (non-volatile or volatile) program memory of the processor or a part thereof. The storage medium can be, for example, a tangible or physical storage medium.

[0006] Further, according to the present disclosure, an apparatus is disclosed, wherein the apparatus is configured to perform and / or control the method according to the present disclosure. Alternatively, the apparatus can comprise devices (such as computer devices) for performing and / or controlling the individual steps of the method according to the present disclosure. In this case, all steps of the method can be controlled or all steps of the method can be performed or one or more steps can be controlled and one or more steps can be performed. One or more steps can also be performed and / or controlled by the same unit. For example, one or more steps can be performed by one or more processors.

[0007] Further, according to the present disclosure, an apparatus is disclosed, wherein the apparatus comprises at least one processor and at least one memory including program code, wherein the at least one memory and the program code are configured to, with the at least one processor, cause the apparatus at least to perform and / or control the method according to the present disclosure. In this case, all steps of the method can be controlled or all steps of the method can be performed or one or more steps can be controlled and one or more steps can be performed.

[0008] For example, a cell culture medium is understood to be a solid or liquid composition comprising or consisting of a plurality of starting materials. In a further example, a cell culture medium can contain a solvent, such as water, in addition to the plurality of starting materials. For example, a starting material is understood to be a component of a cell culture medium, which can be, for example, a compound, in particular an organic or inorganic compound, an inorganic salt, an organic salt, a single amino acid or a mixture of a plurality of different amino acids, a peptide, a protein, a sugar or a mixture of a plurality of sugars, a sugar alcohol, an oligosaccharide or a polysaccharide or a mixture of a plurality of oligosaccharides and / or polysaccharides, a nucleic acid or a mixture of a plurality of nucleic acids, an amino acid derivative, a bioamine, a vitamin, a fatty acid, a buffer, a dye or a biological substance (such as serum, for example fetal bovine serum (FBS)). For example, a starting material can be present in solid (such as a powder) or liquid form. A starting material can be, for example, fluorescently or radioactively labeled. As further described below, a starting material can contain one or more impurities.

[0009] For example, an impurity is understood to be a substance present in a starting material and can not be the specified compound of the starting material. Exemplary types or classes of impurities in each starting material of a cell culture medium include elemental impurities, inorganic compounds, organic compounds, endotoxins or biological load. Elemental impurities can refer, inter alia, to chemical elements. In a further example, impurities, inter alia but not limited to elemental impurities, can be toxic impurities and / or functional impurities. An impurity can be, for example, a solvent or a compound derived from a solvent. An impurity can also be referred to as a trait, a contaminant, a defect or similar expressions, for example.

[0010] For example, in the context of impurities, the terms “chemical element” and “element” are understood to be a chemical substance defined by its atomic number. In the present disclosure, the element symbols in the periodic table of elements can be used to refer to any charged, uncharged or complexed form of an element. For example, an elemental impurity can exist in the form of uncharged elemental particles, salts, ions, complexes, coordination compounds, etc.

[0011] For example, a toxic impurity is understood to be an elemental impurity which, when present in a cell culture medium in a certain amount or respective concentration (e.g. in an amount or concentration which exceeds a certain limit value), can have an adverse effect, such as a reduced yield of a cell culture product, a slowed cell growth, cell death, contamination of a cell culture product, a change in a quality attribute of a product, etc. For example, it can be preferred that the amount of toxic impurities in a cell culture medium is below an upper limit. Without being limited thereto, a toxic impurity can be, for example, Pb, Hg, As, Cd, Ag, Cr, Li, Sb or Ba, or a combination thereof. The term “a” (as in “toxic impurity”) as used herein is understood to mean “one or more”, unless the context explicitly dictates otherwise.

[0012] For example, a functional impurity is to be understood as an elemental impurity which, when present in a cell culture medium in a specific amount or respective concentration (e.g. in an amount or concentration above a lower limit and / or below an upper limit), can be beneficial for the cell culture and / or can even be at least partially essential for a normally functioning cell culture. In a further example, a functional impurity can cause adverse effects as described above for a toxic impurity when present, for example, in a specific amount or respective concentration above an upper limit or below a lower limit. Thus, it can be preferred, for example, that the functional impurity is present in the cell culture medium in an amount or concentration above a lower limit and below an upper limit. Without being limited thereto, the functional impurity can be, for example, V, Se, Ca, K, Mg, P, S, Sn, Na, Co, Fe, Cu, Mn, Mo, Ni or Zn, or a combination thereof.

[0013] For example, in the context of an impurity, an organic compound is to be understood as an organic chemical compound which, when present in a cell culture medium in a specific amount or respective concentration (e.g. in an amount or concentration exceeding a specific limit), can cause adverse effects such as a decrease in cell culture product yield, a decrease in cell growth, cell death, contamination of the cell culture product, a change in product quality attributes, etc. For example, it can be preferred that the amount of organic compound in the cell culture medium is below an upper limit. The organic compound can be, for example, dimethyl sulfoxide (DMSO), acetonitrile, methanol, ethanol, formaldehyde, glycerol, polyethylene glycol, etc.

[0014] For example, in the context of an impurity, an inorganic compound is to be understood as an inorganic chemical compound which, when present in a cell culture medium in a specific amount or respective concentration (e.g. in an amount or concentration exceeding a specific limit), can cause adverse effects such as a decrease in cell culture product yield, a decrease in cell growth, cell death, contamination of the cell culture product, a change in product quality attributes, etc. For example, it can be preferred that the amount of inorganic compound in the cell culture medium is below an upper limit. The inorganic compound can be, for example, an inorganic salt such as copper sulfate, zinc chloride or silver nitrate, arsenic chloride, antimony chloride, or an inorganic acid or base such as hydrochloric acid, sulfuric acid, sodium hydroxide, potassium hydroxide, etc.

[0015] For example, an endotoxin is to be understood as a chemical or biological by-product in a raw material production process (e.g. a production process performed in a biological organism). Since endotoxins are generally not beneficial or even harmful for a cell culture, it can be preferred that the amount or concentration of endotoxins in the cell culture medium does not exceed an upper limit. Without being limited thereto, the endotoxin can be, for example, at least one lipopolysaccharide, or at least one bacterial toxin.

[0016] For example, a biological load is to be understood as any archaea, prokaryote, eukaryote, or viral biological unit present in the raw materials or in the cell culture medium. Since a biological load can be unbeneficial or even harmful to the cell culture, it can be preferred, for example, that the amount of biological load in the cell culture medium does not exceed an upper limit. Without being limited thereto, the biological load can be, for example, a mycoplasma, a coccus, a yeast, a fungus, or a bacillus.

[0017] For example, an impurity level is to be understood as an amount or concentration of a respective impurity in the raw materials and / or in the cell culture medium. For example, an impurity level of a certain impurity in the cell culture medium can be determined at least partially based on an impurity level of said impurity of one or more raw materials comprised by the cell culture medium.

[0018] Obtaining the respective impurity levels or the recipe of the cell culture medium can be understood, for example, as meaning that information, parameters, and / or data indicative of the respective impurity levels or the recipe can be received (e.g., in a suitable electronic format such as a text format, a spreadsheet format, or a database format) at the device performing the method (e.g., via a communication interface of the device, e.g., from another device providing a database of raw materials or a database of recipes). For example, the respective impurity levels or the recipe of the cell culture medium can be requested to trigger or instruct such receiving. In another example, obtaining the respective impurity levels can comprise requesting another device to perform a determination and subsequently transmitting the specific impurity levels to trigger the determination of at least one impurity level.

[0019] Determining the total impurity level of the at least one impurity in the cell culture medium based at least partially on the recipe and the respective impurity levels can be understood, for example, as meaning that the total impurity level can be calculated or computed, wherein such calculation or computation can at least partially depend on the recipe and the respective impurity levels.

[0020] For example, the plurality of raw materials can be understood as the raw materials used for preparing the cell culture medium and comprised in the cell culture medium after the preparation of the cell culture medium has been completed. For example, the cell culture medium can comprise at least 20 to 30 or at most 150 raw materials, but the method of the present disclosure can be applicable to cell culture media containing any number of raw materials (e.g., at least 2, at least 3, at least 4, at least 5, at least 10, at least 20, or at least 30 raw materials). The proportions of the plurality of raw materials comprised in the cell culture medium can be indicated by the recipe of the cell culture medium. For example, the plurality of raw materials can correspond to all raw materials or a subset of all raw materials comprised in the cell culture medium.

[0021] For example, a formulation of a cell culture medium (such as a recipe of a cell culture medium) can be understood to be determined based on a specific weight ratio, molar ratio, or concentration of the mass or molar amount / volume of the plurality of starting materials contained in the cell culture medium. In some examples, the formulation can indicate the proportions of the plurality of starting materials in a relative manner (e.g., showing that a starting material A is contained in the cell culture medium at 20% and a starting material B is contained in the cell culture medium at 80%) or in an absolute manner (e.g., showing absolute amounts of the starting material A and the starting material B contained in the cell culture medium). Herein, for example, it can be preferred that the percentages correspond to weight percentages, e.g., that a content of a starting material in the cell culture medium at 20% corresponds to 20% by weight, wherein 100% or 100% by weight can correspond to the dry weight of the cell culture medium or the weight of the cell culture medium before hydration. For example, a formulation of a cell culture medium can refer to a specific amount of the cell culture medium. The formulation can be obtained, for example, by obtaining (e.g., receiving or determining) information, parameters, and / or data indicative of the formulation (e.g., in a suitable electronic format such as in a text format, a spreadsheet format, or a database format).

[0022] A level of an impurity in a starting material can be understood, for example, as the amount of that specific impurity in the starting material (e.g., in a specific amount of the starting material). For example, a starting material can comprise a plurality of impurities, the respective amounts of which are given by respective levels of impurities. The level of an impurity can be obtained, for example, by obtaining (e.g., receiving or determining) information, parameters, and / or data indicative of the level of the impurity (e.g., in a suitable electronic format such as in a text format, a spreadsheet format, or a database format).

[0023] A level of an impurity can be expressed in various ways. In one example, a level of an impurity can be expressed as parts per million (ppm), which indicates the number of parts of the impurity per million parts of the starting material. In another example, a level of an impurity can be expressed as a weight percentage, which indicates the weight of the impurity as a percentage of the total weight of the starting material. In another example, a level of an impurity can be expressed as a mole fraction, which indicates the ratio of the number of moles of the impurity to the number of moles of the starting material.

[0024] A total level of an impurity in a cell culture medium can be understood, for example, as the amount of that specific impurity in the cell culture medium (e.g., in a specific amount of the cell culture medium). For example, a cell culture medium can comprise a plurality of impurities, the respective amounts of which are given by respective total levels of impurities. Upon determination of a total level of an impurity, the total level of the impurity can be indicated, for example, by corresponding information, parameters, and / or data (e.g., in a suitable electronic format such as in a text format, a spreadsheet format, or a database format).

[0025] The total impurity level of the at least one impurity in the cell culture medium can be influenced by the impurity level of each of the at least one impurity in each of the plurality of raw materials of the cell culture medium. In other words, the total impurity level can be determined from the respective impurity levels of the at least one impurity of each of the raw materials and the proportions of the plurality of raw materials in the cell culture medium as indicated by the recipe. The total impurity level can then be understood as the total amount of the impurity in the cell culture medium, given as the sum of the respective amounts of the impurity of each of the raw materials.

[0026] Advantageously, by determining (e.g., calculating or computing) the total impurity level of the at least one impurity in the cell culture medium, the selection of the one or more raw materials contained in the cell culture medium can be optimized. For example, depending on whether the determined total impurity level meets a predefined criterion, it can be determined whether the selection of the raw materials needs to be optimized (e.g., by replacing one or more of the raw materials of a particular batch with another batch of the respective raw material having different respective impurity levels of the at least one impurity; or, for example, by replacing one or more of the raw materials with respective equivalent raw materials having different counter-ions and / or different hydration levels) to change the total impurity level in the cell culture medium. Since the determination of the total impurity level depends on the recipe and the respective impurity levels of each of the plurality of raw materials, in assessing the selection of the raw materials, the contribution made by each of the raw materials can be considered in terms of its content in the cell culture medium. For example, in embodiments comprising replacing one or more of the raw materials with respective equivalent raw materials having different counter-ions, it can be preferred that the counter-ions do not significantly change the respective elemental content in the composition, or, preferably, are balanced by a corresponding change in another raw material, so that the ionic content in the cell culture medium remains unchanged.

[0027] Next, further exemplary features and exemplary embodiments according to the present disclosure will be described in more detail.

[0028] According to exemplary embodiments, the method of the present disclosure further comprises: - determining at least one other total impurity level of at least one other impurity in the cell culture medium based at least in part on the recipe and the respective impurity levels of the at least one other impurity of each of the plurality of raw materials.

[0029] For example, the at least one other impurity can be selected from the group consisting of elemental impurities, inorganic compounds, organic compounds, endotoxins, and biological load, independently of the at least one impurity.

[0030] For example, at least one further impurity is to be understood as comprising any number of further impurities, such as at least 2 further impurities, at least 3 further impurities, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 15, or at least 20 further impurities. Herein, for example, each of these further impurities can be independently selected, such that these further impurities can comprise at least one elemental impurity and / or at least one inorganic compound and / or at least one organic compound and / or at least one endotoxin and / or at least one bioburden.

[0031] To determine the at least one further total impurity level of the at least one further impurity, the respective impurity level of the at least one further impurity in each of the plurality of raw materials has to be considered. The method according to the present disclosure can comprise, for example, obtaining the respective impurity level of the at least one further impurity in each of the plurality of raw materials. By determining the at least one further total impurity level of the at least one further impurity in addition to the at least one total impurity level of the at least one impurity, a plurality of impurities can be considered for the evaluation of the raw material selection. This can encompass, for example, the case that in one of the plurality of raw materials the impurity level of the at least one impurity is comparably high, while the impurity level of the at least one further impurity is comparably low. In the case that the at least one total impurity level of the at least one impurity and the at least one total impurity level of the at least one further impurity can be subject to different criteria, such as different upper and / or lower limits, the method according to the present disclosure can be used to identify a raw material selection that fulfills all of these different criteria.

[0032] For example, the method according to the present disclosure has the advantage that the impurity levels of a plurality of impurities, such as at least one toxic impurity and at least one further toxic impurity and / or at least one functional impurity and / or at least one endotoxin and / or at least one bioburden and / or at least one inorganic compound and / or at least one organic compound, can be determined in a single optimization step.

[0033] According to exemplary embodiments, the method according to the present disclosure further comprises: - determining the at least one impurity level of the respective impurity level of the at least one impurity in each of the plurality of raw materials; and / or - outputting information indicative of the at least one total impurity level of the at least one impurity in the cell culture medium and / or the at least one further total impurity level of the at least one further impurity in the cell culture medium.

[0034] For example, determining the at least one impurity level of the respective impurity level of the at least one impurity in each of the plurality of raw materials can be understood as the device performing the method instructing the determination device to determine and subsequently transmit the respective impurity level by sending an electrical signal, thereby enabling the determination of the at least one impurity level.

[0035] The information indicative of the total impurity levels is output, e.g., can be understood as the device performing the method providing information, parameters and / or data indicative of the total impurity levels and / or at least one other total impurity level, e.g., via a communication interface of the device, e.g., in a suitable electronic format, such as a text format, a spreadsheet format or a database format. In one non-limiting example, a user interface can be used to output the total impurity levels to a user, e.g., in the form of a visual user output on a screen.

[0036] According to exemplary embodiments of the method of the present disclosure, the respective impurity levels of each raw material refer to respective batches of the raw material.

[0037] For example, a batch of a raw material is to be understood as a specific quantity or batch of the raw material produced or manufactured under a set of identical conditions or procedures, including the specific recipe, processing steps, equipment and environmental conditions required for manufacturing the raw material. For example, in a single batch of a raw material, the impurity levels of one or more impurities in the raw material are considered to be constant. When, for example, comparing the impurity levels of a specific impurity in several batches of the raw material, the impurity levels of the specific impurity can vary among the several batches of the raw material. For example, the respective batches of raw materials can be from different suppliers or from the same supplier.

[0038] According to exemplary embodiments, the method of the present disclosure further comprises: - determining, based at least in part on the total impurity levels, the recipe and the respective impurity levels of the cell culture medium, a respective contribution of one or more of the plurality of raw materials to the total impurity levels and / or at least one other total impurity level of the cell culture medium.

[0039] For example, the contribution of a raw material to a total impurity level of an impurity can be a ratio of (i) the impurity level of the impurity in the raw material, (ii) the total impurity level of the impurity in the cell culture medium, and further taking into account the proportion of the raw material in the cell culture medium. For example, such a contribution can be expressed in percentage, wherein the sum of the respective contributions of all raw materials to the same impurity in the cell culture medium can amount to 100%.

[0040] For example, a first raw material can contribute more to a total impurity level compared to a second raw material (e.g., because the impurity level of the impurity in the first raw material is higher than in the second raw material, and / or because the amount of the first raw material comprised in the cell culture medium is higher than the second raw material). In such an example, the first raw material can be referred to as a sensitive raw material.

[0041] For example, considering the respective total impurity levels of a plurality of impurities in a cell culture medium comprising a plurality of raw materials, a matrix representation, such as a sensitivity matrix, can provide the contribution of each raw material to each total impurity level.

[0042] Advantageously, the method of the present disclosure can rank the raw materials according to their contribution to the total impurity level, thereby improving the ability of the method to effectively determine those raw materials that have a major contribution to a certain total impurity level in a cell culture medium.

[0043] According to exemplary embodiments, the method of the present disclosure further comprises: - outputting information indicative of one or more raw materials having a contribution to the total impurity level and / or to at least one other total impurity level that is greater than a predefined threshold.

[0044] Outputting information indicative of the total impurity level, for example, can be understood as the device performing the method providing information, parameters and / or data representing the one or more raw materials, e.g. in an appropriate electronic format such as a text format, a spreadsheet format or a database format, e.g. via a communication interface of the device. In one non-limiting example, a user interface can be used to output the one or more raw materials to a user, e.g. by presenting the one or more raw materials in the form of a visual user output on a screen.

[0045] For example, the information indicative of one or more raw materials having a contribution to the total impurity level and / or to at least one other total impurity level that is greater than a predefined threshold can be used to generate a short list of sensitive raw materials from a plurality of raw materials that have a particularly strong influence on the total impurity level in a cell culture medium. Advantageously, only these sensitive raw materials can be used when further evaluating, e.g. optimizing, the total impurity level of a cell culture medium. For example, any further analysis, such as determining an allowed range or determining a respective preferred batch, can be based on only these sensitive raw materials, which can make the analysis, e.g. the corresponding optimization algorithm, more efficient.

[0046] According to exemplary embodiments, the method of the present disclosure further comprises: - determining whether the total impurity level and / or at least one other total impurity level meets at least one predefined criterion.

[0047] For example, a total impurity level of a certain impurity in a cell culture medium can meet a predefined criterion for the cell culture medium by being below or above a limit value for the total impurity level of that impurity in the cell culture medium. Considering the case of a plurality of total impurity levels in the cell culture medium, each total impurity level can be subject to a respective different predefined criterion. For example, the criterion can be predefined in accordance with prevailing regulations regarding physical, chemical, biological or microbiological properties of the cell culture medium, wherein these regulations and corresponding criteria have to be met in order to ensure the safety, efficacy and quality of the cell culture medium or of any product prepared at least partially based on the cell culture medium.

[0048] According to exemplary embodiments of the method of the present disclosure, determining whether the total impurity level and / or the at least one further total impurity level meets at least one predefined criterion further comprises at least one of: - determining whether the total impurity level and / or the at least one further total impurity level of the cell culture medium is below a predefined upper limit; and / or - determining whether the total impurity level and / or the at least one further total impurity level of the cell culture medium is above a predefined lower limit.

[0049] For example, the upper limit is to be understood as a predetermined maximum amount or maximum concentration of an impurity in the cell culture medium, above which an adverse effect on the cell cultivation and / or the cell culture product can be expected. For example, the total impurity level of any impurity in the cell culture medium should be avoided to exceed the respective upper limit of said impurity in the cell culture medium. For example, the upper limit can be based on the impurity and / or the cell culture medium. For example, each impurity in the cell culture medium can have an individual upper limit. Further, the upper limit of an impurity can be different or identical for different cell culture media.

[0050] For example, the lower limit is to be understood as a predetermined minimum amount or minimum concentration of an impurity in the cell culture medium, below which an adverse effect on the cell cultivation and / or the cell culture product can be expected. Typically, a lower limit is to be set for functional impurities, wherein the total impurity level of any functional impurity in the cell culture medium should be avoided to be below the respective lower limit. For example, the lower limit can be based on the impurity and / or the cell culture medium. For example, each impurity in the cell culture medium can have an individual lower limit. Further, the lower limit of an impurity can be different or identical for different cell culture media. Advantageously, the method of the present disclosure can set individually tailored limits for different cell culture media and / or for different impurities. This may, for example, encompass the case that the total impurity level of an undesired impurity, such as a toxic impurity, should be below an upper limit, while on the other hand the total impurity level of a desired impurity, such as a functional impurity, should be above a lower limit.

[0051] According to exemplary embodiments of the method of the present disclosure, determining whether the total impurity level and / or the at least one further total impurity level meets at least one predefined criterion is based at least in part on an upstream process model and / or a downstream process model of the cell culture medium.

[0052] For example, the upstream process model is to be understood as a modelling or calculation of the impurity level in the cell culture medium and / or the cell culture product based on an upstream process. Herein, for example, the upstream process is to be understood as the relevant processes prior to the production of the cell culture product by the cells in the cell culture medium as well as said production process. For example, the upstream process can comprise activities such as cell cultivation and / or comprise the addition of different cell culture media, such as feed media, over time and / or the partial removal of the cell culture medium, such as by at least one sampling step, and / or the addition of further substances during the cell cultivation, such as for pH adjustment, for preventing foam, etc.

[0053] For example, a downstream process model is to be understood as a modeling or calculation of the impurity levels in the cell culture product based on a downstream process. Herein, for example, a downstream process is to be understood as a process that purifies the cell culture product and prepares it for subsequent use. For example, a downstream process can include activities such as cell harvesting, cell disruption, chromatographic separation, filtration, and / or formulation (e.g., by incorporating such activities as a series of unit operations therein), etc.

[0054] Advantageously, by taking into account the upstream and / or downstream process model, the method of the present disclosure can more accurately estimate the total impurity level of at least one impurity in the cell culture medium and / or the cell culture product produced therefrom, which in turn can more accurately predict whether the total impurity level is likely to meet the respective predefined criteria, and whether the cell culture product can comply with regulatory requirements such as critical quality attributes (CQA). In some examples, the additional consideration of the upstream and / or downstream process can provide a more relaxed limit for the total impurity level in the cell culture medium, which can influence raw material selection.

[0055] According to an exemplary embodiment of the method of the present disclosure, the upstream process model includes an impurity level of at least one impurity in at least one supplement added to the cell culture medium in the upstream process, and the impurity level of the at least one impurity in the at least one supplement is added to the total impurity level before determining whether the total impurity level meets the at least one predefined criterion.

[0056] The upstream process model includes an impurity level of at least one impurity in at least one supplement, which may, for example, be understood as the upstream process model taking into account, considering, or encompassing the impurity level of the last impurity in the at least one supplement. For example, applying the upstream process model can include the step of adding the impurity level of the at least one impurity in the at least one supplement to the total impurity level before determining whether the total impurity level meets the at least one predefined criterion.

[0057] A supplement may, for example, be understood as an additive (e.g., a feed medium) that enhances the cell culture medium by providing essential nutrients, growth factors, or other ingredients necessary for cell growth and function optimization. In the upstream process, such a supplement (and, for example, other supplements) can be added to the cell culture medium, thereby forming a mixture of various media (e.g., a mixture of the cell culture medium and the feed medium). For example, the supplement added to the cell culture medium can contain at least one impurity present in the cell culture medium, such that the impurity level of the at least one impurity in the supplement can be added to the total impurity level of the at least one impurity in the cell culture medium.

[0058] To add the impurity level of the at least one impurity in the at least one supplement to the total impurity level of the at least one impurity in the cell culture medium, for example, the numerical value of the impurity level can be added to the numerical value of the total impurity level. Additionally or alternatively, the impurity level and the total impurity level can be added according to a mass balance approach, which for example indicates the accumulation of the at least one impurity in the upstream process.

[0059] The impurity level of the at least one impurity in the at least one supplement is added to the total impurity level before determining whether the total impurity level meets the at least one predefined criterion. This can for example be understood to mean that the impurity level is added to the total impurity level before determining whether this additive result (for example, as a new total impurity level taking into account the downstream process) meets the at least one predefined criterion. For example, the method according to the present disclosure can comprise the step of adding the value of the impurity level of the at least one impurity in the at least one supplement to the value of the total impurity level in the cell culture medium before determining whether the total impurity level meets the at least one predefined criterion.

[0060] According to an exemplary embodiment of the method according to the present disclosure, the downstream process model comprises at least one factor representing a change in the total impurity level in the downstream process, and it is determined, based at least in part on the at least one factor, whether the total impurity level meets the at least one predefined criterion.

[0061] The downstream process model comprises at least one factor representing a change in the total impurity level, which can for example be understood to mean that the downstream process model takes into account, considers or encompasses at least one factor representing a change in the total impurity level. For example, the downstream process of the cell culture medium can comprise one or more downstream process steps, wherein the total impurity level of the at least one impurity in the cell culture medium changes in each process step of the downstream process. Each process step can be represented by a factor representing a change in the total impurity level in the respective process step.

[0062] For example, a change in the total impurity level in the cell culture medium in the downstream process can be understood as a change (such as a decrease or an increase) in the total impurity level in the cell culture medium during the downstream process of the cell culture medium. This change in the total impurity level can for example be caused by dilution, filtration, purification (such as protein A chromatography) and / or similar procedures, which affect the at least one impurity in the cell culture medium.

[0063] For example, the at least one factor representing a change in the total impurity level in the downstream process can be given by a multiplication factor, wherein the total impurity level is multiplied by the at least one multiplication factor. Thereafter, for example, it can be determined whether the result of this multiplication (for example, as a new total impurity level taking into account the downstream process) meets the at least one predefined criterion. For example, the method according to the present disclosure can comprise the step of multiplying the value of the total impurity level by the value of the at least one factor before determining whether the total impurity level meets the at least one predefined criterion.

[0064] According to exemplary embodiments, the method of the present disclosure further comprises: - outputting information indicating that the plurality of raw materials and the recipe are suitable for use in manufacturing the cell culture medium, if it is determined that the total impurity level and / or the further total impurity level fulfills at least one predefined criterion.

[0065] Outputting information indicating that the plurality of raw materials and the recipe are suitable for use in manufacturing the cell culture medium, for example, can be understood to mean that the device performing the method can provide information, parameters and / or data representing that the plurality of raw materials is suitable for use in manufacturing the cell culture medium, e.g. in an appropriate electronic format such as a text format, a spreadsheet format or a database format, e.g. via a communication interface of the device. In one non-limiting example, a user interface can be used to output one or more raw materials to a user, e.g. by presenting one or more raw materials in the form of a visual user output on a screen.

[0066] In another example, if it is determined that the total impurity level and / or the further total impurity level fulfills at least one predefined criterion, the method can further comprise manufacturing the cell culture medium using the plurality of raw materials and the recipe indicated to be suitable, e.g. by instructing or controlling a manufacturing device.

[0067] According to exemplary embodiments, the method of the present disclosure further comprises: determining an allowed range of impurity levels of at least one impurity in at least one raw material of the plurality of raw materials based at least in part on the recipe, the respective impurity level of the at least one impurity in each raw material of the plurality of raw materials, and the at least one predefined criterion.

[0068] For example, it can be determined that the impurity level of the at least one impurity of the at least one raw material can vary within the allowed range while the total impurity level of the first impurity in the cell culture medium still fulfills the predefined criterion (e.g. the upper limit and / or the lower limit). In other words, the total impurity level of the at least one impurity in the cell culture medium determined based at least in part on the recipe and any impurity level of the at least one impurity of the at least one raw material within the determined allowed range fulfills the at least one predefined criterion. For example, in case the impurity level in the raw material varies within the allowed range, the at least one predefined criterion is still fulfilled with a certain probability. Herein, the allowed range of impurity levels of a certain impurity can be determined to ensure that the probability that the total impurity level of the impurity does not fulfill the predefined criterion is below a threshold probability.

[0069] According to exemplary embodiments of the method of the present disclosure, the allowed range of impurity levels of the at least one impurity in at least one raw material of the plurality of raw materials is determined based on the respective impurity level of the at least one impurity in those raw materials which are determined to contribute more than a predefined threshold to the total impurity level of the at least one impurity in the cell culture medium.

[0070] For example, determining the allowed range of impurity levels of a particular impurity can be based not on the respective impurity levels of the particular impurity in each of a plurality of raw materials for the cell culture medium, but on the respective impurity levels of the particular impurity in those raw materials for which it has been determined that their contribution to the total impurity level of this particular impurity is greater than a predefined threshold. In this way, the efficiency of an optimization algorithm, which can be used, for example, for determining the allowed range, can be improved.

[0071] The allowed range of impurity levels in a raw material can be given, for example, by upper and lower limits of the impurity level, wherein the upper and lower limits can be understood as allowed limits of the impurity level.

[0072] Advantageously, the method according to the present disclosure provides an allowed range of impurity levels for a particular raw material, which ensures that a predetermined criterion of the cell culture medium is met (e.g., with a certain probability) as long as the impurity level in the raw material varies within the allowed range. Considering, for example, that the impurity level in a raw material can vary from batch to batch of the raw material, the allowed range advantageously indicates whether a batch of the raw material can still be used for manufacturing the cell culture medium or whether it should be discarded.

[0073] According to one exemplary embodiment, determining the allowed range of the respective impurity level of at least one impurity of at least one raw material of the plurality of raw materials further comprises: - calculating a probability that the total impurity level of the at least one impurity in the cell culture medium does not meet the at least one predefined criterion based at least partly on the initial range of the impurity level of the at least one impurity of the at least one raw material; and - determining the allowed range of the respective impurity level based on the calculated probability and the initial range of the impurity level, in particular by using an optimization algorithm.

[0074] For example, the initial range of the impurity level of a particular impurity in a raw material can be determined based on the statistical variation of the impurity level in a plurality of batches of the raw material. Such an initial range can be retrieved, for example, from a database of raw materials.

[0075] Calculating a probability that the total impurity level of the at least one impurity in the cell culture medium does not meet or violate the at least one predefined criterion can comprise, for example, selecting a plurality of impurity levels (e.g., as samples) of the at least one impurity within the initial range of the impurity level of the at least one impurity of the at least one raw material. Subsequently, it can be determined how many of the selected impurity levels within these initial ranges are likely to violate the at least one predefined criterion. For example, the proportion of impurity levels that violate the predefined criterion out of all selected impurity levels within the allowed range can be indicative of the probability of violating the at least one predefined criterion.

[0076] Subsequently, the probability of violating the at least one predefined criterion can be compared to a threshold probability, which for example indicates whether a certain violation probability is still acceptable. If the violation probability is higher than the threshold probability, the initial range can be narrowed to reach the allowed range. If the violation probability is lower than the threshold probability, the initial range can be expanded to reach the allowed range.

[0077] In a further example, determining the allowed range based on the initial range can comprise executing an optimization algorithm aiming at minimizing an objective function comprising the difference between the violation probability and the threshold probability. For example, such an optimization algorithm can be performed in multiple iterations, wherein the allowed range as output of one iteration can be used as input for the next iteration of the optimization algorithm (e.g. as initial range).

[0078] For example, the initial range of the impurity level of the at least one impurity can be given by a mean value and a standard deviation of the mean value. Then, determining the allowed range for the impurity level of each impurity based on the calculated probability and the initial range of the impurity level can comprise determining the mean value and / or the standard deviation of the initial range.

[0079] According to exemplary embodiments, the method of the present disclosure further comprises: outputting information indicative of the allowed range for the impurity level of each of the at least one raw material.

[0080] Outputting the information indicative of the allowed range for example can be understood as meaning that the device performing the method can provide information, parameters and / or data indicative of the allowed range, e.g. in an appropriate electronic format such as a text format, a spreadsheet format or a database format, e.g. via a communication interface of the device. In one non-limiting example, a user interface can be used to output one or more raw materials to a user, e.g. by presenting one or more raw materials in the form of a visual user output on a screen.

[0081] According to exemplary embodiments, the method of the present disclosure further comprises: determining a deviation of the total impurity level of the cell culture medium and / or of the at least one further total impurity level from a respective reference total impurity level in the cell culture medium.

[0082] For example, the deviation of a total impurity level of a particular impurity is to be understood as the amount or degree of difference or deviation between the total impurity level and the reference total impurity level of the particular impurity in the cell culture medium. Herein, for example, the reference total impurity level can refer to the allowed range and / or the upper limit and / or the lower limit of the respective impurity in the cell culture medium. For example, if it is determined that the total impurity level of the cell culture medium and / or the at least one further total impurity level is within the allowed range and / or below the upper limit, optionally above the lower limit, the deviation can preferably be determined as zero. Advantageously, the method according to the present disclosure can allow for a precise customization to the requirements of the cell culture process to be optimized.

[0083] For example, the objective function can represent the deviation of the total impurity level from the respective reference total impurity level. Such an objective function can additionally represent the deviation of the at least one other total impurity level from the respective at least one other reference total impurity level. In this example, minimizing the objective function can advantageously take into account potential deviations of multiple total impurity levels from respective reference total impurity levels.

[0084] According to exemplary embodiments, the method of the present disclosure further comprises: - determining, based at least in part on the total impurity level of the cell culture medium and / or the respective impurity levels of the at least one other total impurity, the formulation, and each of the raw materials, a respective contribution of one or more of the raw materials to the determined deviation; and - determining one or more of the raw materials having a contribution to the determined deviation greater than a predefined threshold.

[0085] For example, the contribution of a raw material to the determined deviation between the total impurity level in the cell culture medium and the respective reference total impurity level can be given by the contribution of the raw material to the total impurity level of the determined deviation. Herein, the contribution to the total impurity level can be determined, for example, according to the method described above.

[0086] Advantageously, the method of the present disclosure can rank the raw materials according to the extent of their contribution to the deviation of the total impurity level from the respective reference total impurity level, which can improve the ability of the method to effectively determine those raw materials that need to be optimized to ensure, for example, that the total impurity level meets the predefined criteria.

[0087] For example, by determining one or more of the raw materials having a contribution to the determined deviation greater than a predefined threshold, a list of problem causes can be identified among the plurality of raw materials that have a particularly strong influence on the deviation of the total impurity level in the cell culture medium.

[0088] According to exemplary embodiments, the method of the present disclosure further comprises: - determining a respective preferred batch of at least one of the plurality of raw materials, wherein the deviation of the total impurity level of the cell culture medium and / or the at least one other total impurity level from the respective reference total impurity level of the cell culture medium is reduced, in particular minimized, by using an optimization algorithm based at least in part on the impurity level of at least one impurity of the preferred batch of the at least one raw material.

[0089] For example, considering a plurality of raw materials comprised in a cell culture medium, each raw material can refer to a respective batch. In this example, reducing the deviation of the total impurity level in the cell culture medium from the respective reference total impurity level by determining a preferred batch for at least one of the raw materials can be understood as meaning that for this at least one raw material, an alternative batch is determined, based on which the deviation of the total impurity level for a particular impurity is reduced (e.g. minimized). For example, based on the determined deviation of the total impurity level for a particular impurity, the impurity level for this particular impurity in the alternative raw material batch can differ (e.g. be higher or lower) from the impurity level for this particular impurity in the original raw material batch. In this case, the alternative batch can be referred to as the preferred batch. This difference in the impurity level between the original batch and the preferred batch can change the total impurity level in the cell culture medium in a way that reduces the deviation of the total impurity level.

[0090] According to exemplary embodiments, a raw material equivalent to at least one raw material (e.g. referred to as an equivalent raw material) can be determined (e.g. as an alternative to the preferred batch of the at least one raw material), wherein based at least in part on the impurity level of at least one impurity in the equivalent raw material, the deviation of the total impurity level and / or at least one other total impurity level in the cell culture medium from the respective reference total impurity level in the cell culture medium is reduced.

[0091] In this example, the method of the present disclosure further comprises: - determining a respective equivalent raw material for at least one of the plurality of raw materials, wherein based at least in part on the impurity level of at least one impurity in the respective equivalent raw material, the deviation of the total impurity level and / or at least one other total impurity level in the cell culture medium from the respective reference total impurity level in the cell culture medium is reduced, in particular by minimizing the deviation of the total impurity level and / or at least one other total impurity level from the respective reference total impurity level using an optimization algorithm.

[0092] For example, the equivalent raw material of the at least one raw material can be chemically and biologically equivalent to the at least one raw material, which can be understood to mean that replacing the at least one raw material with the equivalent raw material in the cell culture medium does not result in significant side effects. For example, in the equivalent raw material, the degree of hydration can be different (e.g. by being higher or lower in solvent content), or for example, counterions can be exchanged if the exchange of counterions has no adverse effect on cell culture and / or cell culture products, wherein for example the counterions do not significantly change the content of the respective elements in the composition, or preferably the exchange is accompanied by a corresponding change in another raw material, so that the ion content in the cell culture medium is essentially the same as the ion content of the cell culture medium without the replacement of the raw material. This can have the advantage that there are more essentially identical raw materials to choose from, while there are also more suitable impurity profiles to choose from.

[0093] For example, minimizing the deviation from the respective reference total impurity level can be understood as meaning minimizing the deviation within a certain range around the reference total impurity level.

[0094] For example, the deviation between the determined total impurity level and the reference total impurity level can be expressed using an objective function. Then, this deviation can be reduced by finding the minimum of the objective function. In one example, the preferred batches of raw materials can be determined by calculating the objective function for each possible combination of batches of the plurality of raw materials. Then, the preferred batch for a particular raw material can be determined from the combination of batches that leads to the minimum of the objective function. To improve the efficiency of this approach, if a large number of raw materials in the cell culture medium (e.g., 50 raw materials) are evaluated, it is not necessary to consider each of the plurality of raw materials, but only those raw materials whose contribution to the respective total impurity level is greater than a predefined threshold. Another way to improve the efficiency when determining the preferred batches can be to use a global optimizer to minimize the objective function and determine the optimal combination of batches. This approach can for example involve a transformation function for solving a discrete problem (e.g., selecting the preferred batches), while minimizing the objective function can require solving a continuous optimization problem. Advantageously, the method of the present disclosure can allow selecting batches of raw materials such that the resulting cell culture medium and cell culture product comply with standards such as the ICH guidelines, and / or with the CQA of a given cell line, cell culture medium and / or process.

[0095] According to an exemplary embodiment of the method of the present disclosure, the respective preferred batch of one or more raw materials is determined for which the contribution to the determined deviation is greater than a predefined threshold.

[0096] Considering the example where preferred batches of several raw materials of the plurality of raw materials are determined that reduce the deviation of the total impurity level, for those raw materials that have been determined to contribute to the determined deviation greater than a predefined threshold as sensitive raw materials, only the respective preferred batch can be determined for them. Advantageously, replacing the original batch of those sensitive raw materials with the respective preferred batch that reduces the deviation can be sufficient to make them meet the predetermined standards. In such an example, the efficiency of the optimization algorithm that determines the preferred batches can be improved, as it can skip raw materials that contribute to the deviation to be reduced to a rather low extent.

[0097] According to an exemplary embodiment of the method of the present disclosure, a plurality of recipes for each cell culture medium is obtained, wherein each recipe of the plurality of recipes shows a proportion of a plurality of raw materials in the respective cell culture medium; and the method further comprises at least one of: - determining a respective allowed range of impurity levels of at least one impurity based on the plurality of recipes; and / or - determining a respective preferred batch of at least one raw material based on the plurality of recipes.

[0098] For example, the plurality of cell culture media can be understood as a portfolio of cell culture media, which can be prepared according to respective recipes of a plurality of recipes, which can show the proportions of the plurality of raw materials in the respective cell culture media. While the recipes and the plurality of raw materials can differ from each other for the respective cell culture media of the portfolio, the respective cell culture media can for example share at least one or more common raw materials. In such an example, the respective allowed ranges of impurities and / or the respective preferred batches of the common raw materials can be determined. In this case, the determination of the respective allowed ranges and / or the determination of the respective preferred batches can be dependent on the respective recipe and further dependent on the respective raw material of the plurality of raw materials of the cell culture media of the portfolio. Advantageously, when optimizing the entire portfolio of cell culture media, the method according to the present disclosure can for example ensure that all media of the portfolio meet the predefined criteria. This approach can ensure that as few raw materials as possible are assigned to specification limits, which can add additional workload to the production procurement process.

[0099] According to an exemplary embodiment of the method of the present disclosure, the at least one impurity and / or the at least one further impurity is one of: a toxic impurity; or a functional impurity; or an organic compound; or an inorganic compound; or a biological load; or an endotoxin.

[0100] For example, consider a single impurity, which can be a toxic impurity, a functional impurity, an organic compound, an inorganic compound, a biological load, or an endotoxin. In another example, consider one impurity (i.e., a first impurity) and another impurity (i.e., a second impurity), the first impurity can be a toxic impurity, a functional impurity, an organic compound, an inorganic compound, a biological load, or an endotoxin, and the second impurity can be a toxic impurity, a functional impurity, an organic compound, an inorganic compound, a biological load, or an endotoxin. In another example, consider yet another impurity (i.e., a third impurity), the third impurity can be a toxic impurity, a functional impurity, an organic compound, an inorganic compound, a biological load, or an endotoxin, and the second impurity can be a toxic impurity, a functional impurity, an organic compound, an inorganic compound, a biological load, or an endotoxin. In another example, the at least one further impurity is understood to include any number of further impurities, for example at least 2 further impurities, at least 3 further impurities, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 15, or at least 20 further impurities.

[0101] wherein each - indented item can refer to a class of impurities, as described in more detail above.

[0102] Advantageously, the method of the present disclosure can be used, for example, to even increase the impurity levels in the cell culture medium. For example, in case of functional impurities, such a measure can be particularly desirable, different from the regular use of the term "impurity", functional impurities can be required in the cell culture medium to ensure proper cell cultivation. Alternatively or additionally, such a measure can also be desirable when increasing the impurity levels within the allowed range and / or increasing the impurity levels to a level below the upper limit for each impurity. Alternatively or additionally, such a measure can also be desirable when decreasing the impurity level of one impurity by increasing the impurity level of another impurity.

[0103] According to an exemplary embodiment of the method of the present disclosure, it can be determined that a particular functional impurity needs to be added to the cell culture medium. This can be understood, for example, as adding the functional impurity to the cell culture medium separately (e.g., independent of the addition of the raw materials in the cell culture medium). For example, by determining that the total impurity level of the functional impurity in the cell culture medium is below a predefined lower limit in the cell culture medium, it can be determined that the total impurity level of the functional impurity in the cell culture medium does not meet at least one predefined criterion. In such an example, it can be determined that the functional impurity (e.g., a particular amount thereof) needs to be added to the cell culture medium in order to meet the predefined criterion by adding the functional impurity to the cell culture medium.

[0104] According to the present exemplary embodiment, the method of the present disclosure further comprises: - determining that the functional impurity needs to be added to the cell culture medium if it is determined that the total impurity level of the functional impurity in the cell culture medium does not meet at least one predefined criterion; in particular, - outputting information indicating that the functional impurity needs to be added to the cell culture medium.

[0105] Advantageously, the method of the present disclosure can optimize the total impurity level even in case the stock does not have raw materials with a suitable impurity profile.

[0106] According to an exemplary embodiment of the method of the present disclosure, the cell culture medium is in liquid form, and wherein the formulation of the cell culture medium further indicates a hydration procedure for the plurality of raw materials.

[0107] For example, the hydration procedure is to be understood as a process for preparing the liquid cell culture medium based at least in part on the solid cell culture medium. Such a process can comprise, for example, steps such as adding at least one solvent (e.g., water), stirring, and sterile filtration. Depending on the cell culture medium, the hydration procedure can also comprise, for example, other steps such as adjusting the pH value and / or adding other substances.

[0108] With further consideration of the above-disclosed features, the disclosure of the method steps of the methods of the disclosure should also be considered as a disclosure of apparatus configured to perform the respective method steps, and as a disclosure of apparatus comprising means for performing the respective method steps. Likewise, the disclosure of apparatus configured to perform the method steps or of apparatus comprising means for performing the method steps should also be considered as a disclosure of the method steps themselves.

[0109] It is to be understood that the embodiments disclosed herein are merely examples and not limiting.

[0110] Other features of the present disclosure will become apparent from consideration of the following detailed description taken in conjunction with the accompanying drawings. It is understood that the drawings merely show possible embodiments, and that it is not limiting the scope of the disclosure to these exact drawings. It is also understood that the drawings are not drawn to scale, and are only intended to conceptually illustrate the structures and procedures described. BRIEF DESCRIPTION OF DRAWINGS

[0111] Some example embodiments of the present disclosure will now be described with reference to the accompanying drawings.

[0112] Figure 1 An example environment in which methods according to the present disclosure can be performed is shown; Figure 2a , 2b Examples of various starting materials for cell culture media according to the present disclosure are shown; Figure 3 A flowchart illustrating one example embodiment of a method according to the present disclosure is shown; Figure 4 A flowchart illustrating another example embodiment of a method according to the present disclosure is shown; Figure 5 A flowchart illustrating another example embodiment of a method according to the present disclosure is shown; Figure 6 A flowchart illustrating another example embodiment of a method according to the present disclosure is shown; Figures 7a to 7c Another example embodiment of a method according to the present disclosure is shown; Figure 8 A flowchart illustrating another example embodiment of a method according to the present disclosure is shown; Figure 9 A scheme illustrating another example embodiment of a method according to the present disclosure is shown; Figure 10a , 10b Example experimental data according to the present disclosure is shown; Figure 11 Further example experimental data according to the present disclosure is shown; Figures 12a to 12d Further exemplary experimental data according to the present disclosure is shown; Figures 13a to 13d Further exemplary experimental data according to the present disclosure is shown; Figure 14 Further exemplary experimental data according to the present disclosure is shown; Figure 15 Further exemplary experimental data according to the present disclosure is shown; Figure 16 A schematic diagram showing one exemplary downstream process according to the present disclosure is shown; Figure 17 Further exemplary experimental data according to the present disclosure is shown; Figure 18 Further exemplary experimental data according to the present disclosure is shown; Figure 19 Further exemplary experimental data according to the present disclosure is shown; Figure 20 Further exemplary experimental data according to the present disclosure is shown; Figure 21 A schematic diagram showing an exemplary embodiment of an apparatus according to the present disclosure is shown; and Figure 22 An exemplary embodiment of a storage medium is shown. DETAILED DESCRIPTION

[0113] The following description is intended to further enhance the understanding of the present disclosure and should be read in conjunction with the description of exemplary embodiments of the present disclosure as described in the above sections of the specification.

[0114] Figure 1 An exemplary environment 100 in which methods according to the present disclosure can be performed is shown.

[0115] As a general example, a cell culture medium can comprise a plurality of different substances, such as amino acids, amino acid derivatives, nucleotides, salts, biological amines, fats, sugars, vitamins, dyes or complex biological additives, each of which is present in a predetermined amount. Herein, each of these substances can be purchased individually as a raw material as referred to herein. The cell culture medium can be made in a powdered form, which needs to be hydrated by the customer before use. Alternatively, the cell culture medium can also be prepared, for example, in a liquid form.

[0116] Without being limited thereto, the cell culture medium can contain at least one of the following raw materials: biotin, calcium chloride (CaCl2, anhydrous), choline chloride, copper sulfate (CuSO4 5 H2O), calcium D-pantothenate, D-glucose (dextrose), iron nitrate (Fe(NO3)3 9 H2O), iron sulfate (FeS04 7 H2O), folic acid, glycine, sodium hypoxanthine, i-inositol, L-alanine, L-arginine hydrochloride, L-asparagine H2O, L-aspartic acid, L-cysteine hydrochloride H2O, L-cystine 2 HCI, L-glutamic acid, L-glutamine, L-histidine hydrochloride H2O, linoleic acid, lipoic acid, L-isoleucine, L-leucine, L-lysine hydrochloride, L-methionine, L-phenylalanine, L-proline, L-serine, L-threonine, L-tryptophan, L-tyrosine disodium salt dihydrate, L-valine, magnesium sulfate (MgS04, anhydrous), nicotinamide, phenol red, potassium chloride (KC1), putrescine 2 HCI, pyridoxine hydrochloride, riboflavin, sodium bicarbonate (NaHC03), sodium chloride (NaCl), sodium phosphate dibasic (Na2HP04, anhydrous), sodium phosphate monobasic (NaH2P04 H2O), pyruvic acid sodium, thiamine hydrochloride, thymidine, vitamin B12 and / or zinc sulfate (ZnS04 7 H2O).

[0117] Without being limited thereto, the cell culture medium can contain at least one of the following raw materials: biotin, calcium nitrate (Ca(N03)2 4 H2O), choline chloride, D-calcium pantothenate, D-glucose (dextrose), folic acid, glutathione (reduced), glycine, i-inositol, L-arginine, L-asparagine, L-aspartic acid, L-cystine 2 HCI, L-glutamic acid, L-glutamine, L-histidine, L-hydroxyproline, L-isoleucine, L-leucine, L-lysine hydrochloride, L-methionine, L-phenylalanine, L-proline, L-serine, L-threonine, L-tryptophan, L-tyrosine disodium salt dihydrate, L-valine, magnesium sulfate (MgS04, anhydrous), nicotinamide, p-aminobenzoic acid, phenol red, potassium chloride (KC1), pyridoxine hydrochloride, riboflavin, sodium bicarbonate (NaHC03), sodium chloride (NaCl), sodium phosphate dibasic (Na2HP04, anhydrous), thiamine hydrochloride and / or vitamin B12.

[0118] Other examples of raw materials for cell culture media are known in the art, for example, as evidenced in the prior patent applications WO 2007 / 036291 A2 and WO 2009 / 087087 A1, the entire contents of which are hereby incorporated by reference.

[0119] Due to the manufacturing process of the raw materials, each raw material can contain, in addition to the target substance, one or more impurities, such as one or more toxic substances (in particular elements and / or compounds), one or more functional substances (in particular elements and / or compounds), and / or biological load, and / or endotoxins. In some examples, impurities can be understood as not of animal origin. These impurities end up in the cell culture medium prepared from the respective raw materials. For example, an amount of a certain impurity in the cell culture medium exceeding a certain limit can lead to adverse effects, such as a reduced product yield, a compromised product quality, or even a contamination of the cell culture product, which can not be usable or acceptable in downstream applications, such as human therapeutic applications. Therefore, the manufacturer of the cell culture medium strives to use only those raw materials that avoid impurities and any such adverse effects.

[0120] For example, in the case of cell culture products such as biopharmaceuticals, there are strict regulations for their physical, chemical, biological or microbiological properties, which should be controlled within predefined limits to ensure the safety, efficacy and quality of the product. In the industry, these so-called critical quality attributes (CQA) are generally considered essential for the development, manufacturing and regulatory approval of cell culture products such as biopharmaceuticals. Cell culture media can, for example, affect the CQA of the product, such as purity, post-translational modifications, protein folding and disulfide bond formation, and / or the safety of the product when administered to a patient. In order to provide a pure, high-quality and safe cell culture product, the cell culture process must be carried out in a cell culture medium that does not introduce harmful contaminants and / or impurities to the cell culture or to the finished drug product itself.

[0121] For example, known guidelines such as the EMA scientific guideline ICH Q3D provide limits for certain elemental impurities in finished drug products. In addition to this, there can be non-binding estimates (e.g. literature-based estimates) that relate the limits of ICH Q3D to limits for elemental impurities in raw materials for cell culture media, taking into account dilution effects in downstream processing steps. These estimates can have the disadvantage of being imprecise, for example, because the limits derived from finished drug product regulations (such as ICH Q3D) are not specifically tailored for such use. Therefore, it can be challenging to identify one of the many raw materials that causes adverse effects in the cell culture medium. Furthermore, if rules from scientific guidelines, for example, related to raw materials rather than drug products, are applied, it can lead to a specific raw material or batch thereof being discarded, which can cause unnecessary waste and cost increases for the manufacturer of the cell culture medium and the end customer.

[0122] Due to the large number of raw materials in a given cell culture medium and the fact that there are usually multiple batches of each raw material at the manufacturing site, it can be challenging to calculate the final impurity levels for all possible batch combinations in a given cell culture medium.

[0123] Therefore, it is necessary to provide a reliable, rapid, and accurate method for calculating the amounts of various impurities in cell culture media. Preferably, the method according to this disclosure should be configured to detect raw materials that need to be changed or replaced, thereby improving (e.g., reducing or increasing) the amounts of various impurities in the cell culture media. More preferably, the method should allow the use of raw materials with impurity levels within permissible ranges, thereby achieving stability, flexibility, and economy in raw material procurement by allowing variations in impurity levels within permissible ranges. Also preferably, the method should be able to identify cell culture media or combinations of cell culture media that comply with regulatory and / or CQA requirements.

[0124] refer to Figure 1 Non-limiting examples, and according to the method of this disclosure, a cell culture medium to be optimized, comprising a variety of ( Raw materials 110, 120, 130 (see Raw Materials A, B, ..., n) are listed below. Formulation 140 (see “Culture Medium Ingredients Table”) is available in this document, showing the proportions of various raw materials in the cell culture medium (see reference below). Figure 2a and Figure 2b As shown, for example, raw material A accounts for 40%, raw material B accounts for 30%, etc. In addition, multiple other types can be obtained ( The level of each impurity in at least one impurity of each of the following raw materials: (types) Figure 1 In the example shown, three impurities are considered (see Figure 1 The "Trait 1", "Trait 2", and "Trait 3" in the text retrieve multiple (...) The levels of each of the three impurities in each of the raw materials (see) Figure 1 The values ​​of “Trait 1”, “Trait 2” and “Trait 3” in the dataset.

[0125] Further reference Figure 1 The example shown can be assumed to produce a product based on multiple ( Cell culture media containing raw materials 110, 120, 130 and corresponding formulation 140. In this document, the method disclosed includes at least a portion based on formulation 140 (see...). Figure 1 (See "Culture Material List") and the levels of each impurity (see...) Figure 1 The values ​​of "Trait 1", "Trait 2", and "Trait 3" in the data are used to determine the total impurity levels of each of the three impurities in the cell culture medium. According to the method of this disclosure, for example, the cell culture medium can be considered to be in powder form (see [link to disclosure]). Figure 1 The "powdered culture medium" in the text is still in liquid form (see [link]). Figure 1(Referring to "Liquid Culture Medium Production"). In liquid form, cell culture medium formulation 140 can indicate multiple (…). A hydration process for raw materials 110, 120, and 130, which includes, for example, the steps of adding at least one solvent (such as water) to a cell culture medium in powder form, stirring, and aseptic filtration.

[0126] like Figure 1 As shown, determining the total impurity level of at least one impurity in the cell culture medium, at least in part based on formulation 140 and the levels of each impurity, can be used to optimize the selection of raw materials (e.g., cell culture medium in powder or liquid form). Optimizing the selection of raw materials may include the steps described in the following examples.

[0127] In one example, multiple ( ) can be determined One or more of the following raw materials are used in the production of raw materials: Figure 1 The contribution of the total impurity level to the three impurities shown can, for example, include determining the sensitivity matrix, as will be referenced below. Figure 2a and 2b Further description. For example, based on the higher level of the first impurity in raw material A (see... Figure 1 By examining the "Trait 1" of raw material A and its content in the cell culture medium according to the corresponding formulation, it can be determined that raw material A contributes more to the total impurity level of the first impurity than other raw materials. For example, it can be inferred that the cell culture medium is particularly sensitive to the exact impurity level in raw material A, and therefore, when used for culture medium production, the exact impurity level in raw material A should be kept constant (e.g., constant within a predefined range).

[0128] In another example, it can be determined whether the total impurity level of three impurities in the cell culture medium meets at least one predefined criterion. For example, this could include determining whether a specific total impurity level in the cell culture medium is below a predefined upper limit (e.g., considering that the specific impurity level is associated with a toxic impurity), and / or determining whether a specific total impurity level in the cell culture medium is above a predefined lower limit (e.g., considering that the specific impurity level is associated with a functional impurity). In another example, it can be determined whether the deviation between a specific total impurity level in the cell culture medium and the respective reference total impurity levels in the cell culture medium is determined, wherein multiple ( Preferred batches of at least one of the raw materials can reduce this deviation, as detailed in Figure 7 below. Figure 8 As stated above.

[0129] In another example, multiple ( ) can be determined The permissible range for the level of a specific impurity in at least one of the raw materials (a, b, c) is defined. For example, the impurity level of the first impurity in raw material A can be determined (see [reference]).Figure 1 Trait 1” of raw material A) can vary within an allowable range while the total impurity level of the first impurity in the cell culture medium still satisfies the predefined criteria (e.g., upper and / or lower limits). More details regarding determining the allowable range will be referred to Figures 4 to 6 Description.

[0130] Further referring to the example shown in Figure 1 FIG. 1, the method according to the present disclosure can also take into account upstream process model 150 and downstream process model 160 of the cell culture medium. Thus, the optimization of raw material selection can not only be applicable to the production of the cell culture medium, but also as shown in Figure 1 FIG. 1, to the upstream process of the cell culture medium production and the subsequent downstream process. For example, the upstream process can refer to an early stage of the production process, where cells (e.g., mammalian cells) can be, for example, genetically modified to produce a desired protein (e.g., a therapeutic antibody). For example, this stage can typically include activities such as cell culture medium optimization, cell line development, genetic engineering, and / or cell bank establishment, etc. In another example, the downstream process can refer to a later stage of the production process, for example, where the produced protein can be purified and prepared for use. For example, this stage can typically include activities such as cell harvesting, chromatographic separation, filtration, and / or formulation, etc.

[0131] In a general example, the drug product 170 as Figure 1 shown in the process can be subject to regulatory agencies, and thus, the optimization of raw material selection can be improved not only by determining the total impurity level in the cell culture medium, but also by potential changes in such impurity levels that can be introduced during the upstream and downstream process. In some examples, the additional consideration of the upstream and downstream process can set more lenient limits on the total impurity level in the cell culture medium as well as the impurity level in the raw materials. For example, since certain impurity limits are not covered in the cell culture medium optimization or the upstream processing, the downstream process can be modeled to serve as an additional tool for making the raw material selection. More details regarding considering the upstream process model and the downstream process model will be referred to Figure 3 Description.

[0132] As Figure 1Exemplarily, the optimized raw material selection according to the method of the present disclosure can bring more advantages. For example, the most suitable raw material can be selected according to the given application and predefined criteria. Further, for example, key impurities which have a significant impact on the performance of the resulting cell culture medium can be determined. Setting an allowed range (e.g. by defining an allowed boundary) for these key impurities can more robustly procure raw materials and minimize the batch failure rate. In a further example, by setting an allowed range only for key impurities and / or only for raw materials with a larger impact, the proposed method can set more relaxed limits in terms of impurity content of raw material batches, which in turn can increase the robustness and flexibility of the supply chain, while also reducing costs due to a reduced need for special small batch raw materials.

[0133] Figure 2a and 2b Exemplary examples of various raw materials for a cell culture medium according to the present disclosure are shown.

[0134] In one example, the method according to the present disclosure can be used to optimize the raw material composition in a cell culture medium comprising two raw materials. For the two raw materials, which are referred to as raw material A and raw material B, two batches of each raw material are available. In the following, batch 1 of raw material A will be referred to as “A1”, batch 2 of raw material A will be referred to as “A2”, batch 1 of raw material B will be referred to as “B1”, and batch 2 of raw material B will be referred to as “B2”. Further, for each raw material, a reference batch related to a commercially available cell culture medium is available, which will be referred to as “Aref” and “Bref” in the following for raw materials A and B, respectively. The impurity levels in such reference batches can be determined, for example, by assay or estimation.

[0135] For each of the batches A1, A2, B1, B2, Aref, Bref, the impurity level of the functional impurity manganese (Mn) is obtained (e.g. by assay or by retrieving the impurity level from a database), as shown in the table of Figure 2a For the cell culture medium, a recipe is obtained which shows that the cell culture medium consists of 80% of raw material A and 20% of raw material B. From these exemplary data, the total Mn impurity level in the cell culture medium composed of raw materials from batches Aref and Bref is determined to be 1.32 ppm (= [0.8 1.6 ppm] + [0.2 0.2 ppm]), which can be referred to as the reference total impurity level of Mn in the cell culture medium. For batches A1, A2, B1, B2, the deviation of the total Mn impurity level from the reference total impurity level is determined, and in addition, it is determined whether the total Mn impurity level is below an upper limit.

[0136] Based on these exemplary data, the composition of a cell culture medium with reduced deviation from the reference total Mn impurity level is determined by an optimization algorithm. For example, it can be determined that a cell culture medium consisting of 80% of A2 and 20% of B1 has a reduced deviation from the reference total Mn impurity level, which is 1.25 ppm. In contrast, a cell culture medium consisting of 80% of A1 and 20% of B2, for example, has a total Mn impurity level of 3.60 ppm, which is a greater deviation from the reference total Mn impurity level of 1.25 ppm and exceeds the upper limit of 1.32 ppm. Therefore, batches A2 and B1 can be determined as preferred batches of raw materials A and B in the cell culture medium, respectively, for example.

[0137] In another example, the method according to the present disclosure can be used to optimize the composition of raw materials in a cell culture medium comprising a plurality (three) of raw materials A, B and C. For raw materials A and B, two batches are available for each (referred to as batches A1, A2, B1, B2, respectively, similar to Figure 2a ). For raw material C, three batches are available, referred to as C1, C2, C3, respectively. In addition, for each of raw materials A, B and C, a reference batch associated with a commercially available cell culture medium is available, referred to as “Aref”, “Bref” and “Cref”, respectively.

[0138] For each batch A1, A2, B1, B2, C1, C2, C3, Aref, Bref and Cref, the impurity level of the functional impurity manganese (Mn) and the impurity level of another impurity copper (Cu) are obtained (e.g., by determining and / or retrieving the impurity levels from a database), as shown in Figure 2b For the cell culture medium, a recipe is obtained that shows that the cell culture medium consists of 40% of raw material A, 30% of raw material B and 30% of raw material C. As an example, it can be assumed that the weights of these impurities are as follows (e.g., according to their functional importance): Mn 0.7, Cu 0.3. Based on these exemplary data, it is determined that the reference total Mn impurity level of the reference cell culture medium consisting of the raw materials of the reference batches Aref, Bref and Cref is 0.88 ppm, and the reference total Cu impurity level is 0.67 ppm, resulting in a reference total weighted impurity level of 0.817 ppm.

[0139] Based on these exemplary data, a cell culture medium composition is determined (e.g. by an optimization algorithm) for which the deviation from the reference total Mn impurity level and the reference total Cu impurity level is reduced (e.g. minimized). For example, a cell culture medium consisting of 40% of A2, 30% of B1 and 30% of C3 can be determined for which the deviation from the reference total Mn impurity level and the reference total Cu impurity level is reduced, with a total Mn impurity level of 0.975 ppm, a total Cu impurity level of 1.175 ppm and a total impurity level of 1.035 ppm. In contrast, for example, a cell culture medium consisting of 40% of A1, 30% of B2 and 30% of C1 has a total impurity level of 1.99 ppm, which is a larger deviation from the reference total impurity level of 0.817 ppm. Thus, batches A2, B1 and C3 can be determined as preferred batches of raw materials A, B and C, respectively, in a cell culture medium.

[0140] In comparison to the exemplary data sets in Figure 2a and 2b , the number of raw materials, impurities and batches that can be input into a cell culture medium according to the method of the present application is significantly higher. In this case, it can be advantageous to determine the individual contribution of one or more raw materials to a certain total impurity level and then only consider those raw materials that contribute more than a predefined threshold to the total impurity.

[0141] In one non-limiting example, when optimizing a cell culture medium, only those impurities in that contribute at least a certain minimum percentage to the total impurity level in the target culture medium may be considered (referred to as "traits" in the following example) . This can be expressed, for example, by the sensitivity matrix of the medium , where : : , where denotes the mass fraction of a raw material in the medium for a powder formulation, while denotes the mass fraction of a raw material in the medium for a liquid formulation. Here, i denotes the average of the impurity profile k of a raw material , where the average is determined by a representative set of reference batches of the raw material, but is not limited thereto. For example, the predefined threshold may be set to 0.05. In the example of a liquid formulation, the summation range can be extended to a set of materials and replaced by ,in This refers to the corresponding mass fraction of the liquid culture medium. In a further example of optimizing the overall composition of cell culture media, if a certain raw material meets the aforementioned criteria for at least one of the culture media in the composition, then its use can be considered, for example. This approach can, for example, ensure that as few raw materials as possible are subject to specification restrictions that would otherwise create additional work during the production and procurement process.

[0142] Regarding the deviation between the reference total impurity level and the determined total impurity level, such as the reference... Figure 2a and Figure 2b As described above, a non-limiting example for comparing these total impurity levels (referred to as "trait" in the following examples) will be provided below.

[0143] The following formula can be used to express the deviation between a given total impurity level (e.g., referred to as the simulated trait) and a reference total impurity level (e.g., referred to as the reference trait): .

[0144] For example, when determining the relative error, divide by This can be optional. In another example, for a given cell culture medium (e.g., medium A), the objective function is... The value can be optimized (e.g., maximized or minimized). For example, for the value to be analyzed... For each type of impurity, calculate the impurity level in the reference culture medium. (For example, it could be the impurity level of the impurity in the reference culture medium, or the upper and / or lower limits of the impurity level) and the impurity level of the impurity in the simulated or calculated cell culture medium. The deviation between the two is squared to generate the sum of absolute errors of the impurities. For example, this can be achieved by multiplying the sum of absolute errors by a weighting factor. We then weight each impurity. The (optionally weighted) absolute errors of all the impurities to be analyzed are then summed to generate the objective function. The value of .

[0145] For example, based on the given exemplary objective function above, the method of this disclosure aims to when Impurities corresponding to the reference culture medium When the impurity level is [high], Minimize the value; or when Impurities corresponding to the reference culture medium When the predetermined upper and / or lower limits of the impurity level are met, Maximize the value of (e.g., maximize the difference between the upper and / or lower bounds).

[0146] For example, in calculating the given exemplary objective function above... When calculating the value, the following formula can be used: .

[0147] According to this exemplary formula, specific impurities in the simulated cell culture medium Total impurity level equal to the culture medium It contains multiple ( The specific impurity in the raw material The sum of impurity levels. More specifically, it is necessary to consider not only multiple ( The levels of each impurity in the raw materials (of each type) should be considered, as well as the levels of each impurity in different batches of each raw material. The decision on whether to include a specific batch of a particular raw material in the cell culture medium to be optimized should be based on... It can be 0 (i.e., excluding the batch) or 1 (i.e., including the batch). For example, when minimizing the objective function, the preferred batches in each raw material with each impurity level can be determined, as will be discussed in detail below. Figure 8 and Figure 9 Further description.

[0148] For example, based on the exemplary objective function given above, the method of this disclosure can be applied not only to optimize the selection of raw materials for one or more impurities in a single cell culture medium, but also further to optimize multiple cell culture media (e.g., culture medium combinations) having various culture medium formulations. For example, the following formula can be used to optimize the selection of raw materials for one or more impurities in a single cell culture medium. Joint optimization of the objective function of cell culture medium: , Among them, factors It can represent the optional weight of a specific culture medium in the combination.

[0149] Figure 3 A flowchart 300 illustrating an exemplary embodiment of the method according to this disclosure is shown. Without limiting the scope of this disclosure, it can be assumed that actions 310 to 340 in flowchart 300 can be derived from reference to... Figure 21 The device shown is used to perform this action.

[0150] Action 310 is to obtain the level of each impurity of at least one impurity in each of a variety of raw materials.

[0151] Consider for reference Figure 2a and 2bFor the described example plurality of raw materials, we obtain a respective impurity level of impurity Mn in each raw material A, raw material B, and raw material C. In addition, we obtain a respective impurity level of at least one other impurity Cu in each raw material A, raw material B, and raw material C. Methods of obtaining these impurity levels can include, for example, retrieving the impurity levels from a raw material database, or determining at least one of these impurity levels (e.g., by instruction and / or control). For example, consider the following Figure 21 The illustrated device performs act 310, and the device communication interface can be used to obtain the respective impurity levels (e.g., by obtaining the respective impurity levels from another device, such as a database device or a determination device).

[0152] Act 320 is obtaining a recipe for a cell culture medium, where the recipe shows proportions of a plurality of raw materials included in the cell culture medium.

[0153] Consider the example described with reference to Figure 2a and 2b For the described example, a recipe for a cell culture medium is obtained, where the recipe shows proportions of a plurality (two) of raw materials A and B included in the cell culture medium as 80%:20% (see Figure 2a ). In another example, the recipe shows proportions of a plurality (three) of raw materials A, B, and C included in the cell culture medium as 40%:30%:30%. Obtaining a recipe for a cell culture medium can include, for example, retrieving the recipe from a database of culture medium recipes.

[0154] Act 330 is determining a total impurity level of at least one impurity in the cell culture medium based at least in part on the recipe and the respective impurity levels.

[0155] Consider the example described with reference to Figure 2a and 2b Based on the respective impurity levels of each raw material, a total impurity level of impurity Mn and / or at least one other total impurity level of impurity Cu in a cell culture medium produced according to the respective recipe can be determined. Here, the respective impurity levels in each raw material can refer to respective batches of the raw materials (e.g., batches Al, A2 of raw material A, or batches Bl, B2 of raw material B).

[0156] In another non-limiting example of act 330, a total impurity level of impurity Mn and / or at least one other total impurity level of impurity Cu in a cell culture medium produced according to the respective recipe can be determined according to the following formula: Cell culture medium including a plurality of raw materials in powder form impurities : .

[0157] In another non-limiting example of act 330, a total impurity level of impurity Mn and / or at least one other total impurity level of impurity Cu in a cell culture medium produced according to the respective recipe can be determined according to the following formula: (​ Cell culture medium in liquid form of raw materials Impurities in : .

[0158] In these examples, is a raw material Impurities in are mass fractions (e.g. impurity levels) of the raw material are mass fractions (e.g. as indicated by the formulation of the cell culture medium ) of the raw material in the cell culture medium , and are concentrations (e.g. in g / L, as further indicated by the formulation of the cell culture medium ) of the cell culture medium powder in the liquid cell culture medium.

[0159] For example, information indicative of the respective total impurity levels of Mn and / or Cu can be output as a result of action 330. Considering that action 330 is performed by an apparatus as indicated in Figure 21 , an apparatus user interface can be used to output the respective total impurity levels (e.g. by presenting the total impurity levels in the form of a visual user output on a screen).

[0160] In one example, the respective contributions of the raw materials mentioned in Figure 2a and 2b to the total impurity levels of Mn and / or Cu can be determined, e.g. this can comprise determining a sensitivity matrix as further described with reference to Figure 2a and 2b .

[0161] Optional action 340 determines whether the total impurity level and / or at least one other total impurity level meets at least one predefined criterion. For example, action 340 can comprise determining whether the total impurity level and / or at least one other total impurity level in the cell culture medium is below a predefined upper limit, and / or determining whether the total impurity level and / or at least one other total impurity level in the cell culture medium is above a predefined lower limit.

[0162] Considering the examples described with reference to Figure 2a and 2b , it can be determined whether the total impurity levels of Mn and / or Cu in the respective cell culture media are below a predefined upper limit and / or above a predefined lower limit of the respective impurities in the respective cell culture media (e.g. by comparing the determined total impurity levels with the respective upper limit and / or lower limit).

[0163] In one non-limiting example, it can be assumed that at least one total impurity level of a toxic impurity and at least one total impurity level of a functional impurity are determined in act 330, and that it is determined in act 340 whether these two impurities meet the predefined criteria. In such an example, act 340 can comprise determining whether the total impurity level of the toxic impurity is below a predefined upper limit, and whether the total impurity level of the functional impurity is above a predefined lower limit and below a predefined upper limit.

[0164] In another non-limiting example of optional act 340, it is determined, based at least in part on the upstream process model and / or the downstream process model of the cell culture medium, whether the total impurity level and / or the at least one other total impurity level meets the at least one predefined criterion. For example, each total impurity level determined in the cell culture medium can be multiplied by additional factors representing the downstream process before determining whether each total impurity level meets the at least one predefined criterion (e.g., upper limit and / or lower limit). For example, these additional factors according to the downstream process model can be considered together with the weighted sum of different media according to the upstream process model.

[0165] Further considering, for example, the upstream process model, it can be assumed a scenario for a total impurity level of a specific impurity in the cell culture medium, in which during the upstream process any compound uptake of the impurity by the cells in the bioprocess is explicitly neglected (e.g., this can be achieved based on standard mass balance approaches). Furthermore, any possible dilution of the impurity during the upstream process can be neglected (e.g., due to the addition of acids, bases or antifoam). For example, a mixture of media can be considered (considering, for example, a fed-batch process with a base medium and a feed medium, e.g., considering a weighted combination of these media), such that a total impurity level after the upstream process comprising sub-processes can be determined .

[0166] Considering, for example, the downstream process model, an expected dilution of a specific impurity in the cell culture medium can be considered. Since most downstream processing steps can lead, for example, to a dilution of the impurity, the level of this impurity in the finished drug product can typically be lower than the total impurity level in the cell culture medium. Thus, for the total impurity level in the cell culture medium, a more relaxed upper limit and / or lower limit can be set. For example, after considering the upstream and downstream model processes, the impurity level (“trait”) can be expressed as: , wherein the downstream process comprising sub-processes is considered.

[0167] Figure 4 A flowchart 400 illustrating another exemplary embodiment according to the methods of the present disclosure is shown. For example, as referenced above, the flowchart 400 can be used to determine whether a total impurity level of a specific impurity in a cell culture medium meets at least one predefined criterion. Figure 3The action 410 can be performed after the action 330 or after the action 340. Without limiting the scope of the present disclosure, it can be assumed that the action 410 in the flowchart 400 can be performed by the apparatus 100 shown in Fig. 1. Figure 21

[0168] The action 410 determines an allowed range for the impurity level of at least one impurity in at least one raw material of the plurality of raw materials based at least in part on the recipe (e.g., the recipe obtained in the action 320), the respective impurity level of the at least one impurity in each raw material of the plurality of raw materials (e.g., the impurity levels obtained in the action 310), and the at least one predefined criterion (see the action 340). Herein, the allowed range for the impurity level of an impurity in a raw material can for example be understood as a numerical range for the impurity level of the impurity in the raw material, which impurity level can vary within the numerical range while the total impurity level of the impurity in the cell culture medium still satisfies the predefined criterion (e.g., the upper limit and / or the lower limit). In other words, the total impurity level of the at least one impurity in the cell culture medium determined based at least in part on the recipe and the (e.g., any) impurity level of the at least one impurity in the at least one raw material within the determined allowed range satisfies the at least one predefined criterion (e.g., satisfies the at least one predefined criterion with a certain probability).

[0169] In one non-limiting example of the action 410, determining the allowed range for the impurity level of at least one impurity in at least one raw material of the plurality of raw materials can not be based on the respective impurity level of the at least one impurity in each raw material of the plurality of raw materials, but on the respective impurity level in one or more raw materials which contribute more than a predefined threshold to the total impurity level. This can for example reduce the optimization effort when determining the respective allowed range.

[0170] Consider the example described with reference to Figure 2a Based on the recipe of the respective cell culture medium comprising the raw materials A and B, the respective impurity level of the impurity Mn in each raw material A and B (e.g., for each batch of raw material A and B, respectively), and the predefined criterion that the total impurity level of Mn in the cell culture medium should be above the lower limit and / or below the upper limit, it can be determined a respective allowed range (e.g., with reference to a reference cell culture medium in which the total impurity level of Mn is allowed to fluctuate + / - 20% around the reference, which reference cell culture medium can for example consist of batches Aref and Bref of raw materials A and B, respectively) within which the impurity level of Mn in the raw materials can vary while still satisfying the predefined criterion for the overall cell culture medium. Furthermore, it can be determined an additional allowed range for the impurity level of Mn in raw material B, and / or it can be determined an additional allowed range for other total impurity levels (e.g., Figure 2b the impurity levels of Mn and Cu in the raw materials A, B, and C in the cell culture medium shown in Fig. 2). ​

[0171] In another non-limiting example of act 410, determining the respective allowable range of the impurity level of the at least one impurity of the at least one raw material of the plurality of raw materials can include calculating a probability that the total impurity level of the at least one impurity in the cell culture medium does not satisfy at least one predefined criterion based at least in part on an initial range of the impurity level of the at least one impurity of the at least one raw material, and determining the respective allowable range of the impurity level based at least in part on the calculated probability and the initial range of the impurity level, particularly by using an optimization algorithm.

[0172] With reference to the examples described in Figure 2a , it can be assumed that an initial range of the impurity level of Mn in the raw material A (e.g., ±20% of the impurity level Aref of Mn in the raw material A), where the initial range can be based, for example, on statistical variations of the impurity level in the raw material A (e.g., by considering multiple batches of the raw material A). Subsequently, a probability that the total impurity level of Mn in the cell culture medium can not satisfy at least one predefined criterion (e.g., a deviation of the determined total impurity level of Mn from a reference total impurity level of Mn in the cell culture medium is greater than an upper limit) can be determined (e.g., a probability of out-of-specification of the target medium). This probability can be compared to a threshold probability, and if the probability is greater than the threshold probability, the initial range can be narrowed to determine the allowable range. In another example, if the probability is less than the threshold probability, the initial range can be expanded to determine the allowable range (e.g., a probability of out-of-specification of the target medium). At this point, the probability of a violation that the impurity level of the cell culture medium remains within an acceptable bi- faceted property band can be represented). This adjustment of the initial range can be repeated multiple times as part of the optimization algorithm to determine the allowable range (see below ). Figure 6

[0173] Figure 5 A flowchart 500 illustrating another exemplary embodiment of a method according to the present disclosure is shown. For example, the acts of the flowchart 500 can be performed after act 330 or after act 340 described with reference to Figure 3 , or as part of act 410 described with reference to Figure 4 . Without limiting the scope of the present disclosure, it can be assumed that the acts of the flowchart 500 can be performed by the apparatus described with reference to Figure 21 .

[0174] ​​In the first action 510, the mean and standard deviation of specific impurity levels for each impurity in the raw materials (“raw material characteristics”) can be calculated based on the raw material database 570. These calculated values ​​can be used to determine an initial range of impurity levels for each impurity in each raw material. Subsequently, those raw materials among the various raw materials contained in the cell culture medium that make a specific contribution to the total impurity level of interest in the cell culture medium (“sensitive raw materials”) can be identified (see action 520, for example, by referring to...). Figure 2a and 2b The sensitivity matrix mentioned above. Identification of sensitive raw materials can be based on the corresponding formulation of the cell culture medium (“culture medium ingredient list”), which can be obtained from the culture medium formulation database 580, along with predefined criteria (e.g., upper and lower limits) regarding the total impurity levels in each cell culture medium (see action 530, “Matching Feature Target Specification Range”). Subsequently, when optimizing the initial range of impurity levels obtained from the raw material database, only the impurity levels in these sensitive raw materials can be considered, while the calculation of the total impurity feature content of the culture medium considers the impurity content of all raw materials in the formulation (see action 540). This may involve optimization algorithms to ensure probabilistic... The probability of being equal to or below a predefined threshold. Based on these optimized raw material limits for specific impurity levels (see Action 550), the raw material database can be updated to ensure more robust raw material procurement (see Action 560).

[0175] Actions 510 to 560 shown in flowchart 500 can be understood, for example, as an optimization algorithm capable of tracing back to the raw material level using input data for the culture medium level. Generally, this optimization algorithm can utilize the expected acceptable levels of impurities in the cell culture medium and data from different batches of raw materials in the medium to find optimal values ​​for each raw material and each impurity to minimize violations of specific, predefined standards or limits in the cell culture medium for their respective impurity levels. For example, inputting culture medium specifications, culture medium ingredient lists, and limited assay data for raw material batches, and as output, the optimization algorithm provides specifications (i.e., allowable ranges) for the raw material levels. Specifically, an exemplary overall objective could be to optimize raw material specification limits to ensure that the cell culture medium meets predetermined specification requirements (e.g., the probability of compliance is equal to or higher than a desired threshold probability).

[0176] For example, actions 510 to 560 shown in flowchart 500 can be understood, for instance, as part of a self-learning and continuously improving system. The procedure can be repeated periodically, for example, when new materials, new batches of raw materials, or new culture medium formulations are added to the steps, to obtain a finer and more robust range.

[0177] Figure 6A flowchart 600 illustrating another exemplary embodiment of the method according to this disclosure is shown. For example, the actions of flowchart 600 may be performed as part of determining the allowable range of various impurity levels in the raw materials, such as... Figure 4 As described in action 410 of the flowchart 400 shown, and as part of optimizing raw material limits, as... Figure 5 As shown in flowchart 500. Specifically, the actions of flowchart 600 can be understood as a sub-part of the method shown in flowchart 500, wherein the actions according to flowchart 600 can be performed after selecting limits only for sensitive raw materials in action 540 according to flowchart 500 and before optimizing raw material limits in action 550. Without limiting the scope of this disclosure, it can be assumed that the actions of flowchart 600 can be derived from references Figure 21 The device shown is used to perform this action.

[0178] For example, considering the initial range of impurity levels in the raw materials (“RM characteristic values ​​from a specified range”), the specification values ​​of the resulting culture medium can be calculated using Gaussian sampling or other sampling methods (see Actions 610 and 620). For example, some values ​​based on a Gaussian distribution can be selected within the initial range (e.g., for reliable statistics). Based on these values, the probability that the total impurity level in the cell culture medium may not meet at least one predefined criterion can be determined. (For example, the probability of out-of-specification (OOS) in the target culture medium) (see Action 630). For example, if the determined total impurity level in the cell culture medium deviates from the reference total impurity level (e.g., represented by the objective function) If the result exceeds the upper limit, at least one predefined criterion may not be met. The following pseudocode provides an example of loop counting for oversized results:

[0179] In this example, the probability of violation can be based on Calculation. Then, it can be checked whether the probability of violation is lower than a predefined probability threshold (i.e., ).For example, This means that at least 95% of the produced culture media batches are expected to meet the required specifications. Subsequently, when the probability of non-compliance is below a predefined probability threshold, the permissible range of each impurity level in the raw materials can be determined, for example, by calculating and minimizing an objective function, which includes weighted factors. (For example, The weighted probability of violation, and a penalty term used to ensure that the optimization algorithm calculates the widest possible range of impurity levels for the optimized raw materials (e.g., instead of deriving a minimum allowable range to minimize the deviation of the total impurity level from the reference total impurity level, see action 640). Then, the reference...Figure 5 The above raw material database can be updated with the respective raw material impurity level limit within the allowed range. For example, Figure 6 The illustrated actions can be referred to as a CMA-ES loop, which can be repeated for a number of generations (G) and further restarted a number of times (R) with different initial ranges to address potential local minimum problems.

[0180] The exemplary loop count of the hyper-specification result given by the above pseudo code can be understood as an inner loop, which can be part of an optimization algorithm that can calculate the mean and standard deviation of the respective characteristics of each raw material using the batch raw material characteristic data of the impurity levels, for example, by calculating the mean of the different batches and the corresponding medium characteristics values. Then, the inner loop can be executed to calculate (e.g., to find) the percentage of hyper-specification media, which can correspond to the above described by defining a function as a measure that counts hyper-specification events.

[0181] Further regarding the inner loop workflow, the objective function can also be designed such that a given medium characteristic (i.e., a specific total impurity) can be tracked if it is below or above a defined medium characteristic specification range (e.g., according to predefined standards for cell media). In this exemplary evolutionary strategy setup, the optimization algorithm can try to minimize the objective function value (fitness value), which directly depends on the probability of a violation, by optimizing / adjusting the mean and standard deviation of the specification range of a set of raw material given characteristics (i.e., the respective impurity levels in a plurality of raw materials). As a non-limiting example, the respective objective function can be: = min!

[0182]

[0183] In particular, the exemplary objective function depends on which the optimization algorithm tries to minimize. The objective function can sum up the scaled contributions of all medium characteristics to be optimized (i.e., all impurities) like the biological load and the manganese content and sum up for each characteristic in all raw materials that contribute to the total impurity level in the cell medium. The penalty term containing the sigma expression can be beneficial to keep the raw material specification ranges as broad as possible, which can be desirable from a business perspective, for example, while ensuring that the upper limit given by the acceptable probability of a violation (i.e., the maximum percentage of batches in the production of the medium that are expected to be hyper-specification) is adhered to.

[0184] Violations of the media specifications can occur in different scenarios, either unidirectional (above upper limit / below lower limit) or bidirectional. The optimization algorithm can provide different handling based on the different scenarios. In the former case, one can optimize (i.e., adjust ) for the target; in the latter case, one can only optimize for the target. This switching strategy can help the convergence of the algorithm if one of the scenarios can sufficiently reduce the violation probability. Once the optimization is successful, the optimized specification range for each raw material is calculated as follows.

[0185]

[0186] In particular, without loss of generality, one can assume equals 3. For example, for a normally distributed random feature variation in a raw material, a range of 3 will result in 99% of the purchased raw material batches meeting this specification requirement.

[0187] Figure 7a , 7b and 7c show another exemplary embodiment according to the method of the present disclosure. For example, Figures 7a to 7c may involve an exemplary demonstration of the concept according to the method of the present disclosure, as further described with reference to Figure 4 , 5 and 6.

[0188] A commonly used cell culture medium formulation, i.e., RPMI open source medium with slight modifications as shown in Figure 7a , can be used as input to illustrate the application of the algorithm as described with reference to Figure 4 , 5 and 6. The exemplary medium contains 38 compounds (i.e., raw materials) with two selected feature values (i.e., impurity levels) for each of two features (i.e., impurities), manganese and nickel. Figure 7b shows the convergence status (i.e., fitness value) at different number of restart iterations for the evolutionary strategy in the optimization process. Here, each restart can be considered as an independent optimization simulation. As shown in Figure 7b , in the first 15 restarts, the algorithm starts from optimizing and adjusting . In this case, for some settings, the violation probability can be significantly reduced, but still not enough to meet the acceptance criteria (e.g., violation probability below 10%) because some raw materials still result in exceeding and not meeting the target specification range. Therefore, one can switch the strategy to further optimize from only the standard deviation of the raw material features. The best convergence occurs at the 18th restart, when has been optimized. In Figure 7cA summary of the algorithm application for some sensitive compounds according to this example is provided. In this example, the algorithm attempts to reach a global minimum by adjusting and for different raw materials, most importantly, as shown in Figure 7c , for both sensitive compounds, the and are reduced in both cases, thus reducing the probability of a violation.

[0189] Figure 8 A flowchart 800 illustrating another exemplary embodiment of the method according to the present disclosure is shown. For example, the action 810 can be performed after the action 330 or after the action 340 as described with reference to Figure 3 . Without limiting the scope of the present disclosure, it can be assumed that the action 810 in the flowchart 800 can be performed by the apparatus as described with reference to Figure 21 .

[0190] The action 810 determines a preferred batch of at least one raw material of the plurality of raw materials, wherein the deviation of the total impurity level and / or at least one other total impurity level in the cell culture medium from the respective reference total impurity level in the cell culture medium is reduced, at least partially based on the impurity level of at least one impurity of the at least one raw material of the preferred batch, in particular by minimizing the deviation of the total impurity level and / or at least one other total impurity level from the respective reference total impurity level using an optimization algorithm.

[0191] For example, minimizing the deviation from the respective reference total impurity level can be understood as meaning that the deviation is controlled to be within a range around the total reference impurity level.

[0192] Considering the example as described with reference to Figure 2a , the batch A2 can be determined as the preferred batch of the raw material A, wherein the deviation of the total impurity level of Mn in the cell culture medium from the reference total impurity level of Mn is reduced (e.g., reduced compared to when the total impurity level of Mn in the cell culture medium is determined using the batch Al). For example, determining the preferred batch of the raw material can be understood as meaning that the preferred batch is determined based on minimizing (e.g., at least locally minimizing) the deviation of the total impurity level from the respective reference total impurity level. For a sufficient number of the plurality of raw materials, determining the preferred batch of the raw material can require using an optimization algorithm, which will be further described below.

[0193] As described with reference to Figure 2a and 2b , the following objective function can be used to represent the deviation between the determined total impurity level (e.g., referred to as a simulated feature) and the reference total impurity level (e.g., referred to as a reference feature).

[0194]

[0195] wherein, the value of

[0196] For a batch of a particular raw material to be optimized for inclusion in a cell culture medium, the value of may be 0 (i.e., the batch is not included) or 1 (i.e., the batch is included). For example, for two raw materials, each with three batches, the set of cell culture media [A1 A2 A3 B1 B2 B3] can be used to represent the batches of raw materials included in the cell culture medium, where the set [1 0 0 0 1 0] can represent the inclusion of batch A1 of raw material A and batch B2 of raw material B in the cell culture medium. In one example, the preferred batch of raw material A can be determined by calculating the value of for all possible batch combinations [A1 A2 A3 B1 B2 B3] of the cell culture medium (e.g., for two raw materials, each with three batches, there are nine combinations). The preferred batch of a particular raw material can then be determined from the batch combination that minimizes the value of . To improve the efficiency of this approach in evaluating a large number of raw materials (e.g., 50 raw materials) and their respective batches in a cell culture medium, rather than considering each of the plurality of raw materials (see for the number of raw materials in a cell culture medium), only those raw materials whose contribution to the respective total impurity level is greater than a predefined threshold are considered.

[0197] Another approach to improve efficiency in determining the preferred batch can use a global optimizer instead of determining for each possible batch combination. For example, this global optimizer can employ a customized Covariance Matrix Adaptation Evolution Strategy (CMA-ES, e.g., developed from an evolutionary strategy known in the literature), the specifics of which will be further described with reference to Figure 8 .

[0198] Figure 9 A scheme illustrating another exemplary embodiment of a method according to the present disclosure is shown. For example, the method according to Figure 9 may be part of determining the preferred batch, as described in act 810 of flowchart 800. Figure 8

[0199] As described with reference to Figure 8 , a customized CMA-ES optimization algorithm can be employed to minimize ​​Since evolutionary optimization algorithms like CMA-ES are typically used to solve continuous optimization problems, the transformation function 920 can be used to solve discrete problems (e.g., selection of preferred batches) as part of a customized CMA-ES 900.

[0200] Reference can be made to Figure 9 understand an exemplary application of the transformation function 920. At each iteration of the CMA-ES optimization algorithm 900, the provisional result of the optimization algorithm can be given by the variable in the form of a tensor 910. Then, the transformation function 920 can map to the corresponding values , where denotes the selection of a particular batch. In particular, is a continuous variable representing the selection of a batch in the interval [1, n batches ]. The transformation function 920 as a mapping function then ensures that continuous values in the interval are mapped to the respective integer batch numbers 930 with equal probability. The term before” denotes the variable generated by the previous iteration of the algorithm.

[0201] Figure 10a and 10b show exemplary experimental data according to the present disclosure.

[0202] As a non-limiting example of a cell culture medium to be optimized, we examined a fed-batch medium for the cultivation of Chinese hamster ovary (CHO) cells. CHO cells were provided that were genetically engineered to express a certain product (in this case an antibody). A batch of cell culture medium was inoculated with CHO cells at a number of 3 x 10 5 cells per milliliter of cell culture medium and subjected to a fed-batch cultivation for a period of 14 days in a cell culture incubator under conventional cell cultivation conditions (e.g., 36.8°C ± 0.2°C, 7.5% ± 0.5% CO2, 110 rpm ± 5 rpm). From day 3 of cultivation, the viable cell concentration (VCC) was determined daily using a cell counter (e.g., Vi-CELL XR Cell Viability Analyzer Beckman Coulter). In addition, from day 7 of cultivation, the product titer (i.e., the amount of antibody produced per volume of cell culture medium) was determined by double-layer interferometry (e.g., using an Octet device Sartorius Molecular Devices).

[0203] Figure 10a show exemplary experimental data according to the present disclosure. Figure 10aperformance comparison of the reference medium and the medium composed of the supplier A. Herein, Figure 10a The left side shows that both the reference medium and the medium of supplier A provided a viable cell concentration (VCC) of up to 200 x 10 5 cells / mL cell culture medium at day 7 of cultivation. However, the cell concentration decreased over time as the cell culture period was extended beyond day 7. Figure 10a The right side shows that the product titer in the cell culture medium of supplier A was about 20% lower than the product titer in the reference medium after 14 days of cultivation.

[0204] To improve the cell culture medium of supplier A, we determined the impurity level of at least one impurity per raw material in the cell culture medium of supplier A. In addition, we also obtained the formulation of the cell culture medium of supplier A. Based on these data, we determined the total impurity level of at least one impurity in the cell culture medium of supplier A and further determined whether this total impurity level met the predefined criteria. The results showed that the concentration of the functional impurity "Element A" in the cell culture medium of supplier A was lower than the total impurity level of Element A in the reference medium, wherein the total impurity level of Element A in the reference medium was in the range of 0.1 to 0.2 mg / L. Figure 10b This is shown on the left side with a horizontal dashed line.

[0205] In addition, by performing the method according to the present disclosure, it was determined that the raw material A ("RM A") contributed the most to the deviation between the total impurity level of Element A in the cell culture medium of supplier A and the total impurity level of Element A in the reference medium. Subsequently, a preferred batch of RM A was determined using the method according to the present disclosure. The impurity profile of this preferred batch resulted in a total impurity level of Element A that was higher than the total impurity level of Element A in the reference medium. In this example, the reduction of the deviation was achieved by exceeding the total impurity level of Element A in the reference medium. This is shown on the left side with a horizontal dashed line. Figure 10b The concentration of Element A in the cell culture medium of supplier A is shown to be lower than the total impurity level of Element A in the reference medium, whereas the concentration of Element A in the optimized cell culture medium of supplier A comprising the preferred batch of RM A (referred to as "Supplier A + new source RM A" in the right side Figure 10b is higher than the total impurity level of Element A in the reference medium (i.e., thus meeting the predefined criteria).

[0206] In a subsequent experiment performed under similar conditions as described above for the reference Figure 10a medium, the product titer was determined every 24 hours starting at day 7. The right side Figure 10b shows the resulting product titers. Here, the results show that the cell culture medium of supplier A (referred to as "Supplier A" on the right side Figure 10btermed "Supplier A media (old source RMA)" yielded the lowest titers after 13 days of cultivation, while the cell culture medium optimized according to the method of the present disclosure (on the right side Figure 10b termed "Supplier A + new source RMA" yielded higher product titers than observed with the reference medium.

[0207] These experimental data show that the method according to the present disclosure is capable of improving the performance of a cell culture medium, e.g. the productivity of cells cultivated in an optimized cell culture medium is increased as reflected by an increased product titer compared to a cell culture medium not applying the method.

[0208] Figure 11 Further exemplary experimental data according to the present disclosure are shown.

[0209] As a non-limiting example, the method of the present disclosure will be described Figure 10a and 10b The method of the present disclosure described is compared to a method known in the art for improving a cell culture medium. It is a known method to supplement a cell culture medium with trace elements to improve cell culture performance. To this end, in the comparative use experiment, a mixture of elements comprising element A and other elements is supplemented to the cell culture medium of Supplier A. These selected elements are suggested based on the contribution of the impurity levels of RMA to these elements according to the method of the present disclosure (see Figure 11 : "Supplier A media (mixture of elements)").

[0210] Figure 11 Product titers after 13 days of cultivation are shown. In particular, the product titer obtained with the cell culture medium containing the new source RMA is higher than with the reference medium (see Figure 11 : "Supplier A media (new source RMA)"). At the same time, the medium obtained by supplementing the cell culture medium of Supplier A with element A and other elements ("Supplier A media (mixture of elements)") exhibits product titers after 13 days of cultivation comparable to the reference medium. These findings show that either replacing RMA or supplementing the mixture of elements suggested according to the method of the present disclosure can improve product titers.

[0211] Figure 11 The examples shown demonstrate that the quality of cell cultivation can be optimized by using the method according to the present disclosure as evidenced by the high cell culture product titers obtained with the cell culture medium optimized using the method of the present disclosure.

[0212] Figures 12 to Figure 15 Further exemplary experimental data according to the present disclosure are shown. In particular, Figures 12 to Figure 15 Exemplary experimental data on an upstream process model are shown.

[0213] Integrated calculation of net elemental concentrations of media formulations using control volume simulation

[0214] According to this disclosure, the control volume simulation presented herein as part of an impurity calculation platform can be considered an important component of upstream process simulation. This simulation can include all supplementary formulations (e.g., culture medium, feed, defoamer, acid / base for pH control) entering a typical culture process (e.g., fed-batch, perfusion, or other types) and substances removed from the process (e.g., process sampling, harvest stream). Since the consumption rate of elements by CHO cells is not considered, the simplified model is independent of any cell-specific element uptake and still provides reasonable predictions because it constitutes an upper limit for impurity accumulation in the upstream process, i.e., a worst-case assessment.

[0215] Figures 12a to 12d Further exemplary experimental data according to this disclosure are shown. Reference Figures 12a to 12d The data shown compares the measured and simulated concentrations of magnesium (Mg), calcium (Ca), cobalt (Co), and nickel (Ni) at a specific time point in the process (e.g., day 12). The simulated values ​​represent the concentrations of trace elements in the basal medium and feed. These results demonstrate that the algorithm can predict the dynamic levels of specific elements (e.g., magnesium, calcium, cobalt, nickel) during upstream processes, and most importantly, predict their final levels at the end of the corresponding upstream process (or culture).

[0216] Simulation of specific elements compared to experimental determinations

[0217] Figures 13a to 13d Further exemplary experimental data according to this disclosure are shown. Specifically, simulated trace metal levels of calcium (Ca), magnesium (Mg), cobalt (Co), and nickel (Ni) in the upstream process were compared with measurements in cell-free culture supernatant from actual experiments (see [link to relevant documentation]). Figures 13a to 13d These exemplary data represent the simulation's ability to predict the elemental composition levels or their upper limit approximations during the process.

[0218] like Figures 13a to 13d As shown, the predictive ability of simulations for daily levels of trace elements in upstream processes can vary by element because the degree to which elements are absorbed by cells during the process differs (not measured here). Cobalt is the most obvious example in this case, with its daily measured levels matching the simulated levels (virtually no absorption by cells). For other elements such as calcium and magnesium, on certain days in the process, when they are heavily absorbed by cells, the results deviate from the simulated total content change trajectory. Despite these differences, on day 12 (near the end of the process), the simulated levels in both cases again closely approximate the measured supernatant values ​​(see [link to data]). Figures 12a to 12dand 13a to 13d). This highlights the effectiveness of the model approach as an upper limit estimate. The advantage of this approach is its robustness, which allows meaningful predictions of maximum impurity levels in a liquid without explicit knowledge of the uptake rates of a particular cell line or organism.

[0219] Closed loop optimization enables adjustment of raw material specifications to meet limit requirements in USP and DSP

[0220] It is further elucidated that the platform according to the present disclosure is able to calculate the expected total impurity content and the contribution of different media used in the upstream process and compare it to a target value (e.g. maximum upper limit, trace metal content at a given time in the process should remain below the maximum upper limit). This information can be used to further optimize the raw material selection step, as it is required to ensure this limit value by the combined use of media, rather than performing an "each media" optimization. Thus, the joint optimization using the upstream process model can increase additional degrees of freedom (by using an optimization algorithm) in two ways: (a) possibly relaxing or (b) tightening the allowed impurity specification range of certain raw materials. In this way, the method simplifies a robust procurement process and enables the determination of suitable raw material batches for media production for a specific bioproduction process.

[0221] It is further elucidated the integrated relationship between the upstream process model performing forward simulation and the optimization algorithm closing the backward loop, and studied for the example of nickel. The predictive power of the upstream process model is demonstrated by the prediction plot for nickel, where the simulated values establish an upper limit, as shown in Figures 13a to 13d Data representative of day 12, which is close to the end of the process, are incorporated into the optimization algorithm as key parameters. This integration facilitates the refinement of raw material specifications in the combined media use.

[0222] Figure 14 Further exemplary experimental data according to the present disclosure are shown. In particular, Figure 14 The convergence status of the objective function values (i.e. fitness values) of the optimization process in the evolution is shown, thereby depicting the evolution of the fitness values over multiple algorithm restarts, which is a strategy to reduce the risk of getting stuck in local minima. These fitness values serve as a measure to assess the appropriateness and effectiveness of the solutions generated by the optimization algorithm. It is noted that the fitness values close to zero indicate that the algorithm has converged and the deviation remains below a pre-specified threshold (e.g. 5%), thereby highlighting the robustness of the optimization process. The reason for choosing nickel as the focus trace metal is that it was observed that in the selected example formulation, a large fraction of its total amount originates from impurities rather than formal formulation ingredients. This property makes nickel a particularly relevant candidate for optimization here, as it offers a meaningful opportunity to improve the overall process efficiency by reducing the variability caused by impurities.

[0223] Figure 15Further exemplary experimental data according to the present disclosure is shown. In particular, Figure 15 Differential relaxations of various raw material specifications in the hybrid media are shown, and the evolution of raw material specifications after optimization by applying a unified simulation protocol data model is presented. This optimization leads to more relaxed raw material specifications in the hybrid media formulation. However, the degree of relaxation varies from one raw material to another. Raw materials A and B have relatively narrow relaxation bounds as they are classified as sensitive compounds having a high contribution to the media composition. In contrast, raw material C has more freedom in terms of specification relaxation, which reflects its relatively low contribution to the media.

[0224] Figures 16 to 20 Schematic diagrams and further exemplary experimental data according to the present disclosure are shown. In particular, Figures 16 to 20 Schematic diagrams and exemplary experimental data regarding a downstream process model are shown. For example, the downstream process model can predict the characteristic behavior of impurities during the purification process.

[0225] Figure 16 A schematic diagram of an exemplary downstream process according to the present disclosure is shown. The exemplary downstream process can be connected to the upstream process and can for example include a harvest filtration step and a protein A capture chromatography.

[0226] Individual unit operations of the downstream process can be characterized in a simple way by their input / output behavior, which describes the yield and effective dilution of each impurity characteristic by the unit operation. The total characteristic levels reached at the end of the combined upstream process (USP) and downstream process (DSP) can then be summarized as follows:

[0227] As a non-limiting example, the yield (e.g., in amount) of a downstream process step can be written as: , where, V in represents the volume of liquid entering the purification step l , and C in represents the concentration of the characteristic in the liquid. describes the effective yield factor observed for the characteristic l in the process step k . The sum over j describes any additional contribution to the amount of the characteristic k (e.g., an impurity) introduced by process aids (e.g., buffer solutions used for elution or neutralization) in the purification step and still present in the material leaving the purification step. Here, V aid represents the added volume of process aid j in the step, and Process aids j characteristics k concentrations.

[0228] Without limitation, the effective yield of a purification step can be expressed in terms of concentration, i.e.

[0229] where the factor represents the degree of retention of the original volume entering the purification step at the exit of this step.

[0230] For ideal processes, it can be assumed that, e.g., for a filtration step without hold-up or for a capture step like Protein A chromatography where all incoming liquid from the previous step has been removed by washing before elution. In practice, for a real filtration step with hold-up, the value will be smaller than 1 ; while for a real chromatography step, e.g. due to possible liquid hold-up in the dead volume, the value will also be greater than 0.

[0231] In particular, the above approach can be used to describe, e.g., a concentration process step such as a capture chromatography, where the elution volume is typically smaller than the original volume entering the purification step (i.e., ).

[0232] For further simplification, the effect of a washing step can be neglected (in the ideal case, the volume added to the process step equals the volume removed), while its effect on the effective yield of a characteristic k can then be determined empirically by performing suitable experiments beforehand or by consulting the relevant literature. In such cases, the downstream process model can be used to predict the purification cascade outcome for a characteristic k using the above approach, as described below.

[0233] Figure 17 Further exemplary experimental data according to the present disclosure are shown. In particular, Figure 17 spectrum simulations after different process steps are shown, including the upstream process end-point before cell harvest (see "End-point USP" in Figure 17 ), after cell harvest upon filtration completion (see "After filtration" in Figure 17 ), and exemplary downstream process end after the Protein A capture step (see "After Protein A" in Figure 17 ). As an example, magnesium, cobalt, and zinc are used as representative trace metals. The simulation values refer to the simulation results obtained by applying the above downstream process model and using the assay input values (last day of the upstream process).

[0234] Figure 17 This shows the measured trace metal levels at different process steps when upstream process measurements are used as input to the downstream module of the model (see [reference]). Figure 17 The “meas” in the text is different from the simulated values ​​(see [link]). Figure 17 An exemplary comparison between “sim” in the text. From Figure 17 As can be seen, the results generated by the downstream process model are very close to the actual measured values. In this example, the unit operation is considered to be in an ideal state (i.e., assuming that for protein A capture...). It is 0, while for harvest filtering (e.g., 1), and each unit operation The estimated values ​​were derived from trace metal measurements at different stages of the process.

[0235] Figure 18 Further exemplary experimental data according to this disclosure are shown. Specifically, Figure 18 This shows the use of copper in... Figure 17 The corresponding results are obtained using the same simulation settings. From Figure 18 The example shows that after protein A purification, the copper content actually exceeds the level at the upstream process endpoint. Decomposition analysis using the model indicates that this phenomenon is caused by copper impurities in the elution buffer and neutralization buffer used in the protein A capture step (see also...). Figure 19 ). Figure 18 The analysis results show that this trend can also be successfully captured using the downstream modeling method shown.

[0236] Figure 19 Further exemplary experimental data according to this disclosure are shown. Specifically, Figure 19 The decomposition of total copper content after protein A purification, inferred from the measurement results, is shown and compared with model predictions, illustrating the major contribution of the buffer solution used in downstream process steps.

[0237] Figure 20 Further exemplary experimental data according to this disclosure are shown. Specifically, Figure 20 Showing with Figure 17 The same settings were used, but this time the upper limit estimate derived from the upstream process simulation was used as the input to the downstream process model.

[0238] about Figure 19 The data in the middle, we adopted the same... Figure 17 The same method was used, and the same parameter values ​​were used to repeat the experiment, but this time a conservative estimate of the concentration at the end of the upstream process was used as input. Figure 20The results shown indicate that in this case, for characteristics such as magnesium and cobalt, the uptake of biomass is limited relative to the total amount added to the process from the medium, and thus good agreement with the measured values can be achieved. For compounds such as zinc, which are significantly retained in the cell mass, the model naturally produces upper limit estimates, which are particularly evident for the predicted concentrations after the filtration and protein A steps. This shows that the method implemented, starting from the characteristics of the raw materials, provides relevant predictions reproducibly from the raw material characteristics all the way to the results of the upstream and downstream processes. This is a prerequisite for directly linking, for example, the limits for impurity levels in the upstream or downstream processes to the specification range for the raw materials for cell culture medium production.

[0239] Figure 21 A schematic diagram illustrating an exemplary embodiment of an apparatus according to the present disclosure.

[0240] The apparatus 2100 comprises, for example, a processor 2101, and a first memory 2102 as a program and data memory, a second memory 2103 as a main memory, a communication interface 2104, and a user interface 2105 connected to the processor 2101.

[0241] The processor is to be understood as meaning, for example, a microprocessor, a microcontrol unit, a microcontroller, a digital signal processor, an application-specific integrated circuit, or a field programmable gate array. It goes without saying that the apparatus 2100 can also comprise a plurality of processors 2101.

[0242] The processor 2101 executes the program instructions stored in the program memory 2102 and stores, for example, intermediate results and the like in the main memory 2103. The program memory 2102 contains, for example, program instructions of a computer program, for example a computer program according to the present disclosure, which cause the processor 2101 to carry out and / or control the methods disclosed according to the present disclosure when the processor 2101 executes these program instructions stored in the program memory 2102. Furthermore, the program memory 2102 can store, for example, one or more databases, for example a raw material database and / or a medium formulation database, or a representation of one or more databases.

[0243] The program memory 2102 also contains, for example, an operating system of the apparatus 2100, which is at least partially loaded into the main memory 2103 when the apparatus 2100 is started and executed by the processor 2101. In particular, at least a part of the core of the operating system is loaded into the main memory 2103 when the apparatus 2100 is started and executed by the processor 2101.

[0244] Examples of operating systems are Windows, UNIX, Linux, Android, Apple iOS and / or MAC OS operating systems. The operating system, inter alia, allows information and / or data processing using the control device. For example, it manages resources such as main memory and program memory, inter alia, provides basic functions for other computer programs using programming interfaces, and controls the execution of computer programs.

[0245] Program memory (for example) is a non-volatile memory, such as a flash memory, a magnetic memory, an EEPROM (electrically erasable programmable read-only memory) and / or an optical memory. Main memory (for example) is a volatile or non-volatile memory, in particular a random access memory (RAM), such as a static RAM (SRAM), a dynamic RAM (DRAM), a ferroelectric RAM (FeRAM) and / or a magnetic RAM (MRAM).

[0246] Main memory 2103 and program memory 2102 can also be designed as one memory. Alternatively, main memory 2103 and / or program memory 2102 can each be composed of a plurality of memories. Furthermore, main memory 2103 and / or program memory 2102 can also be part of processor 2101.

[0247] Processor 2101 controls communication interface 2104, which is designed, for example, as a wireless and / or wired communication interface. For example, the wireless and / or wired communication interface can receive information and forward it to processor 2101 (via a wireless and / or wired communication path) and / or can receive information from processor 2101 and transmit it (via a wireless and / or wired communication path).

[0248] An example of a wireless communication interface is a wireless network adapter. For example, the wireless communication interface comprises, in addition to an antenna, at least one transmitter circuit and one receiver circuit, or one transceiver circuit. Examples of wireless communication interfaces include GSM, UMTS, LTE and / or 5G interfaces and / or WLAN and / or Bluetooth interfaces. As mentioned above, the GSM, UMTS, LTE and 5G specifications are maintained and developed by the 3rd Generation Partnership Project (3GPP) and are currently available on the Internet at www.3gpp.com. For example, WLAN is specified in the IEEE 802. 11 Bluetooth specifications are currently available on the Internet at www.bluetooth.org.

[0249] An example of a wired communication interface is a wired network adapter. For example, the wired communication interface comprises at least one transmitter circuit and one receiver circuit, or one transceiver circuit. An example of a wired communication interface is an Ethernet interface. Ethernet is specified, inter alia, in the IEEE 802.3 series of standards.

[0250] For example, the communication interface 2104 is configured for receiving information and / or transmitting information (e.g. from another device).

[0251] Further, the processor 2101 controls a user interface 2105 which is configured for acquiring information in the form of user input (e.g. keyboard or mouse input and / or voice input and / or gestures) and / or outputting information in the form of (for example) audible and / or visual user output (e.g. voice output and / or text output). The user interface can be (for example) a keyboard, a mouse, a camera, a screen, a touch-sensitive screen, a loudspeaker, and / or a microphone.

[0252] The components 2101 to 2105 of the device 2100 are communicatively and / or operatively connected between each other, for example, by one or more bus systems (e.g. one or more serial and / or parallel bus connections).

[0253] In further examples, the device 2100 can comprise further components than the components 2101 to 2105.

[0254] The device 2100 can be understood as a server or an end device (e.g. a stationary device like a personal computer or a mobile device like a telephone, a smartphone, a tablet, a laptop), to name a few non-limiting examples. In other examples, the device 2100 can likewise be a component of any electronic device like a chip, a circuit on a chip, or a board plug-in, to name a few non-limiting examples.

[0255] Figure 22 An exemplary embodiment of a storage medium is shown. The storage medium can be (for example) a magnetic, an electronic, an optical, and / or other type of storage medium. The storage medium can be (for example) part of the processor (e.g. the processor 2101 of the device 2100 in Figure 21 An exemplary embodiment of a storage medium is shown. The storage medium can be (for example) a magnetic, an electronic, an optical, and / or other type of storage medium. The storage medium can be (for example) part of the processor (e.g. the processor 2101 of the device 2100 in

[0256] As an exemplary embodiment of the present disclosure, the following embodiments are disclosed: Exemplary embodiment 1 : A computer-implemented method comprising: - obtaining a respective impurity level of at least one impurity for each of a plurality of raw materials; - obtaining a recipe for a cell culture medium, wherein the recipe shows a ratio of the plurality of raw materials in the cell culture medium; and - determining, based at least in part on the recipe and the respective impurity levels, a total impurity level of the at least one impurity in the cell culture medium.

[0257] Example Embodiment 2: The method according to embodiment 1, further comprising: - determining, based at least in part on the recipe and the respective impurity levels of the at least one other impurity for each of the plurality of raw materials, at least one other total impurity level of the at least one other impurity in the cell culture medium.

[0258] Example Embodiment 3: The method according to embodiment 1 or embodiment 2, further comprising: - determining at least one impurity level of the respective impurity levels for each of the plurality of raw materials; and / or - outputting information indicative of the total impurity level of the at least one impurity in the cell culture medium and / or the at least one other total impurity level of the at least one other impurity in the cell culture medium.

[0259] Example Embodiment 4: The method according to any one of embodiments 1 to 3, wherein the respective impurity levels for each of the plurality of raw materials refer to respective batches of the plurality of raw materials.

[0260] Example Embodiment 5: The method according to any one of embodiments 1 to 4, further comprising: - determining, based at least in part on the total impurity level in the cell culture medium, the recipe, and the respective impurity levels, a respective contribution of one or more of the plurality of raw materials to the total impurity level and / or the at least one other total impurity level in the cell culture medium.

[0261] Example Embodiment 6: The method according to embodiment 5, further comprising: - outputting information indicative of the one or more of the plurality of raw materials having a contribution to the total impurity level and / or the at least one other total impurity level greater than a predefined threshold.

[0262] Example Embodiment 7: The method according to any one of embodiments 1 to 6, further comprising: - determining whether the total impurity level and / or the at least one other total impurity level meets at least one predefined criterion.

[0263] Example Embodiment 8: The method according to embodiment 7, wherein determining whether the total impurity level and / or the at least one other total impurity meets at least one predefined criterion further comprises at least one of: - determining whether the total impurity level and / or the at least one other total impurity level in the cell culture medium is / are below a predefined upper limit; and / or - determining whether the total impurity level and / or the at least one other total impurity level in the cell culture medium is / are above a predefined lower limit.

[0264] Example Embodiment 9: The method according to any one of embodiments 7 and 8, further comprising: - determining that a functional impurity needs to be added to the cell culture medium if it is determined that the total impurity level of the functional impurity in the cell culture medium does not meet the at least one predefined criterion; in particular, - outputting information indicating that a functional impurity needs to be added to the cell culture medium.

[0265] Example Embodiment 10: The method according to any one of embodiments 7 to 9, wherein it is determined whether the total impurity level and / or the at least one other total impurity level meets the at least one predefined criterion based at least partly on an upstream process model and / or a downstream process model of the cell culture medium.

[0266] Example Embodiment 11: A computer-implemented method, comprising: - obtaining a respective impurity level of at least one impurity for each of a plurality of raw materials; - obtaining a recipe for a cell culture medium, wherein the recipe shows a proportion of the plurality of raw materials in the cell culture medium; and - determining a total impurity level of at least one impurity in the cell culture medium based at least partly on the recipe and the respective impurity levels, wherein it is determined whether the total impurity level and / or the at least one other total impurity level meets the at least one predefined criterion based at least partly on an upstream process model and / or a downstream process model of the cell culture medium.

[0267] Example Embodiment 12: The method according to any one of embodiments 10 and 11, wherein the upstream process model comprises an impurity level of at least one impurity in at least one supplement added to the cell culture medium in the upstream process, and wherein the impurity level of the at least one impurity in the at least one supplement is added to the total impurity level before determining whether the total impurity level meets the at least one predefined criterion.

[0268] Example Embodiment 13: The method according to any one of embodiments 10 to 12, wherein the downstream process model comprises at least one factor representing a change of the total impurity level in the downstream process, and wherein it is determined whether the total impurity level meets the at least one predefined criterion based at least partly on the at least one factor.

[0269] Example Embodiment 14: The method according to any one of embodiments 5 to 13, further comprising: - outputting information indicating that the plurality of raw materials and the recipe are suitable for use in manufacturing the cell culture medium, if it is determined that the total impurity level and / or the further total impurity level fulfills at least one predefined criterion.

[0270] Example Embodiment 15: The method according to any one of embodiments 1 to 14, further comprising: - determining an allowed range of the impurity level of at least one impurity of at least one raw material of the plurality of raw materials based at least partly on the recipe, the respective impurity level of each raw material of the plurality of raw materials, and the at least one predefined criterion.

[0271] Example Embodiment 16: A computer-implemented method, comprising: - obtaining a respective impurity level of at least one impurity of each raw material of a plurality of raw materials; - obtaining a recipe of a cell culture medium, wherein the recipe shows a proportion of the plurality of raw materials in the cell culture medium; - determining a total impurity level of the at least one impurity in the cell culture medium based at least partly on the recipe and the respective impurity level; and - determining an allowed range of the impurity level of at least one impurity of at least one raw material of the plurality of raw materials based at least partly on the recipe, the respective impurity level of each raw material of the plurality of raw materials, and the at least one predefined criterion, such that the total impurity level of the at least one impurity in the cell culture medium still fulfills the predefined criterion for the total impurity when the respective impurity level of the at least one impurity of the at least one raw material varies within the allowed range.

[0272] Example Embodiment 17: The method according to any one of embodiments 15 and 16, wherein determining the allowed range of the impurity level of at least one impurity of at least one raw material of the plurality of raw materials comprises: - calculating a probability that the total impurity level of the at least one impurity in the cell culture medium does not fulfill the at least one predefined criterion based at least partly on an initial range of the impurity level of the at least one impurity of the at least one raw material; and - determining the allowed range of the impurity level based on the calculated probability and the initial range of the impurity level, in particular by using an optimization algorithm.

[0273] Example Embodiment 18: The method according to any one of embodiments 1 to 17, wherein the impurity level of at least one impurity in at least one of the plurality of raw materials is determined individually within an allowed range based on the individual impurity level of at least one impurity of those raw materials which are determined to contribute more than a predetermined threshold to the total impurity level of at least one impurity in the cell culture medium.

[0274] Exemplary embodiment 19: The method according to any one of embodiments 15 to 18, wherein the method further comprises: - outputting information indicative of the allowed range of the impurity level of at least one raw material individually.

[0275] Exemplary embodiment 20: The method according to any one of embodiments 1 to 19, further comprising: - determining a deviation of the total impurity level and / or at least one other total impurity level in the cell culture medium from the respective reference total impurity level in the cell culture medium.

[0276] Exemplary embodiment 21: The method according to embodiment 20, further comprising: - determining the individual contribution of one or more of the plurality of raw materials to the determined deviation based at least in part on the total impurity level and / or at least one other total impurity level in the cell culture medium, the formulation and the individual impurity level of each raw material; and - determining one or more raw materials having a contribution to the determined deviation greater than a predefined threshold.

[0277] Exemplary embodiment 22: The method according to any one of embodiments 1 to 21, further comprising: - determining a preferred batch of at least one of the plurality of raw materials individually, wherein the deviation of the total impurity level and / or at least one other total impurity level in the cell culture medium from the respective reference total impurity level in the cell culture medium is reduced, in particular minimized, by using an optimization algorithm based at least in part on the impurity level of at least one impurity in the preferred batch of at least one raw material.

[0278] Exemplary embodiment 23: The method according to embodiment 22, wherein the preferred batch of each of the one or more raw materials having a contribution to the determined deviation greater than a predefined threshold is determined.

[0279] Exemplary embodiment 24: The method according to any one of embodiments 1 to 23, further comprising: - determining a respective equivalent raw material for at least one of the plurality of raw materials, wherein the deviation of the total impurity level and / or the at least one other total impurity level from the respective reference total impurity level in the cell culture medium is reduced, in particular minimized, at least partially based on an impurity level of at least one impurity in the respective equivalent raw material.

[0280] Example embodiment 25: The method according to any one of embodiments 1 to 24, wherein a plurality of recipes for each cell culture medium is obtained, wherein each recipe of the plurality of recipes shows a proportion of the plurality of raw materials in each cell culture medium; and wherein the method further comprises at least one of: - determining a respective allowed range of the impurity level of the at least one impurity based on the plurality of recipes; and / or - determining a respective preferred batch of the at least one raw material from the plurality of recipes.

[0281] Example embodiment 26: The method according to any one of embodiments 1 to 25, wherein the at least one impurity and / or the at least one other impurity is one of: - a toxic impurity; or - a functional impurity; or - an organic compound; or - an inorganic compound; or - a biological load; or - an endotoxin.

[0282] Example embodiment 27: The method according to any one of embodiments 1 to 26, wherein the cell culture medium is in liquid form, and the recipe of the cell culture medium further indicates a hydration procedure for the plurality of raw materials.

[0283] Example embodiment 28: A computer program comprising instructions which, when the program is executed by a computer, cause the computer to carry out the method of any one of embodiments 1 to 27.

[0284] Example embodiment 29: An apparatus comprising means for carrying out the method of any one of embodiments 1 to 27.

[0285] It should be understood that the embodiments disclosed herein are merely exemplary and that any feature described in relation to a specific embodiment can be used in combination with any other feature described in relation to that or another specific embodiment, and / or in combination with any other feature not mentioned in relation to that or another specific embodiment. It should also be understood that any feature described in relation to an exemplary embodiment in a specific category can also be used in a corresponding manner in relation to an exemplary embodiment in any other category.

[0286] In the present disclosure, the expression "A, or B, or C, or a combination thereof" or "at least one of A, B, and / or C" can be understood as not being exhaustive, including at least: (i) A, or (ii) B, or (iii) C, or (iv) A and B, or (v) A and C, or (vi) B and C, or (vii) A, B, and C.

Claims

1. A computer-implemented method (300) comprising: - obtaining (310) an individual impurity level of at least one impurity for each of a plurality of raw materials (110; 120; 130); - obtaining (320) a recipe (140) for a cell culture medium, wherein the recipe (140) shows a proportion of the plurality of raw materials (110; 120; 130) in the cell culture medium; - determining (330), based at least in part on the recipe (140) and the individual impurity levels, a total impurity level of the at least one impurity in the cell culture medium; and - determining (410), based at least in part on the recipe (140), the individual impurity levels of the at least one impurity for each of the plurality of raw materials (110; 120; 130), and at least one predefined criterion for the total impurity level, an allowed range for the individual impurity level of the at least one impurity in at least one of the plurality of raw materials (110; 120; 130) such that the total impurity level of the at least one impurity in the cell culture medium still meets the predefined criterion for the total impurity when the individual impurity level of the at least one impurity in the at least one of the plurality of raw materials varies within the allowed range.

2. The method (300) according to claim 1, further comprising: - determining, based at least in part on the recipe (140) and the individual impurity levels of at least one other impurity for each of the plurality of raw materials (110; 120; 130), at least one other total impurity level of the at least one other impurity in the cell culture medium. 120； 3. The method (300) according to claim 1 or claim 2, further comprising: - determining at least one of the individual impurity levels of the plurality of raw materials (110; 120; 130); and / or - outputting information indicative of the total impurity level of the at least one impurity in the cell culture medium and / or the at least one other total impurity level of the at least one other impurity in the cell culture medium.

4. The method (300) according to any one of claims 1 to 3, further comprising: - determining, based at least in part on the total impurity level in the cell culture medium, the recipe (140) and the individual impurity levels, a respective contribution of one or more of the plurality of raw materials (110; 120; 130) to the total impurity level and / or the at least one other total impurity level of the cell culture medium; and - outputting information indicative of the one or more of the plurality of raw materials having a contribution to the total impurity level and / or the at least one other total impurity level that is greater than a predefined threshold.

5. The method (300) according to any one of claims 1 to 4, further comprising: - determining (340) whether the total impurity level and / or the at least one other total impurity level meets at least one predefined criterion. ​ ​ ​ 6. The method (300) of claim 5, wherein it is determined whether the total impurity level and / or the at least one further total impurity level fulfills at least one predefined criterion based at least partly on an upstream process model (150) and / or a downstream process model (160) of the cell culture medium.

7. The method (300) of claim 6, wherein the upstream process model (150) comprises an impurity level of at least one impurity in at least one supplement added to the cell culture medium in an upstream process, and wherein the impurity level of the at least one impurity in the at least one supplement is added to the total impurity level before it is determined whether the total impurity level fulfills the at least one predefined criterion.

8. The method (300) of any one of claims 6 and 7, wherein the downstream process model (160) comprises at least one factor representing a change of a total impurity level in a downstream process, and wherein it is determined whether the total impurity level fulfills at least one predefined criterion based at least partly on the at least one factor.

9. The method (300; 400) of any one of claims 1 to 8, wherein determining (410) the respective allowed range of the impurity level of at least one impurity in at least one of the plurality of raw materials (110; 120; 130) comprises: - calculating a probability that the total impurity level of the at least one impurity in the cell culture medium does not fulfill at least one predefined criterion based at least partly on an initial range of the impurity level of the at least one impurity in the at least one raw material; and - determining the respective allowed range of the impurity level based on the calculated probability and the initial range of the impurity level, in particular by using an optimization algorithm.

10. The method (300; 400) of any one of claims 1 to 9, wherein the respective allowed range of the impurity level of at least one impurity in at least one of the plurality of raw materials (110; 120; 130) is determined (410) based on the respective impurity level of at least one impurity of those raw materials which are determined to contribute more than a predefined threshold to the total impurity level of at least one impurity in the cell culture medium.

11. The method (300) of any one of claims 1 to 10, further comprising: - determining a deviation of the total impurity level and / or the at least one further total impurity level in the cell culture medium from a respective reference total impurity level in the cell culture medium; - determining a respective contribution of one or more of the plurality of raw materials (110; 120; 130) to the determined deviation based at least partly on the total impurity level and / or the at least one further total impurity in the cell culture medium, the recipe (140), and the respective impurity level of each raw material; and - determining one or more raw materials which contribute more than a predefined threshold to the determined deviation.

12. The method (300; 800) of any one of claims 1 to 11, further comprising: ​ - determining (810) a preferred batch of each of the at least one raw material, wherein a deviation of the total impurity level and / or the at least one other total impurity level of the cell culture medium from a respective reference total impurity level in the cell culture medium is reduced, in particular minimized, by using an optimization algorithm (900), based at least in part on an impurity level of at least one impurity in the preferred batch of the at least one raw material.

13. The method (300; 800) of claim 12, wherein the preferred batch of each of the one or more raw materials is determined for which the contribution to the determined deviation is greater than a predefined threshold.

14. The method (300) of any one of claims 1 to 13, wherein a plurality of recipes (140) for each cell culture medium is obtained, wherein each recipe (140) of the plurality of recipes shows a proportion of a plurality of raw materials (110; 120; 130) in each cell culture medium, and wherein the method further comprises at least one of: - determining a respective allowed range of impurity levels of the at least one impurity based on the plurality of recipes (140); and / or - determining a preferred batch of each of the at least one raw material based on the plurality of recipes (140).

15. The method (300) of any one of claims 1 to 14, wherein the at least one impurity and / or the at least one other impurity is one of: - a toxic impurity; or - a functional impurity; or - an organic compound; or - an inorganic compound; or - a biological load; or - an endotoxin.

16. A computer program comprising instructions which, when the computer program is executed by a computer, cause the computer to carry out the method (300) according to any one of claims 1 to 15.

17. An apparatus (2100) comprising means (2101; 2102; 2103) for carrying out the method (300) according to any one of claims 1 to 15.

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