CN111009291A - Cell culture medium optimization method - Google Patents
Cell culture medium optimization method Download PDFInfo
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- CN111009291A CN111009291A CN201910591777.2A CN201910591777A CN111009291A CN 111009291 A CN111009291 A CN 111009291A CN 201910591777 A CN201910591777 A CN 201910591777A CN 111009291 A CN111009291 A CN 111009291A
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- G—PHYSICS
- G16—INFORMATION AND COMMUNICATION TECHNOLOGY [ICT] SPECIALLY ADAPTED FOR SPECIFIC APPLICATION FIELDS
- G16B—BIOINFORMATICS, i.e. INFORMATION AND COMMUNICATION TECHNOLOGY [ICT] SPECIALLY ADAPTED FOR GENETIC OR PROTEIN-RELATED DATA PROCESSING IN COMPUTATIONAL MOLECULAR BIOLOGY
- G16B50/00—ICT programming tools or database systems specially adapted for bioinformatics
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M3/00—Tissue, human, animal or plant cell, or virus culture apparatus
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- G—PHYSICS
- G16—INFORMATION AND COMMUNICATION TECHNOLOGY [ICT] SPECIALLY ADAPTED FOR SPECIFIC APPLICATION FIELDS
- G16C—COMPUTATIONAL CHEMISTRY; CHEMOINFORMATICS; COMPUTATIONAL MATERIALS SCIENCE
- G16C20/00—Chemoinformatics, i.e. ICT specially adapted for the handling of physicochemical or structural data of chemical particles, elements, compounds or mixtures
- G16C20/10—Analysis or design of chemical reactions, syntheses or processes
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- G—PHYSICS
- G16—INFORMATION AND COMMUNICATION TECHNOLOGY [ICT] SPECIALLY ADAPTED FOR SPECIFIC APPLICATION FIELDS
- G16C—COMPUTATIONAL CHEMISTRY; CHEMOINFORMATICS; COMPUTATIONAL MATERIALS SCIENCE
- G16C20/00—Chemoinformatics, i.e. ICT specially adapted for the handling of physicochemical or structural data of chemical particles, elements, compounds or mixtures
- G16C20/70—Machine learning, data mining or chemometrics
Abstract
The invention discloses a cell culture medium optimization method, which comprises a component titration method, a culture medium mixing method, a consumption component analysis method, a stoichiometric analysis method, a rational culture medium design method and a system culture medium development method; the method can effectively reduce the cost of the culture medium, improve the biological safety and quality stability and the compatibility of a downstream purification process, can realize the flux screening of test conditions, and greatly accelerates the development process of a cell culture process.
Description
Technical Field
The invention belongs to the technical field of culture media, and particularly relates to a cell culture medium optimization method.
Background
The culture medium is a nutrient medium prepared by combining different nutrient substances for the growth and reproduction of microorganisms, plants or animals, generally contains several substances such as carbohydrates, nitrogen-containing substances, inorganic salts, vitamins, water and the like, and is a basic substance for providing cell nutrition and promoting cell proliferation as well as a living environment for the growth and reproduction of cells.
Early animal cell culture media often contain fetal calf serum or calf serum, and the development trend of industrial culture media in recent years is turned to protein-free animal-derived culture media or even chemical component-free culture media, and in addition, antibody production enterprises have the problems of medium cost, biological safety and quality stability and compatibility of downstream purification processes in the culture medium development process, so a cell culture medium optimization method is provided.
Disclosure of Invention
The invention aims to provide a cell culture medium optimization method to solve the problems that the early animal cell culture medium proposed in the background art often contains fetal calf serum or calf serum, the development trend of the culture medium for industrial use in recent years is changed to a protein-free animal-derived or even chemical-component-free culture medium, and in addition, antibody production enterprises have the problems of culture medium cost, biological safety, quality stability and compatibility of downstream purification processes in the process of culture medium development.
In order to achieve the purpose, the invention provides the following technical scheme: a cell culture medium optimizing method comprises a component titration method, a culture medium mixing method, a consumed component analysis method, a stoichiometric analysis method, a rational culture medium design method and a system culture medium development method.
As a further illustration of the present invention, it is preferred that the component titration method is to determine the optimal concentration of a component of the medium by designing a concentration gradient of the component.
As a further illustration of the present invention, it is preferred that the media mixing method is to rapidly determine the optimal media formulation by experimental design and analysis with DOE software by mixing prototype media of known composition in different proportions.
As a further illustration of the present invention, it is preferable that the consumption component analysis method is a method of adjusting a fed-batch strategy by analyzing consumption of each component during a batch culture process, and timely supplementing an easily consumable component.
As a further illustration of the present invention, it is preferred that the chemometric analysis is a method of establishing a chemometric relationship between glucose, amino acids, vitamins and cellular biomass and recombinant protein synthesis, and energy metabolism, using a simplified mathematical model.
As a further illustration of the present invention, preferably, the rational culture medium design method comprises screening a large number of culture medium candidates and mixed formulas, performing batch culture using an initially determined basal culture medium, analyzing easily consumable components in the culture medium, designing a fed-batch culture medium and a fed-batch strategy based on the screened large number of culture medium candidates and mixed formulas, determining the optimum concentration of other additives of the fed-batch culture medium by using a component titration method, and verifying the optimized basal culture medium, fed-batch culture medium and fed-batch strategy on a mini-reactor.
As a further explanation of the present invention, it is preferable that the systematic medium development method is a method in which medium components are divided into several groups of amino acids, vitamins, nucleotides, and the like, and a group having a significant influence on cell expression is first determined, and then important component optimization is performed for the group.
As a further illustration of the present invention, it is preferred that the component titration method is applied only to optimize individual components in the culture, such as insulin, hydrolysates and growth factors.
As a further illustration of the present invention, it is preferred that the method for analyzing a consumed component is mainly used for the optimization of glucose, glutamine and amino acid components because of the limitation of detection means.
As a further illustration of the present invention, it is preferred that the stoichiometry is optimized for the initial medium, fed-batch medium, and fed-batch strategy to reduce lactic acid and ammonia production.
Compared with the prior art, the invention has the beneficial effects that: the method can effectively reduce the cost of the culture medium, improve the biological safety and quality stability and the compatibility of a downstream purification process, can realize the flux screening of test conditions, and greatly accelerates the development process of a cell culture process.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
The invention provides a technical scheme that: a cell culture medium optimizing method comprises a component titration method, a culture medium mixing method, a consumed component analysis method, a stoichiometric analysis method, a rational culture medium design method and a system culture medium development method.
In this example, it is preferred that the component titration is to determine the optimal concentration of a component of the medium by designing a concentration gradient of the component.
In this embodiment, the culture medium mixing method is preferably to perform experimental design and analysis by DOE software by mixing prototype culture media with known components in different proportions, and to quickly determine the optimal culture medium formula.
In this embodiment, preferably, the consumption component analysis method is to adjust the fed-batch strategy by analyzing the consumption of each component during the batch culture process, and timely supplementing the easily consumed component.
In this embodiment, the preferred chemometric analysis method is to use a simplified mathematical model to establish the chemometric relationship between glucose, amino acids, vitamins and cellular biomass and recombinant protein synthesis, as well as energy metabolism.
In this embodiment, preferably, the rational culture medium design method is to screen a large number of culture medium candidates and mixed formulas, perform batch culture using the initially determined basic culture medium, analyze easily consumable components in the culture medium, design a fed-batch culture medium and a fed-batch strategy based on the screened rational culture medium candidates and mixed formulas, determine the optimal concentration of other additives in the fed-batch culture medium by using a component titration method, optimize the basic culture medium, the fed-batch culture medium and the fed-batch strategy, and verify the optimal concentrations on a mini-reactor.
In this embodiment, the system medium development method is preferably a method in which medium components are divided into a plurality of groups of amino acids, vitamins, nucleotides, and the like, a group having a significant influence on cell expression is determined, and then important component optimization is performed for the group.
In this example, preferably, the component titration method is applied to optimize only individual components in the culture, such as insulin, hydrolysates and growth factors.
In this embodiment, it is preferable that the method of analyzing a consumed component is mainly used for optimizing glucose, glutamine, and amino acid components because of the limitation of the detection means.
In this embodiment, it is preferable that the stoichiometry is optimized for the initial medium, fed medium, and fed strategy to reduce the production of lactic acid and ammonia.
Although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that changes, modifications, substitutions and alterations can be made in these embodiments without departing from the principles and spirit of the invention, the scope of which is defined in the appended claims and their equivalents.
Claims (10)
1. A method of optimizing a cell culture medium, comprising: comprises a component titration method, a culture medium mixing method, a consumption component analysis method, a stoichiometric analysis method, a rational culture medium design method and a system culture medium development method.
2. A method of optimizing a cell culture medium according to claim 1, wherein: the component titration method is to determine the optimal concentration of the components of the culture medium by designing the concentration gradient of the components.
3. A method of optimizing a cell culture medium according to claim 1, wherein: the culture medium mixing method is characterized in that a prototype culture medium with known components is mixed in different proportions, and experimental design and analysis are carried out by DOE software, so that the optimal culture medium formula is quickly determined.
4. A method of optimizing a cell culture medium according to claim 1, wherein: the consumption component analysis method is characterized in that easily consumed components are supplemented in time and a fed-batch strategy is adjusted by analyzing the consumption condition of each component in the batch culture process.
5. A method of optimizing a cell culture medium according to claim 1, wherein: the chemometric analysis method is to establish the chemometric relation among glucose, amino acid, vitamin, cell biomass, recombinant protein synthesis and energy metabolism by utilizing a simplified mathematical model.
6. A method of optimizing a cell culture medium according to claim 1, wherein: the rational culture medium design method comprises the steps of screening a large number of culture medium candidates and a mixed formula, carrying out batch culture by using an initially determined basic culture medium, analyzing easily-consumed components in the culture medium, designing a fed-batch culture medium and a fed-batch strategy according to the components, determining the optimal concentration of other additives of the fed-batch culture medium by using a component titration method, optimizing the basic culture medium, the fed-batch culture medium and the feeding strategy, and carrying out verification on a mini-reactor.
7. A method of optimizing a cell culture medium according to claim 1, wherein: the system culture medium development method is characterized in that the components of a culture medium are divided into a plurality of groups of amino acid, vitamin, nucleotide and the like, firstly, the group with more obvious influence on cell expression is judged, and then, the important component optimization is carried out aiming at the group.
8. A method of optimizing a cell culture medium according to claim 1, wherein: the component titration method is only applicable to optimize individual components in culture, such as insulin, hydrolysates and growth factors.
9. A method of optimizing a cell culture medium according to claim 1, wherein: the method for analyzing the consumed components is mainly used for optimizing glucose, glutamine and amino acid components due to the limitation of detection means.
10. A method of optimizing a cell culture medium according to claim 1, wherein: the chemometric analysis method is used for optimizing an initial culture medium, a fed-batch culture medium and a fed-batch strategy, and can reduce the production of lactic acid and ammonia.
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20060148074A1 (en) * | 1996-08-30 | 2006-07-06 | Invitrogen Corporation | Serum-free mammalian cell culture medium, and uses thereof |
| CN104894055A (en) * | 2015-06-15 | 2015-09-09 | 成都金凯生物技术有限公司 | Optimized cell culture medium and cell culture method, and application of optimized cell culture medium and cell culture method to preparation of protein and antibody |
| CN106479982A (en) * | 2016-10-17 | 2017-03-08 | 深圳万乐药业有限公司 | Anti- PD 1 monoclonal antibody Cells for production culture medium and its optimization method |
-
2019
- 2019-07-02 CN CN201910591777.2A patent/CN111009291A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20060148074A1 (en) * | 1996-08-30 | 2006-07-06 | Invitrogen Corporation | Serum-free mammalian cell culture medium, and uses thereof |
| CN104894055A (en) * | 2015-06-15 | 2015-09-09 | 成都金凯生物技术有限公司 | Optimized cell culture medium and cell culture method, and application of optimized cell culture medium and cell culture method to preparation of protein and antibody |
| CN106479982A (en) * | 2016-10-17 | 2017-03-08 | 深圳万乐药业有限公司 | Anti- PD 1 monoclonal antibody Cells for production culture medium and its optimization method |
Non-Patent Citations (1)
| Title |
|---|
| 刘伯宁: "用于重组抗体生产的细胞大规模培养技术", 《中国生物工程杂志》, pages 103 - 111 * |
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