Porous cell culture plate
Through the combination of porous cell culture plates and fully automatic liquid workstations, different problems of post-translation modification of microbial expression proteins are solved, high-throughput screening is achieved, and the safety and effectiveness of biopharmaceuticals are improved, which is particularly suitable for the culture medium of CHO cells. optimization.
Patent Information
- Application Number
- CN202323495815.5
- Authority / Receiving Office
- CN · China
- Patent Type
- Utility models(China)
- Current Assignee / Owner
- Filing Date
- 2023-12-21
- Publication Date
- 2024-12-31
- Estimated Expiration
- 2033-12-21
AI Technical Summary
In existing biopharmaceuticals, the proteins expressed by microorganisms are different from those of humans in post-translational modification, resulting in a shortening of immunogenicity and drug half-life, affecting safety and effectiveness. The optimization of cell culture medium requires a large number of experimental screening and is inefficient.
Using porous cell culture plates combined with fully automatic liquid workstations, high-throughput screening is performed through a series of medium formulas designed by matrix, copper or aluminum metal thermal conduction plates are used to maintain uniform temperatures, and the PTFE diaphragm gastubular gas permeable membrane prevents pathogenic microorganisms from entering, realizing Automated oscillation culture and protein expression detection.
It improves the optimization experimental efficiency of cell culture medium, realizes the uniformity detection of protein expression and post-translational modification, improves the safety and effectiveness of drugs, and is particularly suitable for high-throughput screening of CHO cells.
Smart Images
Figure CN222274579U_ABST
Abstract
Description
Technical Field
[0001] The utility model relates to a biomedical research and development device, and more specifically to a multi-porous cell culture plate. Background Art
[0002] Biopharmaceuticals are a field of the pharmaceutical industry that has emerged and flourished in the 21st century. The main categories include recombinant protein drugs, monoclonal antibody drugs, vaccines, etc., which play an irreplaceable role in the treatment or prevention of many major diseases such as malignant tumors, autoimmune diseases, genetic diseases, infectious diseases, etc. It has now developed into an industry with an annual output value of hundreds of billions of US dollars, and has set off a global research and development boom. More and more research institutions and pharmaceutical companies have invested in the research and development and production of biological drugs.
[0003] Biological products such as recombinant protein drugs and monoclonal antibody drugs are currently mainly produced through bioengineering fermentation, and commonly used expression hosts include genetically recombinant bacteria, yeast, insect cells, animal cells, etc. Early recombinant proteins were mainly expressed and prepared by microorganisms such as bacteria and yeast. However, the proteins expressed by these microorganisms are different from those expressed by humans in terms of post-translational modifications, especially the differences in glycosylation modifications, which can easily cause problems such as immunogenicity and shortened half-life in vivo, thereby affecting the safety and effectiveness of the drugs. In recent years, with the development of mammalian cell culture technology, mammalian cells represented by Chinese hamster ovary (CHO) cells have become the main expression system for high-molecular-weight recombinant proteins with complex structures, especially recombinant proteins with glycosylation modifications.
[0004] The composition of cell culture medium and cell culture conditions (such as temperature, dissolved oxygen, stirring, etc.) have an important impact on the protein yield and post-translational modification expressed by CHO cells. The composition of cell culture medium is complex, including amino acids, carbon sources, vitamins, trace elements, inorganic salts, proteins and other components. In order to optimize the cell culture medium to obtain the best expression and post-translational modification, it is necessary to use DOE to design the formula ingredients and conduct a large number of experimental screenings to select the optimal combination of cell culture medium ingredients. Summary of the invention
[0005] In order to improve the experimental efficiency of cell culture medium optimization, a fully automatic liquid workstation is used in conjunction with a microplate to perform high-throughput screening of a series of culture medium formulas using a matrix design. The utility model provides a multi-porous cell culture plate, and the technical solution is as follows:
[0006] The technical solution of the utility model is:
[0007] A multi-well cell culture plate ( Figure 1), including a base 1 and a cover plate 2, the cover plate 2 is fixed to the base 1 by a groove 8. There are 2 to 1000 sample holes 3 on the base 1, and the shape of the sample hole 3 is a cone that is wide at the top and narrow at the bottom, and the bottom is circular. There is a metal heat conductive plate 4 below the sample hole 3, and the material is copper, aluminum, or an alloy containing copper or aluminum. The bottom of the metal heat conductive plate 4 is flat, and the upper interface fits the shape of the bottom of the sample hole 3. The function of the metal heat conductive plate 4 is to keep the temperature of each sample hole 3 uniform.
[0008] The outer side of the outermost sample hole 3 on the base 1 is provided with a concave water tank 5 for containing water and balancing the volatilization of the liquid in the edge holes of the cell culture plate.
[0009] A sealing strip 6 is provided between the upper edge of the base 1 and the cover plate 2 and is made of rubber or silicone for isolating gas and liquid.
[0010] The top of the cover plate 2 is provided with a bacteria-proof breathable membrane 7 made of polytetrafluoroethylene (PTFE), which is used for gas exchange between the sample hole 3 and the outside and prevents pathogenic microorganisms from entering.
[0011] An example of a method of using the utility model:
[0012] Add cell culture fluids of different formulations containing cells into the sample well 3, and the height of the liquid in the sample well 3 should not exceed 1 / 3 of the depth of the sample well 3. Then add water into the water tank 5, cover the cover plate 2 on the base 1, and place it on the shaker of the constant temperature incubator for vibration culture. After a certain period of time, take out the cell culture supernatant in each sample well 3 to detect the protein expression and post-translational modification of the protein. If combined with the fully automatic liquid workstation and its constant temperature incubation module and oscillation module, the process of adding liquid, shaking culture, and taking liquid can be automatically completed to achieve automated high-throughput screening.
[0013] Compared with common cell culture multi-well plates, the utility model has more uniform temperatures between sample wells, a large air permeability area, better air permeability, and a relatively fixed cover plate and base, which is more suitable for oscillating culture of non-adherent cells and related research, especially high-throughput screening of CHO cell culture media. BRIEF DESCRIPTION OF THE DRAWINGS
[0014] Figure 1 : A longitudinal section structure of a multi-well cell culture plate.
[0015] Figure numerals: 1-base, 2-cover plate, 3-sample hole, 4-heat conducting plate, 5-water tank, 6-sealing strip, 7-bacteria-isolating breathable membrane, 8-groove. DETAILED DESCRIPTION
[0016] Example 1: Structure of a porous cell culture plate
[0017] A multi-well cell culture plate ( Figure 1 ), including a base 1 and a cover plate 2, the cover plate 2 is fixed to the base 1 by a groove 8. There are 96 sample holes 3 on the base 1, and the shape of the sample holes 3 is a cone that is wide at the top and narrow at the bottom, and the bottom is round. There is a metal heat conductive plate 4 below the sample hole 3, which is made of copper. The bottom of the metal heat conductive plate 4 is flat, and the upper interface fits the shape of the bottom of the sample hole 3. The function of the metal heat conductive plate 4 is to keep the temperature of each sample hole 3 uniform.
[0018] The outer side of the outermost sample hole 3 on the base 1 is provided with a concave water tank 5 for containing water and balancing the volatilization of the liquid in the edge holes of the cell culture plate.
[0019] A sealing strip 6 is provided between the upper edge of the base 1 and the cover plate 2 and is made of silicone for isolating gas and liquid.
[0020] The top of the cover plate 2 is provided with a bacteria-proof breathable membrane 7 made of polytetrafluoroethylene (PTFE), which is used for gas exchange between the sample hole 3 and the outside and prevents pathogenic microorganisms from entering.
[0021] Example 2: Method for using a multi-well cell culture plate
[0022] Add cell culture fluids of different formulations containing cells into the sample well 3, and the height of the liquid in the sample well 3 should not exceed 1 / 3 of the depth of the sample well 3. Then add water into the water tank 5, cover the cover plate 2 on the base 1, and place it on the shaker of the constant temperature incubator for vibration culture. After a certain period of time, take out the cell culture supernatant in each sample well 3 to detect the protein expression and post-translational modification of the protein. If combined with the fully automatic liquid workstation and its constant temperature incubation module and oscillation module, the process of adding liquid, shaking culture, and taking liquid can be automatically completed to achieve automated high-throughput screening.
Claims
1. A multi-porous cell culture plate, comprising a base (1) and a cover plate (2), characterized in that: The cover plate (2) is fixed to the base (1) by means of a groove (8); the base (1) is provided with 2 to 1000 sample holes (3), the sample holes (3) are in the shape of a cone that is wide at the top and narrow at the bottom, and has a circular bottom; a heat conducting plate (4) is provided below the sample holes (3).
2. The multi-well cell culture plate according to claim 1, characterized in that: The outer side of the outermost sample hole (3) on the base (1) is provided with a recessed water tank (5).
3. The multi-well cell culture plate according to claim 1, characterized in that: A sealing strip (6) is provided between the upper edge of the base (1) and the cover plate (2).
4. The multi-well cell culture plate according to claim 1, characterized in that: A bacteria-isolating breathable membrane (7) is provided on the top of the cover plate (2).
5. The multi-well cell culture plate according to claim 1, characterized in that: The heat conducting plate (4) is made of copper, aluminum, or an alloy containing copper or aluminum.
6. The multi-well cell culture plate according to claim 3, characterized in that: The sealing strip (6) is made of rubber or silicone.
7. The multi-well cell culture plate according to claim 4, characterized in that: The material of the bacteria-isolating breathable membrane (7) is polytetrafluoroethylene.