CN115386610A - Culture medium for regulating antibody glycoform and method and application for regulating antibody glycoform - Google Patents
Culture medium for regulating antibody glycoform and method and application for regulating antibody glycoform Download PDFInfo
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- 238000000034 method Methods 0.000 title claims abstract description 27
- 239000001963 growth medium Substances 0.000 title claims abstract description 20
- 230000001105 regulatory effect Effects 0.000 title abstract description 23
- 230000013595 glycosylation Effects 0.000 claims abstract description 21
- 238000006206 glycosylation reaction Methods 0.000 claims abstract description 20
- 210000004027 cell Anatomy 0.000 claims description 23
- 229930182830 galactose Natural products 0.000 claims description 12
- 239000002609 medium Substances 0.000 claims description 10
- SHZGCJCMOBCMKK-UHFFFAOYSA-N D-mannomethylose Natural products CC1OC(O)C(O)C(O)C1O SHZGCJCMOBCMKK-UHFFFAOYSA-N 0.000 claims description 9
- WQZGKKKJIJFFOK-QTVWNMPRSA-N D-mannopyranose Chemical compound OC[C@H]1OC(O)[C@@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-QTVWNMPRSA-N 0.000 claims description 9
- PNNNRSAQSRJVSB-SLPGGIOYSA-N Fucose Natural products C[C@H](O)[C@@H](O)[C@H](O)[C@H](O)C=O PNNNRSAQSRJVSB-SLPGGIOYSA-N 0.000 claims description 9
- SHZGCJCMOBCMKK-DHVFOXMCSA-N L-fucopyranose Chemical compound C[C@@H]1OC(O)[C@@H](O)[C@H](O)[C@@H]1O SHZGCJCMOBCMKK-DHVFOXMCSA-N 0.000 claims description 9
- WQZGKKKJIJFFOK-PHYPRBDBSA-N alpha-D-galactose Chemical compound OC[C@H]1O[C@H](O)[C@H](O)[C@@H](O)[C@H]1O WQZGKKKJIJFFOK-PHYPRBDBSA-N 0.000 claims description 9
- 210000004978 chinese hamster ovary cell Anatomy 0.000 claims description 9
- FRHBOQMZUOWXQL-UHFFFAOYSA-L ammonium ferric citrate Chemical compound [NH4+].[Fe+3].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O FRHBOQMZUOWXQL-UHFFFAOYSA-L 0.000 claims description 8
- 239000004313 iron ammonium citrate Substances 0.000 claims description 8
- 235000000011 iron ammonium citrate Nutrition 0.000 claims description 8
- PJUIMOJAAPLTRJ-UHFFFAOYSA-N monothioglycerol Chemical compound OCC(O)CS PJUIMOJAAPLTRJ-UHFFFAOYSA-N 0.000 claims description 8
- 229940035024 thioglycerol Drugs 0.000 claims description 8
- NWONKYPBYAMBJT-UHFFFAOYSA-L zinc sulfate Chemical compound [Zn+2].[O-]S([O-])(=O)=O NWONKYPBYAMBJT-UHFFFAOYSA-L 0.000 claims description 8
- 229910000368 zinc sulfate Inorganic materials 0.000 claims description 8
- 229960001763 zinc sulfate Drugs 0.000 claims description 8
- 102000004169 proteins and genes Human genes 0.000 claims description 7
- 108090000623 proteins and genes Proteins 0.000 claims description 7
- 229960004642 ferric ammonium citrate Drugs 0.000 claims description 3
- 241000238631 Hexapoda Species 0.000 claims description 2
- 238000012258 culturing Methods 0.000 claims description 2
- 210000004408 hybridoma Anatomy 0.000 claims description 2
- 210000004962 mammalian cell Anatomy 0.000 claims description 2
- 230000004048 modification Effects 0.000 description 13
- 238000012986 modification Methods 0.000 description 13
- 238000004113 cell culture Methods 0.000 description 11
- 230000000052 comparative effect Effects 0.000 description 7
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 6
- 238000005119 centrifugation Methods 0.000 description 6
- 239000003814 drug Substances 0.000 description 6
- 239000008103 glucose Substances 0.000 description 6
- 238000011081 inoculation Methods 0.000 description 6
- 239000007788 liquid Substances 0.000 description 6
- 238000001742 protein purification Methods 0.000 description 5
- 102000003886 Glycoproteins Human genes 0.000 description 4
- 108090000288 Glycoproteins Proteins 0.000 description 4
- ISAKRJDGNUQOIC-UHFFFAOYSA-N Uracil Chemical compound O=C1C=CNC(=O)N1 ISAKRJDGNUQOIC-UHFFFAOYSA-N 0.000 description 4
- 229940079593 drug Drugs 0.000 description 4
- 239000007640 basal medium Substances 0.000 description 3
- 230000015572 biosynthetic process Effects 0.000 description 3
- 230000003204 osmotic effect Effects 0.000 description 3
- 238000011160 research Methods 0.000 description 3
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 2
- SQVRNKJHWKZAKO-UHFFFAOYSA-N beta-N-Acetyl-D-neuraminic acid Natural products CC(=O)NC1C(O)CC(O)(C(O)=O)OC1C(O)C(O)CO SQVRNKJHWKZAKO-UHFFFAOYSA-N 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 230000006870 function Effects 0.000 description 2
- 229910052760 oxygen Inorganic materials 0.000 description 2
- 239000001301 oxygen Substances 0.000 description 2
- 229920001184 polypeptide Polymers 0.000 description 2
- 102000004196 processed proteins & peptides Human genes 0.000 description 2
- 108090000765 processed proteins & peptides Proteins 0.000 description 2
- 238000012216 screening Methods 0.000 description 2
- SQVRNKJHWKZAKO-OQPLDHBCSA-N sialic acid Chemical compound CC(=O)N[C@@H]1[C@@H](O)C[C@@](O)(C(O)=O)OC1[C@H](O)[C@H](O)CO SQVRNKJHWKZAKO-OQPLDHBCSA-N 0.000 description 2
- MFBOGIVSZKQAPD-UHFFFAOYSA-M sodium butyrate Chemical compound [Na+].CCCC([O-])=O MFBOGIVSZKQAPD-UHFFFAOYSA-M 0.000 description 2
- 229940035893 uracil Drugs 0.000 description 2
- AGBQKNBQESQNJD-SSDOTTSWSA-N (R)-lipoic acid Chemical compound OC(=O)CCCC[C@@H]1CCSS1 AGBQKNBQESQNJD-SSDOTTSWSA-N 0.000 description 1
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 description 1
- 102000035195 Peptidases Human genes 0.000 description 1
- 108091005804 Peptidases Proteins 0.000 description 1
- 239000004365 Protease Substances 0.000 description 1
- AGBQKNBQESQNJD-UHFFFAOYSA-N alpha-Lipoic acid Natural products OC(=O)CCCCC1CCSS1 AGBQKNBQESQNJD-UHFFFAOYSA-N 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 229960000074 biopharmaceutical Drugs 0.000 description 1
- 239000001110 calcium chloride Substances 0.000 description 1
- 229910001628 calcium chloride Inorganic materials 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 230000001276 controlling effect Effects 0.000 description 1
- 238000001816 cooling Methods 0.000 description 1
- 229910000365 copper sulfate Inorganic materials 0.000 description 1
- ARUVKPQLZAKDPS-UHFFFAOYSA-L copper(II) sulfate Chemical compound [Cu+2].[O-][S+2]([O-])([O-])[O-] ARUVKPQLZAKDPS-UHFFFAOYSA-L 0.000 description 1
- 230000002596 correlated effect Effects 0.000 description 1
- 230000000875 corresponding effect Effects 0.000 description 1
- 230000022811 deglycosylation Effects 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 230000008030 elimination Effects 0.000 description 1
- 238000003379 elimination reaction Methods 0.000 description 1
- 230000009088 enzymatic function Effects 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 239000012526 feed medium Substances 0.000 description 1
- 150000004676 glycans Chemical class 0.000 description 1
- 238000003306 harvesting Methods 0.000 description 1
- 230000007062 hydrolysis Effects 0.000 description 1
- 238000006460 hydrolysis reaction Methods 0.000 description 1
- 230000001900 immune effect Effects 0.000 description 1
- 230000005847 immunogenicity Effects 0.000 description 1
- 235000019136 lipoic acid Nutrition 0.000 description 1
- 229920002521 macromolecule Polymers 0.000 description 1
- 238000005621 mannosylation reaction Methods 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 229940125645 monoclonal antibody drug Drugs 0.000 description 1
- 150000002772 monosaccharides Chemical class 0.000 description 1
- 239000002547 new drug Substances 0.000 description 1
- 229920001282 polysaccharide Polymers 0.000 description 1
- 239000005017 polysaccharide Substances 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 238000012827 research and development Methods 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 230000001502 supplementing effect Effects 0.000 description 1
- 238000011191 terminal modification Methods 0.000 description 1
- 229960002663 thioctic acid Drugs 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P21/00—Preparation of peptides or proteins
- C12P21/005—Glycopeptides, glycoproteins
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
- C07K16/08—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from viruses
- C07K16/10—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from viruses from RNA viruses
Abstract
The invention relates to the technical field of biology, and particularly provides a culture medium for regulating antibody glycoform, a method for regulating antibody glycoform and application thereof. The method provided by the invention is simple to operate, can realize the regulation and control of the antibody glycoform by adjusting the concentration of the components in the culture medium, controls the glycoform within a standard range, ensures the consistency and stability of antibody glycosylation, and meets the product quality requirement.
Description
Technical Field
The invention relates to the technical field of biology, in particular to a culture medium for regulating antibody glycoform, a method for regulating antibody glycoform and application
Background
With the rapid development of the biological pharmaceutical technology, the expression and preparation technology of the monoclonal antibody is greatly improved, and simultaneously, higher requirements on the aspects of research and development of the monoclonal antibody medicine, safe medication and the like are provided. The monoclonal antibody is a protein macromolecule, the structure, activity and function of the monoclonal antibody are greatly influenced by post-translational glycosylation modification, a sugar chain formed by glycosylation modification can maintain the spatial conformation of the antibody and stabilize the structure of the antibody, and research shows that the stability of the antibody to heat is greatly reduced after deglycosylation; the sugar chain structure also protects the antibody from hydrolysis by certain proteases; the special glycoform in the sugar chain is also closely related to the immune effect function, and the glycoform is not biosynthesized in a human body, so that the glycoform possibly has certain immunogenicity and can accelerate the plasma elimination of the monoclonal antibody. In general, glycosylation modification plays an important role in maintaining the structure of monoclonal antibodies, and certain types of glycosylation modification and different degrees of glycosylation modification affect the stability, safety and effectiveness of proteins. In the process of producing monoclonal antibody drugs, glycosylation modification can be influenced by various process parameters, heterogeneity is easy to generate, and the glycosylation consistency and stability of biological drugs, especially biological similar drugs must be regulated and controlled.
One of the methods for regulating glycosylation biosynthesis in the biopharmaceutical industry today is to genetically modify host cells such that specific enzymatic functions of glycoprotein biosynthesis are inhibited or deleted. The second method for regulating glycosylation biosynthesis is to optimally regulate glycosylation by a cell culture process, and comprises the steps of improving the formula of a glycoform regulator, optimizing cell culture process parameters (pH, dissolved oxygen, osmotic pressure, temperature and the like), screening a basic feed medium and the like. Sugar type regulator Mn commonly used at present 2+ Sodium butyrate, uracil, galactose and the like can meet the requirements of glycosylation engineering by screening the concentration of the sodium butyrate, the uracil, the galactose and the like. Various parameter variables influencing the glycosylation of antibody protein in the cell culture process comprise pH, which may influence terminal modifications such as antibody galactose, fucose, sialic acid and the like, and researches show that the pH is in the range of 6.8-7.2, the galactosylation modification level is reduced along with the increase of the pH, and researches also show that the pH is in the range of 6.9-7.1, the G1F + G2F glycoform is increased along with the increase of the pH, and the glycoform is not changed greatly beyond the pH range; the influence of dissolved oxygen on antibody glycoforms is mostly concentrated on the change of galactose residues; when NaCl is used for regulating osmotic pressure, the glycosylation modification of the high mannose of the monoclonal antibody is gradually increased along with the increase of the osmotic pressure; the expression level of the antibody can be improved by cooling in the process of engineering cell culture, and the influence on the glycoform of the antibody is researched, so that the low-temperature culture strategy obviously reduces the sialic acid and galactose levels; the above-mentioned technological parameters are explained for sugarThe influence of the type is not only due to a single parameter, but also needs to comprehensively consider other factors, so that specific problem specific analysis is needed for optimizing the cell culture process parameters. Patent application CN104974979A discloses a method for regulating glycosylation modification, in which method I, cell strains expressing antibodies are cultured by regulating the ratio of a basic culture medium and a supplemented culture medium, so that glycosylation modification of the antibodies is regulated; in the method II, the cell strain expressing the antibody is cultured by feeding a supplemented medium, so that the glycosylation modification of the antibody is regulated, but the method is complicated to operate.
Therefore, a method which is simpler to operate and can precisely adjust the glycoform of the antibody by adjusting the components in the medium is currently lacking.
Disclosure of Invention
Aiming at the problems in the prior art, the invention provides a culture medium for regulating antibody glycoform, a method for regulating antibody glycoform and application thereof.
In order to realize the purpose, the technical scheme adopted by the invention is as follows:
the invention provides a culture medium for regulating antibody glycoform, which comprises thioglycerol, ferric ammonium citrate and zinc sulfate.
The concentration of the thioglycerol is 1ng/L-5g/L; the concentration of the ammonium ferric citrate is 10ng/L-5g/L; the concentration of the zinc sulfate is 10ng/L-4g/L.
The invention also provides a method for regulating antibody glycoform, which uses the culture medium to culture cell lines expressing the antibody, thereby regulating the antibody glycoform.
Preferably, the glycoforms include mannose, galactose and fucose.
Preferably, the antibody is mozuri Wei Kangti (the second antibody in chinese patent CN 104603149B).
Preferably, the cell lines include mammalian cells, insect cells and hybridoma cells for expression of proteins, antibodies.
Further preferably, the cell line comprises chinese hamster ovary cells (CHO cells).
Preferably, the concentration of the thioglycerol is 5mg/L; the concentration of the ammonium ferric citrate is 0.5g/L; the concentration of the zinc sulfate is 0.1g/L.
Preferably, the conditions of the culture are: the temperature is 25-38.5 ℃, the culture period is 8-15 days, the rotating speed of a shaking table is 50-180rpm 2 The concentration is 3-10%.
Further preferably, the culture conditions are: the temperature is 36.5 ℃, the culture period is 15 days, the rotation speed of the shaking table is 140rpm 2 The concentration was 5%.
The invention also provides application of the method in regulating and controlling the glycosylation level of galactose, fucose and/or mannose.
The sugar in the present invention means a monosaccharide or a polysaccharide;
the glycoprotein described in the present invention refers to a protein or polypeptide having a sugar chain, and thus, where a sugar chain is attached to a polypeptide, a glycoprotein is defined;
the glycoform in the invention refers to a sugar chain connected to the FC terminal ASN297 of glycoprotein;
the mannose content described in the present invention comprises Man5;
CM116 described in this invention is thioglycerol; CM021 is ammonium ferric citrate; CM091 is zinc sulfate.
Compared with the prior art, the invention has the following beneficial effects:
the culture medium for regulating the antibody glycoform comprises thioglycerol, ferric ammonium citrate and zinc sulfate components, and the glycoform can be controlled within a standard range through regulation of the components; the method for adjusting the antibody glycoform can accurately adjust the components in the culture medium, control the glycoform within a standard range under the condition of not influencing the expression quantity of the antibody, ensure the glycosylation consistency and stability of the biological medicine, is simple, is easy to control the glycoform, improves the glycosylation modification type, and can be widely applied to the production of antibody imitation drugs and new drugs.
Detailed Description
The present invention will be further illustrated in detail with reference to the following specific examples, which are not intended to limit the present invention but are merely illustrative thereof. The experimental methods used in the following examples are not specifically described, and the materials, reagents and the like used in the following examples are generally commercially available under the usual conditions without specific descriptions.
The basic media in the examples and comparative examples were: the basal medium of patent application 202010676735.1.
Example 1
Adding 5mg/L CM116, 0.5g/L CM021 and 0.1g/L CM091 into the basic culture medium according to the proportion of (0.8 +/-0.1) multiplied by 10 6 cell/mL inoculation Density CHO cells were inoculated into 125mL shake flasks containing medium at a shake flask liquid loading of 30mL, a cell culture temperature of 36.5 ℃, a shaker rotation speed of 140rpm, CO 2 The concentration is 5%, the sugar concentration is measured in the period, glucose is added in time according to the sugar consumption condition, cells are harvested after the 15 th day of culture, antibody protein purification is carried out after centrifugation, and the sugar type of the purified antibody is detected.
Example 2
Adding CM116 at a concentration of 1ng/L, CM021 at a concentration of 10ng/L and CM091 at a concentration of 10ng/L to the basal medium according to a formula of (0.8 +/-0.1) multiplied by 10 6 cell/mL inoculation Density CHO cells were inoculated into 125mL shake flasks containing medium at a shake flask liquid loading of 30mL, a cell culture temperature of 36.5 ℃, a shaker rotation speed of 140rpm, CO 2 The concentration is 5%, the sugar concentration is measured in the period, glucose is added in time according to the sugar consumption condition, cells are harvested after the 15 th day of culture, antibody protein purification is carried out after centrifugation, and the sugar type of the purified antibody is detected.
Example 3
Adding 5g/L CM116, 5g/L CM021 and 4g/L CM091 into the basic culture medium according to the proportion of (0.8 +/-0.1) multiplied by 10 6 cell/mL inoculation Density CHO cells were inoculated into 125mL shake flasks containing medium at a shake flask liquid loading of 30mL, a cell culture temperature of 36.5 ℃, a shaker rotation speed of 140rpm, CO 2 At a concentration ofAnd 5%, measuring the sugar concentration in the period, timely supplementing glucose according to the sugar consumption condition, culturing until 15 days to harvest cells, purifying antibody protein after centrifugation, and detecting the sugar type of the purified antibody.
Comparative example 1
Adding 8g/L CM116, 8g/L CM021 and 7g/L CM091 into the basic culture medium according to the proportion of (0.8 +/-0.1) multiplied by 10 6 cells/mL inoculation Density CHO cells were inoculated into 125mL shake flasks containing medium at a shake flask liquid volume of 30mL, a cell culture temperature of 36.5 ℃, a shaker rotation speed of 140rpm, CO 2 The concentration is 5%, the sugar concentration is measured in the period, glucose is added in time according to the sugar consumption condition, cells are harvested after the 15 th day of culture, antibody protein purification is carried out after centrifugation, and the sugar type of the purified antibody is detected.
Comparative example 2
Adding 0.5ng/L CM116, 5ng/L CM021 and 5ng/L CM091 to the basic culture medium according to the proportion of (0.8 +/-0.1) multiplied by 10 6 cell/mL inoculation Density CHO cells were inoculated into 125mL shake flasks containing medium at a shake flask liquid loading of 30mL, a cell culture temperature of 36.5 ℃, a shaker rotation speed of 140rpm, CO 2 The concentration is 5%, the sugar concentration is measured in the period, glucose is added in time according to the sugar consumption condition, cells are harvested after the 15 th day of culture, antibody protein purification is carried out after centrifugation, and the sugar type of the purified antibody is detected.
Comparative example 3
Adding calcium chloride with concentration of 0.5g/L, thioctic acid with concentration of 5mg/L and copper sulfate with concentration of 0.1g/L into basal medium according to (0.8 + -0.1) × 10 6 cell/mL inoculation Density CHO cells were inoculated into 125mL shake flasks containing medium at a shake flask liquid loading of 30mL, a cell culture temperature of 36.5 ℃, a shaker rotation speed of 140rpm, CO 2 The concentration is 5%, the sugar concentration is measured in the period, glucose is added in time according to the sugar consumption condition, cells are harvested after the 15 th day of culture, antibody protein purification is carried out after centrifugation, and the sugar type of the purified antibody is detected.
The results are shown in Table 1
TABLE 1
| Galactose (%) | Fucose (%) | Mannose (%) | |
| Blank group | 6.7％ | 95.8％ | 4.2％ |
| Example 1 | 10.3％** | 91.5％** | 10.7％** |
| Example 2 | 17.4％** | 87.1％** | 14.8％** |
| Example 3 | 29.8％** | 93.9％** | 5.4％** |
| Comparative example 1 | 7.2％ | 95.8％ | 3.9％ |
| Comparative example 2 | 6.8％ | 9.51％ | 4.6％ |
| Comparative example 3 | 7.1％ | 95.3％ | 4.3％ |
Note: p <0.01 as compared to blank
The culture medium in the embodiment has a very significant influence on the contents of galactose, fucose and mannose of the cultured antibody, and the components CM116 and CM021 in the culture medium are positively correlated with the galactose content; CM116 and CM091 are inversely related to fucose content and positively related to mannose content, and the corresponding galactose, fucose or mannosylation level can be controlled by adjusting the component concentration of the above three.
Finally, it should be noted that the above-mentioned contents are only used for illustrating the technical solutions of the present invention, and not for limiting the protection scope of the present invention, and that the simple modifications or equivalent substitutions of the technical solutions of the present invention by those of ordinary skill in the art can be made without departing from the spirit and scope of the technical solutions of the present invention.
Claims (10)
1. A medium for modulating antibody glycoform, comprising: comprises thioglycerol, ferric ammonium citrate and zinc sulfate.
2. The culture medium according to claim 1, wherein: the concentration of the thioglycerol is 1ng/L-5g/L; the concentration of the ammonium ferric citrate is 10ng/L-5g/L; the concentration of the zinc sulfate is 10ng/L-4g/L.
3. A method of modulating glycoforms of an antibody, comprising: culturing a cell line expressing an antibody using the medium of claim 1 or 2, thereby modulating the antibody glycoform.
4. The method of claim 3, wherein: the glycoform comprises mannose, galactose and fucose; the antibody comprises mozuri Wei Kangti.
5. The method of claim 3, wherein: the cell strain comprises mammalian cells, insect cells and hybridoma cells for expressing proteins and antibodies.
6. The method of claim 5, wherein: the cell strain comprises Chinese hamster ovary cells.
7. The method of claim 3, wherein: the concentration of the thioglycerol is 5mg/L; the concentration of the ammonium ferric citrate is 0.5g/L; the concentration of the zinc sulfate is 0.1g/L.
8. The method of claim 3, wherein: the culture conditions are as follows: the temperature is 25-38.5 ℃, the culture period is 8-15 days, the rotating speed of a shaking table is 50-180rpm 2 The concentration is 3-10%.
9. The method of claim 8, wherein: the culture conditions are as follows: the temperature is 36.5 ℃, the culture period is 15 days, the rotating speed of a shaking table is 140rpm 2 The concentration was 5%.
10. Use of a culture medium according to claim 1 or 2 or a method according to any one of claims 3-9 for modulating the level of galactose, fucose and/or mannose glycosylation.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN116590371A (en) * | 2023-07-13 | 2023-08-15 | 智享生物(苏州)有限公司 | Cell culture method for reducing high mannose type antibody in Chinese hamster ovary cells |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN116590371A (en) * | 2023-07-13 | 2023-08-15 | 智享生物(苏州)有限公司 | Cell culture method for reducing high mannose type antibody in Chinese hamster ovary cells |
| CN116590371B (en) * | 2023-07-13 | 2023-10-17 | 智享生物(苏州)有限公司 | Cell culture method for reducing high mannose type antibody in Chinese hamster ovary cells |
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