WO2005095582A2 - Nouveaux supplements de milieu de culture de cellules - Google Patents

Nouveaux supplements de milieu de culture de cellules Download PDF

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Publication number
WO2005095582A2
WO2005095582A2 PCT/EP2005/002228 EP2005002228W WO2005095582A2 WO 2005095582 A2 WO2005095582 A2 WO 2005095582A2 EP 2005002228 W EP2005002228 W EP 2005002228W WO 2005095582 A2 WO2005095582 A2 WO 2005095582A2
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WO
WIPO (PCT)
Prior art keywords
cells
cell culture
culture medium
antibodies
supplement
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PCT/EP2005/002228
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English (en)
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WO2005095582A3 (fr
Inventor
Karlheinz Landauer
Günter Waxenecker
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Igeneon Krebs-Immuntherapie Forschungs-Und Entwicklungs-Ag
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Publication of WO2005095582A2 publication Critical patent/WO2005095582A2/fr
Publication of WO2005095582A3 publication Critical patent/WO2005095582A3/fr

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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
    • C12N5/0018—Culture media for cell or tissue culture
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
    • C07K16/28—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
    • C07K16/2896—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against molecules with a "CD"-designation, not provided for elsewhere
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/42—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against immunoglobulins
    • C07K16/4208—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against immunoglobulins against an idiotypic determinant on Ig
    • C07K16/4241—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against immunoglobulins against an idiotypic determinant on Ig against anti-human or anti-animal Ig
    • C07K16/4258—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against immunoglobulins against an idiotypic determinant on Ig against anti-human or anti-animal Ig against anti-receptor Ig
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2500/00—Specific components of cell culture medium
    • C12N2500/30—Organic components
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2500/00—Specific components of cell culture medium
    • C12N2500/50—Soluble polymers, e.g. polyethyleneglycol [PEG]

Definitions

  • the invention refers to a novel animal cell culture medium supplement which can increase growth rate and yield of in-vitro cultivated cells. Furthermore the invention refers to the use of the culture medium supplement for the cultivation of cells and for the increase of production of proteins expressed by the cells.
  • Cell culture media must provide the nutrients necessary to maintain and grow cells in a controlled, artificial and in-vitro environment. Characteristics and compositions of the cell culture media may vary depending on the particular cellular requirements.
  • Media formulations have been used to cultivate a number of cell types including animal, plant and bacterial cells.
  • Cells cultivated in culture media catabolize available nutrients and produce useful biological substances such as monoclonal antibodies, hormones, growth factors, viruses and the like. Such products have therapeutic applications, and, based on recombinant DNA technology, cells can be engineered to produce large quantities of these products.
  • the ability to cultivate cells in vitro is not only important for the study of cell physiology, but is also necessary for the production of useful substances which may otherwise be impossible or only obtainable by cost-effective means.
  • Culture media can be provided as liquid solutions but also as dry powder media.
  • WO 98/36051 describes dry powder cells and nutritive media powder. These powder media can be transferred into moistened powder by adding defined and controlled solvents. These solvents can be water, serum or organic solvents.
  • US 5,328,844 describes a strictly defined medium composition, COF 1769, for the cultivation of mammalian cells, containing a defined amino acid composition, antioxidants and protective agents.
  • WO 89/01028 describes serum free media for the growth of insect cells consisting of a basal medium, lipid, peptone components and protective agents to stabilize the cells from damage and death in agitated and sparged insect culture. Lots of different agents, as amino acids, trace elements, serum, vitamins, lipids and hydrolysates (e.g. soybean hydrolysate) are used in media for cell cultivation.
  • the energy source of in-vitro cultured cells is mainly glucose and L-glutamine. Glucose is degraded via glycolysis producing pyruvate and NADH.
  • TCC tri-carbonic acid cycle
  • the efficiency of glucose consumption differs from 30% up 80% depending on the cell line and culture conditions.
  • another agent has to be added to the medium to support the catabolism.
  • L-glutamine is used as further supplement in concentrations of 2 - 10 mM.
  • Glutamine is transferred to the TCC and used for anaplerotic and catabolic pathways.
  • Glutamine is degraded to glutamate and thereby producing ammonia, which is toxic.
  • the rate of degradation can be influenced by pH and ionic conditions but in all cell culture media, formation of these breakdown products often cannot be avoided (Tritsch et al., 1962, Exp.Cell Res. 28:360-364).
  • the object of the invention is solved by providing a cell culture supplement and its use as disclosed in the claims.
  • new cell culture supplements are provided which can be used either alone or in combination. It was surprisingly found that the addition of acetone either alone or in combination with ⁇ -hydroxyethylstarch (HES) can activate cells resulting in higher growth rates and/or increased specific productivities. Growth rates and/or specific productivity of the animal cells can be increased leading to higher yields of recombinant proteins expressed by the cells.
  • HES ⁇ -hydroxyethylstarch
  • the potency of the expressed proteins is equal, preferably better compared to the proteins expressed by cells cultivated in standard media.
  • In-vitro cultivated cells can metabolize acetone and increase their yield of expressed proteins up to at least 20%, preferably at least 50%, preferably at least 100%, preferably at least 150%, preferably at least 250% depending on the fermentation mode and the used cell line.
  • HES is capable to increase the yield of a bioprocess up to 20%, preferably up to
  • a mixture of HES and acetone can activate mammalian cells in order to increase the specific productivity by two fold.
  • Both substances can positively interfere, possibly via reducing the osmotic pressure, with the quality of formed proteins and enhance the metabolism of mammalian cells.
  • the invention provides the use of acetone or acetone and HES as cell culture supplements for the cultivation of cells.
  • the supplements according to the invention can easily be removed from the cells or the expressed proteins by any purification means known by the skilled person.
  • the invention also provides methods for the cultivation of cells either adherently or in suspension using a medium containing the media supplement according to the invention.
  • the invention also provides a method for the production of proteins by cultivation of host cells in a medium containing the supplement. Especially the production of antibodies, derivatives or fragments thereof is provided.
  • the invention also provides a method for obtaining an antibody or antibody derivative or fragment thereof or recombinant proteins by a cell culture which is cultivated in a medium comprising a cell culture supplement according to the invention, comprising the steps of: culturing the cells which produce antibodies or derivatives or fragments thereof in the presence of the supplement according to the invention, continuing said culture until said antibodies or derivatives or fragments accumulate, and isolating said antibodies or derivatives or fragments thereof.
  • the invention further provides a culture of cells cultivated in a cell culture medium supplemented by acetone in a concentration of 1 ⁇ M to 20mM, preferably 5 ⁇ M to 10mM, preferably 10 ⁇ M to 1mM preferably 10 ⁇ M to 100 ⁇ M and optionally together with HES, wherein HES is in a concentration of 0.01 to 10 weight%, preferably 0.1 to 5 weight%, preferably 0.2 to 2 weight%.
  • the term "animal cell culture medium" which can be supplemented by acetone and/or HES can be any medium known from the state of the art.
  • the basal medium can be a synthetic medium such as DMEM/HAM ' s F12, Medium 199, Hybridoma Express or RPMI, IMDM, Ex-Cell SP2/0, Ex-Cell 325-PF or combinations thereof, and others that are known from the literature or are commercially available.
  • the basal medium can comprise a number of ingredients, including amino acids, vitamins, organic or inorganic salts, sources of carbohydrate, each ingredient being present in an amount which supports the cultivation of a cell in vitro.
  • a nonionic surfactant such as polyethylene glycol or polypropylene glycol (PLURONIC F-61 ; PLURONIC F-68, SYNPERONIC F-68, PLURONIC F- 71 or PLURONIC F-108) can be added to the medium as a defoaming agent. These agents are well known to protect cells from the negative effect of aeration.
  • the term "animal cell culture” refers to animal cells grown in suspension in roller bottles, flasks, spinners etc.
  • Thebioprocess can be a batch-culutre, Fed-Batch- culture, a continuous or perfusion cultivation. Large scale approaches, such as bioreactors including stirred tank reactors or airlift reactors, or including cells growing attached to microcarriers in stirred bioreactors, fluidized bed reactors or airlift reactors, are also included.
  • yield can be defined as overall productivity of a bioprocess.
  • bioprocess can be defined as a method to produce a biologically derived product.
  • the term “cultivation” refers to the maintenance of the cells in vitro under conditions permissive for growth and continued viability.
  • Mammalian cells are typically cultivated in a cell incubator at a temperature between 32 and 38°C, in particular at about 37°C.
  • the optimal pH range is about 6.7 to 7.6, preferably between 6.9 to 7.2.
  • the acetone used as medium supplement can be any quality of acetone, yet "pro analysi"-grade is preferred.
  • Acetone can be added to the cell culture medium in a concentration of 1 ⁇ M to 20mM, preferably 5 ⁇ M to 10mM, preferably 10 ⁇ M to
  • the HES used as medium supplement is preferably of high purity. HES can be added to the cell culture medium in a concentration of 0.01 to 10 weight percent
  • weight% preferably 0.1 to 5 weight%, preferably 0.2 to 2 weight%.
  • Using the cell culture medium supplement in these concentrations resulted in increased cell growth and / or high yield.
  • the cell culture supplement/supplements according to the invention can be added at any time during cultivation. It can be added to the medium before inoculation of the cells or during the cultivation process.
  • the supplement can also be added continuously to keep a constant concentration of the supplement in the medium.
  • the animal cells cultivated in a medium supplemented by acetone or acetone and
  • HES can be any animal cells that can grow in vitro in synthetic media.
  • the term "cells” means the cultivation of individual cells, tissues, organs, insect cells, avian cells, mammalian cells, hybridoma cells, primary cells, continuous cell lines, stem cells and/or genetically engineered cells, such as recombinant cells expressing a heterologous protein or polypeptide.
  • the cells used according to the invention are preferably animal cells, more preferably mammalian cells. These can be for example BSC-1 cells, LLC-MK cells, CV-1 cells, CHO cells, COS cells, murine cells, human cells, HeLa cells,
  • 293 cells VERO cells, MDBK cells, MDCK cells, MDOK cells, CRFK cells, RAF cells, TCMK cells, LLC-PK cells, PK15 cells, WI-38 cells, MRC-5 cells, T-FLY cells, BHK cells, SP2/0 cells, NS0. perC6 (human retina cells) or derivatives thereof.
  • the proteins expressed by these cells can be any proteins known in the art.
  • the proteins are recombinant proteins, hormones, viruses, antibodies or derivatives or fragments thereof like. Erythropoietin, insulin or coagulation factors can be exemplary listed as these proteins.
  • antibody relates to antibodies of all possible types, in particular to polyclonal and monoclonal antibodies or antibodies produced by chemical, biochemical or gene technology methods. Methods for producing such molecules are known to the person skilled in the art.
  • the antibodies can also be used as carrier molecules for antigenic structures.
  • Antibody derivatives and fragments can be single chain antibodies and fragments thereof and miniantibodies, bispecific antibodies, diabodies, triabodies, or di-, oligo- or multimers thereof. Also included are peptidomime tics or peptides derived from antibodies e.g. they comprise one or several CDR regions. Further included are human monoclonal antibodies and peptide sequences which, based on a structure activity connection, are produced through an artificial modeling process
  • epitope defines any region of a molecule that can be recognised by specific antibody or that provoke the formation of those specific antibodies.
  • Epitopes may be either conformational epitopes or linear epitopes.
  • Preferred epitopes or antigenic structures are derived from antigens specific for epithelial tumors, and preferably frequently expressed in breast cancer, gastrointestinal, colorectal, rectal, prostate, pancreatic, and ovary and lung cancer, either being small cell lung cancer (SCLC) or non small cell lung cancer
  • SCLC small cell lung cancer
  • NSCLC Newcastle disease virus
  • the preferred epitopes especially induce humoral immune response and the formation of specific antibodies in vivo.
  • the antibodies preferably also induce T cell specific response.
  • epitopes are protein epitopes that are expressed on malignant cells of solid tumors, e.g. TAG-72, MUC1 , Folate Binding Protein A-33,
  • T cell epitope peptides or mimotopes of such T cell epitopes may be presented by the antibody according to the invention.
  • Preferred carbohydrate epitopes or antigenic structures are derived from tumor associated aberrant carbohydrate structures, such as Lewis antigens, e.g. Lewis x-, Lewis b- und Lewis y-structures, also sialylated Lewis x structures, GloboH- structures, KH1 , Tn-antigen, TF-antigen and alpha-1-3-galactosyl-epitope.
  • Lewis antigens e.g. Lewis x-, Lewis b- und Lewis y-structures, also sialylated Lewis x structures, GloboH- structures, KH1 , Tn-antigen, TF-antigen and alpha-1-3-galactosyl-epitope.
  • the preferred TAA targets or epitopes are selected from trie group of determinants derived from the group of antigens consisting of peptides or proteins, such as EpCAM, HER2-9, NCAM, CEA, TOL9-re ceptors and T cell peptides, carbohydrates, such as aberrant glycosylation patterns, Lewis Y, Sialyl-
  • Tn Tn
  • Globo H glycolipids, such as GD2, GD3 und GM2 .
  • these antibodies can be antibodies according WO 93/24647, EP 0
  • Figure 1 shows, as control, the roller bottle batch culture viable population curve, viability curve and volumetric monoclonal antibody titer curve.
  • Figure 2 Roller bottle batch culture viable population curve, viability curve and volumetric monoclonal antibody titer curve.
  • Cells E4 were propagated as free suspension in 850cm 2 roller bottles agitated with 2 rpm in 450 mL working volume. Test was performed in a double experiment; sampling was on done a one to three day basis.
  • the used medium was Hybridoma Express supplemented with 4mM L-GIutamine, 0.1 % Pluronic F68 and 0.2% HES.
  • FIG. 3 Roller bottle batch culture viable population curve, viability curve and volumetric monoclonal antibody titer curve are shown.
  • the cells E4 were propagated as free suspension in 850cm 2 roller bottles agitated with 2 rpm in 450 mL working volume. The test was performed in a double experiment; sampling was done on a one to three day basis.
  • the used medium was Hybridoma Express supplemented with 4mM L-GIutamine, 0.1 % Pluronic F68 and 10 ⁇ M acetone.
  • Figure 4 Roller bottle batch culture viable population curve, viability curve and volumetric monoclonal antibody titer curve.
  • Cells E4 were propagated as free suspension in 850cm 2 roller bottles agitated with 2 rpm in 450 mL working volume. Test was performed in a double experiment; sampling was done on a one to three day basis.
  • the used medium was Hybridoma Express supplemented with 4mM L-GIutamine, 0.1% Pluronic F68 and 10 ⁇ M acetone.
  • Figure 5 The cumulative cell numbers of living E4 hybridoma cells in serum-free medium per day. The test was performed in replicates. The line is the linear regression of all measured time points. The highest cumulative cell numbers were achieved in HES containing medium after 10 days of cultivation. Cells cultivated in the acetone medium and the HES + acetone medium performed nearly equal, while the standard medium reached only 75% of the amount. Trie slope of the standard medium is less steep than the slope of the other tested media. This indicates a lower overall growth rate of the cells.
  • Figure 6 The cumulative cell numbers of living SP2/0-311 hybridoma cells in serum-containing medium per day. In behalf of the overall growth rate (slope) no significant differences could be observed.
  • Figure 7 The specific productivity of monoclonal antibody by recombinant SP2/0 cell line determined from the linear relationship between product titer and cumulative cell-days. The cells cultivated in medium containing both new supplements reached the highest titer. The slopes of both HES containing media are the steepest suggesting HES as the best media supplement for this cell line.
  • Figure 8 The cumulative cell numbers of living CHO-E5 cells in serum-free medium per day. Regarding the overall growth rate (slope), no significant differences were observed.
  • Figure 9 The specific productivity of monoclonal antibody produced by CHO-E5 cells determined from the linear relationship between product titer and cumulative cell-days.
  • the HES supplemented medium revealed the steepest slope, followed by medium containing acetone.
  • the slopes of the control and the medium with the mixture of both novel supplements were significantly lower.
  • the cell lines used were E4, a mouse hybridoma producing anti-idiotype antibody as surrogate of the Lewis Y carbohydrate antigen and 311 a recombinant SP2/0 producing a humanized anti-Lewis Y antibody and a recombinant SP2/0 producing anti-Lewis Y antibody and a recombinant CHO-E5 producing rmAB17- 1A.
  • the basal serum-free medium for E4 was Hybridoma Express (PAA, Linz) supplemented with 4 mM L-glutamine (Sigma Aldrich Corp., St. Louis MO, USA) and 0.1 % Pluronic F68 (Sigma, St. Louis MO).
  • PAA Hybridoma Express
  • Pluronic F68 Syluronic F68
  • SP2/0 311 cell line RPMI 1640 (Gibco) supplemented with 10% Fetal Calf Seru m (Gibco) and 2 mM L-Glutamin was used.
  • the serum-free medium for the SP2/0 311 cells was a mixture of Hybridoma Express (75%) and RPMI 1640 (25%) supplemented with 6 mM L-GIutamine.
  • the CHO cells were cultivated in EX-Cell 325 PF CHO protein- free medium (JRH Biosciences, UK).
  • Cell culture was performed at 37°C in standard grade polystyrene cell culture flasks (Nunc, Roskilde, Denmark) in an incubator with T% CO 2 .
  • Cell number of the supernatant was determined by the trypan blue dye exclusion method using a hemocytometer and following standard protocol (0.2% trypan blue).
  • the growth rate was calculated by linear regression of the log of cell densities versus time of the logarithmic phase of the culture.
  • Inoculation density was 1.5 0 5 cells mL "1 .
  • the titer of serum containing cell culture supernatant was measured by ELISA following standard procedures. Photometric extinction of the developed color reaction was measured at 492 nm (reference wavelength 620 nm). All tests were performed in duplicates.
  • the next step of the development was to perform a roller bottle culture in batch mode with the same cells and supplement concentrations.
  • Table 1 Population growth and productivity batch culture parameters of cell lines (SP2/0 hybridoma, recombinant SP2/0 and CHO).
  • the recombinant CHO-E5 cell line cultivated in media containing the novel supplements displayed an increased specific productivity of 350% for HES ( Figure 9) and 220% for acetone. The combination of both supplements did not show this effect.
  • the growth rates of the cells cultured in all tested media were similar ( Figure 8). Therefore an increase of the volumetric productivity for HES (270%) and acetone (250%) could be shown.
  • Osmotic stress especially hyperosmotic stress, negatively influences the glycosylation of recombinant proteins (Schmelzer, A.E. and W.M. Miller, 2002, Biotechnology Progress 18:346-353.).
  • the novel media supplements are capable to reduce the osmolality of medium either containing serum or not, thus the glycosylation pattern can be positively influenced by simultaneously raising the available amount of energy sources.
  • Hydroxyethylstarch can be used as glucose storage.
  • the cells can degrade the polymer as needed to obtain the necessary amount of the primary energy source glucose.
  • the initial amount of glucose can be reduced and the high molecular weight of the biopolymer helps to decrease the osmolality in the medium, while maintaining high glucose levels.
  • Acetone on the other side opens a third source of energy.
  • the up-take of acetone into the cells is not known, but as it dissolves easily in fatty acids, it might freely access the cells.
  • NADPH and acetone are co-oxidized by acetone- monooxygenase producing NADP and acetol.
  • Acetol can be metabolized either via acetic acid or via pyruvic acid to acetyl-CoA.
  • the acetyl-CoA can than be degraded in the TCC to CO 2 producing ATP or it can be used to produce other metabolites, as necessary.
  • Preliminary measurements of the concentration of acetone in the media with GC-MS revealed a consumption of the supplement during batch culture.

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  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
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  • Proteomics, Peptides & Aminoacids (AREA)
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Abstract

L'invention concerne un supplément de milieu de culture de cellules animales destiné à cultiver des cellules animales, qui consiste en de l'acétone. L'invention concerne également un procédé de culture de cellules.
PCT/EP2005/002228 2004-03-05 2005-03-03 Nouveaux supplements de milieu de culture de cellules WO2005095582A2 (fr)

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ATA383/2004 2004-03-05
AT3832004A AT413700B (de) 2004-03-05 2004-03-05 Mediumzusätze für zellkultur enthaltend aceton

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WO2005095582A2 true WO2005095582A2 (fr) 2005-10-13
WO2005095582A3 WO2005095582A3 (fr) 2006-06-01

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Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN104212769A (zh) * 2014-07-14 2014-12-17 北京益生合生物科技有限公司 一种用于高产单抗的细胞培养基添加剂
EP3574985A1 (fr) 2013-12-20 2019-12-04 President And Fellows Of Harvard College Dispositifs organomimétiques et leurs procédés d'utilisation et de fabrication
CN111690057A (zh) * 2020-06-24 2020-09-22 哈尔滨元亨生物药业有限公司 一种sp2/0细胞灌注用培养基及重组犬细小病毒单克隆抗体生产方法

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5328844A (en) * 1990-05-09 1994-07-12 University Of Colorado Foundation, Inc. Culture media for mammalian cells
US20030153079A1 (en) * 2001-11-30 2003-08-14 Invitrogen Corporation Dry powder cell culture products containing lipid and methods of production thereof

Family Cites Families (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1991000341A1 (fr) * 1989-06-29 1991-01-10 Cetus Corporation Culture de cellules d'insectes par gargouillement et expression de proteines de recombinaison

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5328844A (en) * 1990-05-09 1994-07-12 University Of Colorado Foundation, Inc. Culture media for mammalian cells
US20030153079A1 (en) * 2001-11-30 2003-08-14 Invitrogen Corporation Dry powder cell culture products containing lipid and methods of production thereof

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
DATABASE BIOSIS [Online] BIOSCIENCES INFORMATION SERVICE, PHILADELPHIA, PA, US; 1992, VNUCHKOVA V A ET AL: "Using acetone in nutrient media to increase regenerant output" XP002367059 Database accession no. PREV199396131691 & DOKLADY ROSSIISKOI AKADEMII SEL'SKOKHOZYAISTVENNYKH NAUK, no. 9-10, 1992, pages 10-12, ISSN: 1068-3674 *

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP3574985A1 (fr) 2013-12-20 2019-12-04 President And Fellows Of Harvard College Dispositifs organomimétiques et leurs procédés d'utilisation et de fabrication
CN104212769A (zh) * 2014-07-14 2014-12-17 北京益生合生物科技有限公司 一种用于高产单抗的细胞培养基添加剂
CN111690057A (zh) * 2020-06-24 2020-09-22 哈尔滨元亨生物药业有限公司 一种sp2/0细胞灌注用培养基及重组犬细小病毒单克隆抗体生产方法
CN111690057B (zh) * 2020-06-24 2021-04-23 哈尔滨元亨生物药业有限公司 一种重组犬细小病毒单克隆抗体生产方法

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ATA3832004A (de) 2005-09-15
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