Comparison of cell propagation methods for their effect on monoclonal antibody yield in fermentors

Reuveny, Shaul; Uribe-Vélez, Daniel; Miller, L.; Macmillan, James D.

doi:10.1016/0022-1759(86)90265-6

Cited by 154 publications

(52 citation statements)

References 8 publications

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“…Interestingly, although stimulated primary T cells proliferate better at atmosO 2 (7,29,30), consistent with the findings of Mishell and Dutton (18), B cells have been shown to proliferate and produce antibodies better at physO 2 (31,32). Furthermore, many primary cell types (e.g., fibroblasts, embryonic stem cells) have been shown to grow better at physO 2 (19,33).…”

Section: Discussionsupporting

confidence: 78%

Importance of culturing primary lymphocytes at physiological oxygen levels

Atkuri

Herzenberg²,

Niemi³

et al. 2007

Proc. Natl. Acad. Sci. U.S.A.

151

132

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Although studies with primary lymphocytes are almost always conducted in CO 2 incubators maintained at atmospheric oxygen levels (atmosO 2 ; 20%), the physiological oxygen levels (physO 2 ; 5%) that cells encounter in vivo are 2-4 times lower. We show here that culturing primary T cells at atmosO 2 significantly alters the intracellular redox state (decreases intracellular glutathione, increases oxidized intracellular glutathione), whereas culturing at physO2 maintains the intracellular redox environment (intracellular glutathione/oxidized intracellular glutathione) close to its in vivo status. Furthermore, we show that CD3/CD28-induced T cell proliferation (based on proliferation index and cell yield) is higher at atmosO2 than at physO2. This apparently paradoxical finding, we suggest, may be explained by two additional findings with CD3/CD28-stimulated T cells: (i) the intracellular NO (iNO) levels are higher at physO2 than at atmosO2; and (ii) the peak expression of CD69 is significantly delayed and more sustained at physO2 that at atmosO2. Because high levels of intracellular NO and sustained CD69 tend to down-regulate T cell responses in vivo, the lower proliferative T cell responses at physO2 likely reflect the in vitro operation of the natural in vivo regulatory mechanisms. Thus, we suggest caution in culturing primary lymphocytes at atmosO2 because the requisite adaptation to nonphysiological oxygen levels may seriously skew T cell responses, particularly after several days in culture.glutathione ͉ low oxygen ͉ nitric oxide ͉ proliferation

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Section: Discussionsupporting

confidence: 78%

Importance of culturing primary lymphocytes at physiological oxygen levels

Atkuri

Herzenberg²,

Niemi³

et al. 2007

Proc. Natl. Acad. Sci. U.S.A.

151

132

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“…While a 5 µm pore filter clogged within 7 days (Reuveny et al, 1986), continuous perfusion reactors were operated without clogging for 11 days using a 25 µm pore stainless steel mesh spin-filter (Yabannavar et al, 1992) and for 49 days using a 15 µm pore filter (Deo et al, 1996).…”

Section: Spin-filtersmentioning

confidence: 99%

Mammalian cell retention devices for stirred perfusion bioreactors

Woodside¹,

Bowen²,

Piret³

1998

Current Applications of Cell Culture Engineering

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Within the spectrum of current applications for cell culture technologies, efficient large-scale mammalian cell production processes are typically carried out in stirred fed-batch or perfusion bioreactors. The specific aspects of each individual process that can be considered when determining the method of choice are presented. A major challenge for perfusion reactor design and operation is the reliability of the cell retention device. Current retention systems include cross-flow membrane filters, spin-filters, inclined settlers, continuous centrifuges and ultrasonic separators. The relative merits and limitations of these technologies for cell retention and their suitability for large-scale perfusion are discussed.

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“…Typically, Ab production in batch cultures (Table I) is limited by the depletion of an essential nutrient (31) or generation of metabolic byproducts to inhibitory levels (32). Two main approaches have evolved to maximize production in the face of these limitations: 1) the use of fed-batch culture to replenish depleted nutrients while minimizing byproduct formation or, 2) the use of perfusion culture to continuously supply fresh medium while removing spent medium and byproducts (33).…”

mentioning

confidence: 99%

Production of Engineered Antibodies in Myeloma and Hybridoma Cells

Robinson

DiStefano

Gould

et al. 1995

ACS Symposium Series

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As reviewed in this report and exemplified in a case study, recombinant antibodies (Abs) can be expressed to very high levels in lymphoid cells by optimizing gene expression and process conditions. Myeloma and hybridoma cell lines have been constructed that secrete functionally active, recombinant antibodies at up to 15 pg/c/d in non-amplified cell lines and up to 100 pg/c/d in gene-amplified cell lines. Processes have been developed that achieve final titers in fed-batch culture of 1-2 g/L, or reach volumetric productivities in perfusion culture of 0.2-0.6 g/L/d. In the specific example of the production of a humanized anti-CD18 Ab, h1B4, amplified NS0 cell lines were developed that secreted Ab at up to 50 pg/c/d. Periodic addition of concentrated nutrient solutions in fed-batch culture allowed production of the Ab to 1.8 g/L. In perfusion culture, titers of up to 0.5 g/L were obtained with maximum volumetric (i.e. reactor volume) productivities in excess of 0.6 g/L/d. Production of Engineered AntibodiesBecause of their extraordinary molecular recognition capabilities and exquisite specificity, Abs have been considered for many potential therapeutic interventions, including (as discussed below) the short-circuiting of the inflammatory reaction. Abs can be engineered via recombinant DNA technology to optimize their therapeutic efficacy or expression levels (as reviewed in 1-2). For example, the non-binding domains of non-human Abs can be replaced by the corresponding human sequences to minimize potential human antimouse antibody (KAMA) responses (3-4); the isotype of the constant region can be changed to modulate effector function interactions (4-5); the binding domains can be modified to enhance affinity or specificity (6); or Abs of the appropriate specificity can be selected from cloned E. Coli or phage combinatorial libraries (7)(8). Such Abs can be efficiently expressed with proper post-translational modification in a number of mammalian cell systems, the most common being Chinese Hamster Ovary cells (9-11) and, as 1 Current address: DuPont Merck Pharmaceutical Co.,

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Comparison of cell propagation methods for their effect on monoclonal antibody yield in fermentors

Cited by 154 publications

References 8 publications

Importance of culturing primary lymphocytes at physiological oxygen levels

Importance of culturing primary lymphocytes at physiological oxygen levels

Mammalian cell retention devices for stirred perfusion bioreactors

Production of Engineered Antibodies in Myeloma and Hybridoma Cells

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