Effects of Synchronization on CD40 Expression and Antibody Production in Hybridoma Cells Stimulated with Anti-mIgG

Martín-López, Alicia; Camacho, F. Garcı́a; Contreras-Gómez, Antonio; Grima, E. Molina

doi:10.1002/bp070087t

Cited by 3 publications

(7 citation statements)

References 28 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…5a), in a similar way to that observed in a previous work for thymidine synchronization (Martín-López et al 2007a). Both the expression of CD40 and specific productivity declined abruptly after achieving the maximum value, probably provoked both by the loss of synchronization and by the decrease in the concentration of anti-mIgG in the culture medium since, as previously shown, the free anti-mIgG initially added to the culture medium was bound to IgG2a as this was secreted by cells.…”

Section: Cell Cyclesupporting

confidence: 88%

“…The results showed that some of the factors (HU synchronization, anti-mIgG treatment, a second dose of thymidine, a second dose of HU and LPS treatment) affected stronger CD40 expression (p value \ 0.05), and the increase or decrease in CD40 expression led to increased or decreased mAB productivity; but others factors (preLPS treatment and thymidine synchronization) affected stronger mAB productivity (p value \ 0.05), probably involving metabolic routes different from those of CD40. This shows a complex relationship between mAb productivity and CD40 expression that can not be represented by a simple linear regression as proposed previously (Martín-López et al 2007a). …”

Section: Cell Cyclementioning

confidence: 67%

“…The capture of secreted IgG2a by free anti-mIgG antibody added to the culture medium was determined as described previously (Martín-López et al 2007a). Statistical analyses…”

Section: Determination Of Antibody Concentrationmentioning

confidence: 99%

“…Addition of anti-mouse surface immunoglobulin G antibody (anti-mIgG) to the culture medium has been shown to increase CD40 expression as well as specific mAb productivity on 55-6 hybridoma cells (Martín-López et al 2007a, b, c). This effect was improved by increasing the proportion of cells in the G1 phase with thymidine synchronization (Martín-López et al 2007a). In addition, there was a good correlation between the increase in specific mAb productivity and the increase in CD40 expression.…”

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

Effects of hydroxyurea on monoclonal antibody production induced by anti-mIgG and LPS stimulation on murine B cell hybridomas

et al. 2010

Self Cite

View full text Add to dashboard Cite

Chemical treatment with hydroxyurea (HU) has been selected as a simple and low cost strategy to generate a cell population enriched for the G1 phase. After the chemical treatment with HU, cells were stimulated with anti-mIgG to test if the positive effects of anti-mIgG on CD40 expression and specific IgG2a production rate were improved upon a cell population with a higher percentage of cells in G1 phase at the beginning of the cell culture. In addition, other treatments assayed in this work were the cell stimulation with Lipopolysaccharide (LPS) both before and after the HU treatment. It has been observed that the use of HU under conditions able to maintain the cells in viable state (0.1 mM for 20 h), has a negative effect on CD40 expression and specific IgG2a production rate induced by anti-mIgG. The positive effect of LPS on cell stimulation induced by anti-mIgG is reduced on cells treated with HU.

show abstract

Section: Cell Cyclesupporting

confidence: 88%

Section: Cell Cyclementioning

confidence: 67%

“…The capture of secreted IgG2a by free anti-mIgG antibody added to the culture medium was determined as described previously (Martín-López et al 2007a). Statistical analyses…”

Section: Determination Of Antibody Concentrationmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Effects of hydroxyurea on monoclonal antibody production induced by anti-mIgG and LPS stimulation on murine B cell hybridomas

et al. 2010

Self Cite

View full text Add to dashboard Cite

show abstract

“…A relationship between cell cycle position, CD40 expression and mAb productivity has also been observed (Martín-López et al 2007c, 2010.…”

Section: Introductionmentioning

confidence: 78%

Co-culture of the 55-6 B cell hybridoma with the EL-4 thymoma cell. Effect on cell growth and monoclonal antibody production

et al. 2013

Self Cite

View full text Add to dashboard Cite

The cell growth and monoclonal antibody production of the 55-6 hybridoma cell co-cultured with the murine thymoma cell line EL-4 at different initial 55-6:EL-4 ratios were investigated. Both populations were seeded in co-culture without previous stimulation and therefore with low constitutive CD40 and CD40 ligand (CD154) expression levels, and in the absence of exogenous co-stimuli. Viable cell density and growth rate data seem to suggest a competition for nutrients, which is detrimental for both cells in terms of biomass production and also of growth rate for 55-6. Final concentrations of antibody and specific antibody production rates were affected by the initial 55-6:EL-4 ratio. The 4:1 ratio yielded the highest IgG2a concentration, whereas the highest specific antibody production rate was obtained at the 2:1 ratio. Changes mainly in CD154 and also in CD40 expression in cocultures could suggest cross-talk between both populations. In conclusion, different types of interactions are probably present in this co-culture system: competition for nutrients, cognate interaction and/or autocrine or paracrine interactions that influence the proliferation of both cells and the hybridoma antibody secretion. We are hereby presenting a pre-scale-up process that could speed up the optimization of largescale monoclonal antibodies production in bioreactors by emulating the in vivo cell-cell interaction between B and T cells without previous stimulation or the addition of co-stimulatory molecules.

show abstract

Model‐based identification of cell‐cycle‐dependent metabolism and putative autocrine effects in antibody producing CHO cell culture

Möller

Korte

Pörtner

et al. 2018

Biotech & Bioengineering

View full text Add to dashboard Cite

The understanding of cell-cycle-dependent population heterogeneities in mammalian cell culture and their influence on production rates is still limited. Furthermore, metabolic regulations arising from self-expressed signaling factors (autocrine/autoinhibitory factors) have been postulated in the past, but no determination of such effects have been made so far for fast-growing production Chinese hamster ovary (CHO) cells in chemically defined media. In this study, a novel approach combining near-physiological treatment of cells (including synchronization), population resolved mechanistic modeling and statistical analysis was developed to identify population inhomogeneities. Cell-cycle-dependent population dynamics and metabolic regulations due to a putative autocrine factor were examined and their impact on the metabolic rates and antibody production of near-physiologically synchronized CHO DP-12 cell cultures was determined. To achieve this, a population resolved model was extended to describe putative autocrine-dependentt and cell-cycle-related metabolic rates for glucose, glutamine, lactate, ammonia, and antibody production. The model parameters were estimated based on data of two repeated batch cultivations (three batches each), with main substrates in excess and potentially inhibiting waste products (lactate and ammonium) controlled within narrow ranges. Significant changes, due to a putative autocrine factor, were identified for lactate and ammonia formation and antibody production. The cell growth and the uptake of glucose and glutamine were only partially affected by the putative autocrine under the given conditions. The results indicate the presence of a self-expressed autocrine factor and its strong impact on the metabolism of CHO DP-12 cells. Furthermore, glucose and glutamine uptake, as well as the formation of ammonium and antibody were found to be significantly cell-cycle-dependent. The combined approach has a strong potential to improve the understanding of the interplay of population heterogeneities and signal factors in mammalian cell culture.

show abstract

Effects of Synchronization on CD40 Expression and Antibody Production in Hybridoma Cells Stimulated with Anti-mIgG

Cited by 3 publications

References 28 publications

Effects of hydroxyurea on monoclonal antibody production induced by anti-mIgG and LPS stimulation on murine B cell hybridomas

Effects of hydroxyurea on monoclonal antibody production induced by anti-mIgG and LPS stimulation on murine B cell hybridomas

Co-culture of the 55-6 B cell hybridoma with the EL-4 thymoma cell. Effect on cell growth and monoclonal antibody production

Model‐based identification of cell‐cycle‐dependent metabolism and putative autocrine effects in antibody producing CHO cell culture

Contact Info

Product

Resources

About