Optimization of a Fed‐batch Process for Production of a Recombinant Antibody

Shen

J of Chemical Tech & Biotech

2004

The achievement of high cell density and monoclonal antibody concentration depends on understanding cell metabolism and the control of cell culture. A model of cell metabolism and monoclonal antibody production in hybridoma cell culture is presented. Fed-batch culture in serum-free medium was conducted in a bioreactor with an optimized feeding strategy according to the model. In the process, nutrient concentrations, especially glucose and glutamine, were maintained at relatively constant and low levels. The formation of toxic byproducts, such as ammonia and lactate, was greatly reduced in comparison with that in non-optimized batch culture. The concentration of monoclonal antibody reached 350 mg dm −3 , which was seven times greater than in non-optimized batch culture.

Section: Introductionmentioning

confidence: 99%

Fed‐batch culture of hybridoma cells in serum‐free medium using an optimized feeding strategy

Shen

J of Chemical Tech & Biotech

2004

Biotech & Bioengineering

“…Hayter et al (1992) found that glycosylation patterns of interferon-g are dependent on the age of the cell culture while monoclonal IgG produced in ascites, hollow fibre bioreactors and static culture was shown to be differently glycosylated (Cabrera et al, 2005;Lund et al, 1993;Patel et al, 1992). Robinson et al (1994) characterised a recombinant IgG1 produced by mouse NS0 cell line during a serum-free, fed batch process. They observed that the antigen-binding characteristics of the antibody remained constant throughout the process but the carbohydrate composition changed.…”

Section: Introductionmentioning

confidence: 97%

Glycosylation of an immunoglobulin produced from a murine hybridoma cell line: The effect of culture mode and the anti‐apoptotic gene, bcl‐2

Majid

Butler

Al‐Rubeai

2006

The impact of bcl-2 over-expression on the glycosylation pattern of an antibody produced by a bcl-2 transfected hybridoma cell line (TB/C3.bcl-2) was investigated in suspension batch, continuous and high cell density culture (Flat hollow fibre, Tecnomouse system). In all culture modes bcl-2 over-expression resulted in higher cell viability. Analysis of the glycans from the IgG of batch cultures showed that >95% of the structures were neutral core fucosylated asialo biantennary oligosaccharides with variable terminal galactosylation (G0f, G1f and G2f) consistent with previous analysis of glycans from the conserved site at Asn-297 of the IgG protein. The galactosylation index (GI) was determined as an indicator of the glycan profile (=(G2 + 0.5* G1)/(G0 + G1 + G2)). GI values in control cultures were comparable to bcl-2 cultures during exponential growth (0.53) but declined toward the end of the culture when there was a loss in cell viability. Low dilution rates in chemostat culture were associated with reduced galactosylation of the IgG glycans in both cell lines. However, at the higher dilution rates the GI for IgG was consistently higher in the TB/C3.bcl-2 cultures. In the hollow fibre bioreactor the galactosylation of the IgG glycans was considerably lower than in suspension batch or continuous cultures with GI values averaging 0.38. Similar low galactosylation values have been found previously for high density cell cultures and these are consistent with the low values obtained when the dissolved oxygen level is maintained at a low value (10%) in controlled suspension cultures of hybridomas.

“…A systematic approach has been able to improve productivity, increase fed‐batch product titers, and reduce reactor volumes and cost of goods. Notably, a number of studies have reported iterative methods where basal media are supplemented with concentrated nutrient solutions5, 6 and in some cases supplements are added according to the uptake rate of the cells 7. A model based on stoichiometric nutritional demands for animal cell growth was developed8 to increase cell growth and product titers by minimizing toxic waste accumulation in the culture.…”

Section: Introductionmentioning

confidence: 99%

Host–Guest Chemistry of Aromatic‐Amide‐Linked Bis‐ and Tris‐Calix[4]pyrroles with Bis‐Carboxylates and Citrate Anion

Cafeo

Gattuso

Kohnke

et al. 2014

Chemistry A European J

A small library of polytopic receptors has been synthesized from meso-p- and meso-m-aminophenylcalix[4]pyrroles and p- or m-phthaloyl or trimesic chloride. Selected bis-carboxylates and the citrate anion, which either exhibit altered distribution profiles in cancerous tissues in comparison with healthy tissues or are metabolites of carcinogenic substances (for example, trans,trans-muconic acid from benzene exposure in humans) were tested as ligands. Varied affinities and binding modes were observed as a function of the number of calix[4]pyrroles and the topology of amide units present in each of the polytopic receptors. The structures of the 1:1 complexes derived by molecular modeling are in excellent agreement with the results of (1)H NMR complexation studies.