Development of serum-free media for a recombinant CHO cell line producing recombinant antibody

Kim, Do Yun; Lee, Joon Chul; Chang, Ho Nam; Oh, Deok-Kun

doi:10.1016/j.enzmictec.2005.11.047

Cited by 27 publications

(20 citation statements)

References 15 publications

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“…However, cell viability and antibody concentration slightly decreased when the concentration of dextran sulfate exceeded 50 mg/l. It is likely that excessive supplementation of dextran sulfate was toxic to the cells, which is in agreement with previous reports (Kim et al 2006). Moreover, the toxicity of dextran sulfate the increase of the molecular weight (Dee et al 1997).…”

Section: Discussionsupporting

confidence: 81%

“…Putrescine is one of the polyamines which are cellular constituents shown to be essential for a variety of cell processes related to growth and differentiation, such as DNA replication, transcription and translation (Janne et al 2004). It has been widely utilized as an additive in the development in serum-free media and various literature references have reported its prominent effect on cell growth and protein expression (Jeon et al 2010;Kim et al 2006). In addition, there have been reports that polyamines enhance IgM productivity of the hybridoma cells in spite of their growth suppression activity (Sugahara et al 2008).…”

Section: Discussionmentioning

confidence: 99%

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Rational development of a serum-free medium and fed-batch process for a GS-CHO cell line expressing recombinant antibody

et al. 2012

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A serum-free medium (CHO-SFM) together with a fed-batch process was developed for the cultivation of a recombinant GS-CHO cell line producing TNFR-Fc. According to the metabolic characteristics of GS-CHO cell, a basal medium was prepared by supplementing DMEM:F12:RPMI1640 (2:1:1) with amino acids, insulin, transferrin, Pluronic F68 and some other ingredients. Statistical optimization approaches based on Plackett-Burman and central composite designs were then adopted to identify additional positive determinants and determine their optimal concentrations, which resulted in the final CHO-SFM medium formulations. The maximum antibody titer reached was 90.95 mg/l in the developed CHO-SFM, which was a 18 % and 10 fold higher than that observed in the commercial EX-CELL TM 302 medium (76.95 mg/l) and basal medium (8.28 mg/l), respectively. Subsequently, a reliable, reproducible and robust fed-batch strategy was designed according to the offline measurement of glucose, giving a final antibody yield of 378 mg/l, which was a threefold improvement over that in conventional batch culture (122 mg/l) using CHO-SFM. In conclusion, the use of design of experiment (DoE) method facilitated the development of CHO-SFM medium and fed-batch process for the production of recombinant antibody using GS-CHO cells.

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Section: Discussionsupporting

confidence: 81%

Section: Discussionmentioning

confidence: 99%

Rational development of a serum-free medium and fed-batch process for a GS-CHO cell line expressing recombinant antibody

et al. 2012

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“…This result suggests that cells need a substantial time during the early stage of the adaptation process to achieve optimal growth [17]. Overall, adequate growth and viability of rCHO cells in serum-free conditions were obtained after an adaptation process of 130 days, which is in accordance with the results of previous studies on the adaptation of CHO cells producing recombinant antibodies [18,19]. The PDL of the rCHO cell culture was relatively constant during the adaptation process with some variations in full serum-free conditions that may be attributed to the observed cell aggregation.…”

Section: Adaptation Of Rcho Cells To Serum-free Conditions and Rhepo supporting

confidence: 90%

Adaptation of CHO cells in serum‐free conditions for erythropoietin production: Application of EVOP technique for process optimization

Jukić

Bubenik²,

Pavlović

et al. 2016

Biotech and App Biochem

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Mammalian cell cultures are the preferred expression systems for the production of biopharmaceuticals requiring posttranslational processing. Usually, cell cultures are cultivated in medium supplemented with serum, which supports cell proliferation, viability, and productivity. However, due to scientific and regulatory concerns, serum-free conditions are required in recombinant protein production. Cell lines that are intended for commercial recombinant protein production have to adapt to serum- or protein-free conditions early in their development. This is a labor- and time-consuming process because of the specific cell requirements related to their adaptation in new microenvironment. In the present study, a Chinese hamster ovary (CHO) cell line producing glycosylated recombinant human erythropoietin (rhEPO) was adapted for growth and rhEPO production in serum- and protein-free conditions. The physiology, growth parameters, and morphology of the CHO cells and rhEPO biosynthesis and structure were closely monitored during the adaptation process to avoid unwanted selection of cell subpopulations. The results showed that the CHO cells were successfully adapted to suspension growth and rhEPO production in the protein-free conditions and that the structure of rhEPO remained nearly unchanged. In addition, during rhEPO production in the protein-free suspension conditions, the agitation rate seem to be significant for optimal process performance in contrast to the initial cell concentration, evaluated through evolutionary operation method.

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“…It is well known that the production of recombinant proteins in serum-free conditions is both desirable and less effective (Kim et al 2006). To keep the advantages of safe protein production in animal protein free media one can use low-cost hydrolysates such as yeast, soy, wheat gluten, or rice hydrolysates as medium additives (Sung et al 2004).…”

Section: Discussionmentioning

confidence: 99%

Production of anti TNF-α antibodies in eukaryotic cells using different combinations of vectors carrying heavy and light chains

et al. 2014

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Tumor necrosis factor-a (TNF-a) plays a key role in rheumatoid arthritis and some other autoimmune diseases. Therapy with anti-TNF-a recombinant antibodies (Ab) appears to be highly effective. Production of new hyper-producing eukaryotic cell lines can decrease the treatment cost, which currently is very high. However, due to the complexity of protein transcription, translation, processing, and secretion in mammalian cells, the stages at which antibody expression is affected are still poorly determined. The aim of this work was to compare the productivity of two cell lines developed in CHO DG44 cells, deficient in dihydrofolate reductase, transfected with vectors carrying either heavy (H) or light (L) chains of chimeric antibody under different combinations of selective elements. Both H and L chains were cloned either in pOptiVEC or pcDNA3.3 vectors and different combinations were used to produce HL and LH cell lines. We have shown that Ab production has been low and comparable between HL and LH cells until selection on methotrexate (MTX) when LH but not HL cells have responded with 3.5 times increased productivity. Flow cytometry analysis has demonstrated that intracellular concentration of full size Abs in LH cells was 5.6 times higher than in HL ones due to higher amount of H chain synthesis. No differences in viability between HL and LH cells have been found. We have concluded that the expression of H chain in the pOptiVEC vector, which is responsible for MTX resistance, has led to the suppression of H chain synthesis and limitation in full Ab assembly.

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Development of serum-free media for a recombinant CHO cell line producing recombinant antibody

Cited by 27 publications

References 15 publications

Rational development of a serum-free medium and fed-batch process for a GS-CHO cell line expressing recombinant antibody

Rational development of a serum-free medium and fed-batch process for a GS-CHO cell line expressing recombinant antibody

Adaptation of CHO cells in serum‐free conditions for erythropoietin production: Application of EVOP technique for process optimization

Production of anti TNF-α antibodies in eukaryotic cells using different combinations of vectors carrying heavy and light chains

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