CHO cells in biotechnology for production of recombinant proteins: current state and further potential

Kim, Jee Yon; Kim, Yeon‐Gu; Lee, Gyun Min

doi:10.1007/s00253-011-3758-5

Cited by 632 publications

(510 citation statements)

References 127 publications

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“…Research in this direction has been conducted to efficiently optimize the medium's composition, for example, by means of approaches that are based on the monitoring of changes in the concentration of the medium components and byproducts in the culture,100 as well as genomics‐ and proteomics‐based approaches 101. Through such efforts, as well as host cell modifications,102 the per‐cell production yield has increased nearly 10‐fold from 1986 to 2004 103. The composition of the various culture media that are used in biopharmaceutical manufacturing today has not been disclosed for commercial reasons, but the composition of a previously reported serum‐free culture medium that is used for CHO cells is detailed in Table 4 for reference.…”

Section: History Of Cell Culture Mediamentioning

confidence: 99%

Animal‐cell culture media: History, characteristics, and current issues

Yao

Asayama

2017

Reprod Medicine & Biology

373

278

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BackgroundCell culture technology has spread prolifically within a century, a variety of culture media has been designed. This review goes through the history, characteristics and current issues of animal‐cell culture media.MethodsA literature search was performed on PubMed and Google Scholar between 1880 and May 2016 using appropriate keywords.ResultsAt the dawn of cell culture technology, the major components of media were naturally derived products such as serum. The field then gradually shifted to the use of chemical‐based synthetic media because naturally derived ingredients have their disadvantages such as large batch‐to‐batch variation. Today, industrially important cells can be cultured in synthetic media. Nevertheless, the combinations and concentrations of the components in these media remain to be optimized. In addition, serum‐containing media are still in general use in the field of basic research. In the fields of assisted reproductive technologies and regenerative medicine, some of the medium components are naturally derived in nearly all instances.ConclusionsFurther improvements of culture media are desirable, which will certainly contribute to a reduction in the experimental variation, enhance productivity among biopharmaceuticals, improve treatment outcomes of assisted reproductive technologies, and facilitate implementation and popularization of regenerative medicine.

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Section: History Of Cell Culture Mediamentioning

confidence: 99%

Animal‐cell culture media: History, characteristics, and current issues

Yao

Asayama

2017

Reprod Medicine & Biology

373

278

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“…To increase productivity, many cellular pathways including energy metabolism, cell cycle, apoptosis and protein secretion have been extensively studied (Chen et al 2001;Kim and Lee 2002;Watanabe et al 2002;Ifandi and Al-Rubeai 2005;Seth et al 2006;Jayapal et al 2007;Kim et al 2012). Increasing cell density by either reducing apoptosis (Meents et al 2002;Figueroa et al 2007) or altering cell metabolism (Chen et al 2001) can lead to increase in product titers.…”

Section: Introductionmentioning

confidence: 99%

Dynamics of unfolded protein response in recombinant CHO cells

Prashad

Mehra

2014

Cytotechnology

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Genes in the protein secretion pathway have been targeted to increase productivity of monoclonal antibodies in Chinese hamster ovary cells. The results have been highly variable depending on the cell type and the relative amount of recombinant and target proteins. This paper presents a comprehensive study encompassing major components of the protein processing pathway in the endoplasmic reticulum (ER) to elucidate its role in recombinant cells. mRNA profiles of all major ER chaperones and unfolded protein response (UPR) pathway genes are measured at a series of time points in a high-producing cell line under the dynamic environment of a batch culture. An initial increase in IgG heavy chain mRNA levels correlates with an increase in productivity. We observe a parallel increase in the expression levels of majority of chaperones. The chaperone levels continue to increase until the end of the batch culture. In contrast, calreticulin and ERO1-L alpha, two of the lowest expressed genes exhibit transient time profiles, with peak induction on day 3. In response to increased ER stress, both the GCN2/PKR-like ER kinase and inositol-requiring enzyme-1alpha (Ire1a) signalling branch of the UPR are upregulated. Interestingly, spliced X-Box binding protein 1 (XBP1s) transcription factor from Ire1a pathway is detected from the beginning of the batch culture. Comparison with the expression levels in a low producer, show much lower induction at the end of the exponential growth phase. Thus, the unfolded protein response strongly correlates with the magnitude and timing of stress in the course of the batch culture.

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“…Current transfection strategies in cell line development are based on the random integration of the gene of interest into the host genome with a varying number of gene-copies depending on gene amplification strategies (Kim et al 2012). The cell clones obtained from these methods are quite heterogeneous and need to be screened thoroughly.…”

Section: Introductionmentioning

confidence: 99%

Transgene copy number comparison in recombinant mammalian cell lines: critical reflection of quantitative real-time PCR evaluation

et al. 2013

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Nucleic acid quantification is a relevant issue for the characterization of mammalian recombinant cell lines and also for the registration of producer clones. Quantitative real-time PCR is a powerful tool to investigate nucleic acid levels but numerous different quantification strategies exist, which sometimes lead to misinterpretation of obtained qPCR data. In contrast to absolute quantification using ampliconor plasmid standard curves, relative quantification strategies relate the gene of interest to an endogenous reference gene. The relative quantification methods also consider the amplification efficiency for the calculation of the gene copy number and thus more accurate results compared to absolute quantification methods are generated. In this study two recombinant Chinese hamster ovary cell lines were analysed for their transgene copy number using different relative quantification strategies. The individual calculation methods resulted in differences of relative gene copy numbers because efficiency calculations have strong impact on gene copy numbers. However, in context of comparing transgene copy numbers of two individual clones the influence of the calculation method is marginal. Therefore especially for the comparison of two cell lines with the identical transgene any of the relative qPCR methods was proven as powerful tool.

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CHO cells in biotechnology for production of recombinant proteins: current state and further potential

Cited by 632 publications

References 127 publications

Animal‐cell culture media: History, characteristics, and current issues

Animal‐cell culture media: History, characteristics, and current issues

Dynamics of unfolded protein response in recombinant CHO cells

Transgene copy number comparison in recombinant mammalian cell lines: critical reflection of quantitative real-time PCR evaluation

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