Industry view on the relative importance of “clonality” of biopharmaceutical-producing cell lines

Frye, Christopher C.; Deshpande, Rohini; Estes, Scott; Francissen, Kathy; Joly, John C.; Lubiniecki, Anthony S.; Munro, Trent P.; Russell, Reb J.; Wang, Tongtong; Anderson, Karin

doi:10.1016/j.biologicals.2016.01.001

Cited by 86 publications

(105 citation statements)

References 24 publications

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“…However, a comprehensive genome and epigenome characterization of a CHO-K1 host cell line adapted to growth in three different media showed high variation in genome sequence both as a result of media adaptation and under constant culture conditions over time (Feichtinger et al, 2016). Based on these observations and as previously stressed by Frye et al (2016), absolute genetic homogeneity in a cell culture does not seem achievable because of the genomic plasticity inherent in immortalized mammalian cell lines.…”

Section: Clonal Variationmentioning

confidence: 95%

“…This original cell line has led to a multitude of commercially available and proprietary CHO cell lines (Wurm, 2013). Being an immortalized cell line, the genome of CHO cells is inherently unstable (Frye et al, 2016). Moreover, dihydrofylate reductase deficiency (DHFR) in the widely used DXB11 and DG44 cell lines was achieved by subjecting cells to radiation-and chemicalmediated mutagenesis (Urlaub et al, 1983;Urlaub and Chasin, 1980).…”

Section: Cho Host Cell Linesmentioning

confidence: 99%

“…In fact, CHO cells are known for being able to adapt to changes in process conditions, which has been exploited in the industry to generate clonally derived cell lines with enhanced manufacturing capabilities (Frye et al, 2016). Moreover, the majority of cells in host cell lines seem to be intrinsically incapable of high production of MAb, and universally competent cells are likely relatively rare cases (O'Callaghan et al, 2010).…”

Section: Clonal Variationmentioning

confidence: 99%

See 2 more Smart Citations

Improving the secretory capacity of Chinese hamster ovary cells by ectopic expression of effector genes: Lessons learned and future directions

Hansen

Pristovšek

Kildegaard

et al. 2017

Biotechnology Advances

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Section: Clonal Variationmentioning

confidence: 95%

Section: Cho Host Cell Linesmentioning

confidence: 99%

Section: Clonal Variationmentioning

confidence: 99%

See 1 more Smart Citation

Improving the secretory capacity of Chinese hamster ovary cells by ectopic expression of effector genes: Lessons learned and future directions

Hansen

Pristovšek

Kildegaard

et al. 2017

Biotechnology Advances

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“…In terms of protein analytics, there is much higher sensitivity and selectivity for product characterization and low level SV detection (Feeney et al, 2013;Wan et al, 1999). These advancements have revealed more on product heterogeneity and allow for better understanding of the impacts of heterogeneity on protein efficacy and safety (Frye et al, 2016;Pilbrough et al, 2009).…”

Section: Introductionmentioning

confidence: 99%

Utilization of sequence variants as biomarkers to analyze population dynamics in cloned cell lines

Lin

Beal

DeGruttola

et al. 2017

Biotech & Bioengineering

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The inherent nature of cloned CHO cell lines includes the presence of genetic and phenotypic drift that leads to heterogeneous populations. The genetic heterogeneity exhibited by these cells can be exploited to understand the population dynamics of cloned cell lines. Understanding the interplay between heterogeneity, cell culture conditions, and population dynamics will allow for critical assessment of overarching cell line development methods and strategies in terms of population and monoclonality. Sequence variants (SVs) are protein isoforms of the gene-of-interest that contain unintended amino acid substitutions, extensions, or truncations that may contribute to heterogeneity. In this case, SVs are unique sequences in the genome of the integrated transgene that can be used as biomarkers to understand the heterogeneity of a monoclonal cell line and how production process conditions can impact population dynamics. In this study, orthogonal genetic and analytical methods were used to examine the variability of SV levels in four different SV-containing cell lines under varied culture conditions and generational ages. Culture conditions tested had little to no impact on SV levels. However, generational age studies showed two distinct trends: stability of SV levels out to approximately 100 generations in cell lines with higher level SVs (>10%) and a progressive decrease of SV levels as the cells age to approximately 100 generations in cell lines with lower level SVs (<10%). The results suggest that the four SV-containing cell lines fall into two distinct population models; SVs present in the whole population of cells and SVs present in only a sub-population of cells. The data presented here are one of the first studies to not only analyze and compare SV levels in both genetic and protein material but also to utilize SVs as biomarkers to probe distinct populations of cloned cell lines. Biotechnol. Bioeng. 2017;114: 1744-1752. © 2017 Wiley Periodicals, Inc.

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“…Such methods allow automated screening of a large number of clones to identify the small subset of high producers. Since the present industry regulatory authorities demand assurance of 'clonality' of the production cell line, this first screening step may need to be appropriately validated to assure clonality 55 . Once the initial high throughput screening step discards the low producers, clones are typically screened at higher culture volumes like, in shake flasks or TubeSpin® tubes.…”

Section: Strategies For High and Medium Throughput Clone Screeningmentioning

confidence: 99%

Cell Culture Processes for Biopharmaceutical Manufacturing

Gadgil

2017

Current Science

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Recombinant proteins manufactured using animal cell culture processes comprise a significant fraction of biopharmaceuticals. With the expiry of patents on this class of therapeutics, there is also a significant interest in manufacture of biosimilar versions of such therapeutics. This article provides a birds-eye view of upstream process development for animal cell culture processes, with a focus on advances pertinent to the development of processes for biosimilars.

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Industry view on the relative importance of “clonality” of biopharmaceutical-producing cell lines

Cited by 86 publications

References 24 publications

Improving the secretory capacity of Chinese hamster ovary cells by ectopic expression of effector genes: Lessons learned and future directions

Improving the secretory capacity of Chinese hamster ovary cells by ectopic expression of effector genes: Lessons learned and future directions

Utilization of sequence variants as biomarkers to analyze population dynamics in cloned cell lines

Cell Culture Processes for Biopharmaceutical Manufacturing

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