Better and faster: improvements and optimization for mammalian recombinant protein production

Almo, Steven C.; Love, J.

doi:10.1016/j.sbi.2014.03.006

Cited by 64 publications

(46 citation statements)

References 51 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…In particular, the New York Structural Genomics Research Consortium (NYSGRC) has developed a system for large-scale production of proteins from eukaryotic and anaerobic organisms [108,109]. All details of protein production are harvested into a sophisticated database that is integrated with structural and functional databases into one universal system.…”

Section: Resultsmentioning

confidence: 99%

The impact of structural genomics: the first quindecennial

Grabowski

Niedzialkowska

Zimmerman

et al. 2016

J Struct Funct Genomics

View full text Add to dashboard Cite

The period 2000–2015 brought the advent of high-throughput approaches to protein structure determination. With the overall funding on the order of $2 billion (in 2010 dollars), the structural genomics (SG) consortia established worldwide have developed pipelines for target selection, protein production, sample preparation, crystallization, and structure determination by X-ray crystallography and NMR. These efforts resulted in the determination of over 13,500 protein structures, mostly from unique protein families, and increased the structural coverage of the expanding protein universe. SG programs contributed over 4,400 publications to the scientific literature. The NIH-funded Protein Structure Initiatives (PSI) alone have produced over 2,000 scientific publications, which to date have attracted more than 93,000 citations. Software and database developments that were necessary to handle high-throughput structure determination workflows have led to structures of better quality and improved integrity of the associated data. Organized and accessible data have a positive impact on the reproducibility of scientific experiments. Most of the experimental data generated by the SG centers are freely available to the community and has been utilized by scientists in various fields of research. SG projects have created, improved, streamlined, and validated many protocols for protein production and crystallization, data collection, and functional analysis, significantly benefiting biological and biomedical research.

show abstract

Section: Resultsmentioning

confidence: 99%

The impact of structural genomics: the first quindecennial

Grabowski

Niedzialkowska

Zimmerman

et al. 2016

J Struct Funct Genomics

View full text Add to dashboard Cite

show abstract

“…Productivity has been improved over the last 20 years due to the identification and selection of specific host cells, vector developments, cell culture conditions and downstream processing [1,2]. Among mammalian cells, CHO cells are the predominant host for commercial production of therapeutic proteins because of well-characterised platform technologies that allow for transfection, amplification and selection of high-producing clones [3,4].…”

Section: Introductionmentioning

confidence: 99%

Assessment of UCOE on Recombinant EPO Production and Expression Stability in Amplified Chinese Hamster Ovary Cells

Betts

Dickson

2015

Mol Biotechnol

View full text Add to dashboard Cite

CHO cells are the most frequently used host for commercial production of therapeutic proteins, and DHFR-mediated gene amplification is extensively applied to generate cell lines with increased protein production. However, decreased protein productivity is observed unpredictably during the time required for scale-up with consequences for yield, time, finance and regulatory approval. In this study, we have examined the interaction between Ubiquitous Chromatin Opening Elements (UCOE) and DHFR-linked amplification in relation to cell expression stability. In summary, the inclusion of UCOE elements generated cells that (1) achieved higher cell densities and exhibited increased production of recombinant mRNA per cell and protein yield, (2) allowed isolation of greater numbers of high-producing clones, (3) resulted in greater mRNA recovery per recombinant gene copy, (4) retained stable mRNA and protein expression after amplification provided Methotrexate (MTX) was present (but not in the absence of MTX when instability was observed) and (5) conferred copy number-dependent expression to linked transgene, suggesting that they are resistant to positional gene-silencing effects. It was concluded that the inclusion of UCOEs within expression constructs offers significant advantages for certainty of cell line generation (and the number of recovered clones for more detailed characterisation/optimisation) and that UCOEs are compatible with DHFR amplification protocols. The data suggested that enhanced cell line recovery by transcriptional enhancement of selection markers, such as DHFR, could be achieved.

show abstract

“…Th eir suitability has led to safe production of numerous recombinant therapeutics for humans, like mAb production (Birch andRacher 2006, Kelley 2009). Other advantages of these cells are their fast growth properties, their ease of transfection, their adaptability to serum-free medium, and their ability to perform complex post-translational modifi cations required for biological activity and compatibility with humans (Almo and Love 2014, Omasa et al 2010, Rita Costa et al 2010). However, the growth of mammalian cells is slower than that of the microbial host systems, the cultivation processes are longer, and the engineering selection and construction of a mammalian cell line is more complex.…”

Section: Introductionmentioning

confidence: 99%

Advancement in recombinant protein production using a marine oxygen carrier to enhance oxygen transfer in a CHO-S cell line

Pape

Bossard

Dutheil

et al. 2015

Artificial Cells, Nanomedicine, and Biotechnology

View full text Add to dashboard Cite

Recombinant proteins, particularly proteins used as therapeutics, are widely expressed for bioprocessing manufacturing processes. Mammalian cell lines represent the major host cells for bioproduction, according to their capacities of post-translational modifications and folding of secreted proteins. Many parameters can affect cell productivity, especially the rate of oxygen transfer. Dissolved oxygen, in high or low proportions, is a crucial parameter which can affect cell viability and thus productivity. HEMARINA has developed a new technology, commercially proposed as HEMOXCell(®), to improve cell culture at a large production scale. HEMOXCell(®) is a marine oxygen carrier having properties of high oxygen sensitivity, to be used as an oxygen additive during cell culture manufacturing. In this study, we investigated the effects of HEMOXCell(®) on the culture of the commonly used CHO-S cell line. Two main objectives were pursued: 1) cell growth rate and viability during a batch mode process, and 2) the determination of the effect of this oxygen carrier on recombinant protein production from a CHO-transfected cell line. Our results show an increase of CHO-S cellular growth at a rate of more than four-fold in culture with HEMOXCell(®). Moreover, an extension of the growth exponential phase and high cell viability were observed. All of these benefits seem to contribute to the improvement of recombinant protein production. This work underlines several applications using this marine-type oxygen carrier for large biomanufacturing. It is a promising cell culture additive according to the increasing demand for therapeutic products such as monoclonal antibodies.

show abstract

Better and faster: improvements and optimization for mammalian recombinant protein production

Cited by 64 publications

References 51 publications

The impact of structural genomics: the first quindecennial

The impact of structural genomics: the first quindecennial

Assessment of UCOE on Recombinant EPO Production and Expression Stability in Amplified Chinese Hamster Ovary Cells

Advancement in recombinant protein production using a marine oxygen carrier to enhance oxygen transfer in a CHO-S cell line

Contact Info

Product

Resources

About