Biomedical Engineering and Bioengineering Commons^™

Targeted Integration Of Multiple Active Sites In Cho Genome For Rapid Generation Of Stable And High Monoclonal Antibody Producing Cell Lines, Yuansheng Yang

Cell Culture Engineering XV

Mammalian cell lines used for manufacturing recombinant therapeutic proteins need to have high productivity, long term stable production and good product quality. The process of cell line development currently used in industry is based on random integration of the plasmid vectors into genome. Due to clonal variation, hundreds to thousands of clones are screened through multiple stages of evaluation. The whole process takes at least a few months and is extremely tedious. Targeted integration-based cell line development is attractive as the plasmid vectors integrate into predetermined active sites in genomes. All targeted cell lines have same genetic background and thus …

Bridging The Gap Of Screening And Scale Up In Cho, Hybridoma, Hek293 And Other Cell Lines: Single Use Optimum Growth Flasks 30ml-5l Flasks With Transfer Caps, And Ports, Sam Ellis

Cell Culture Engineering XV

Optimum Growth Flasks^TM (patented) are a break through design to shake flasks, and the first in 100+ years. By using Optimum Growth Flasks^TM users are able to grow 15mLs-3L of Cell Culture with the same scalability. The Optimum Growth Flasks^TM have replaced disposable Fernbach Flasks and also small Wave^® Bags (5L & 10L) in the larger sizes as production vessels. The Optimum Growth Flasks ^TM also give high viability cultures with a great use of space as shown by our data in CHO, Hybridoma, HEK293 and other Cell Lines. We have made a series of Transfer …

Cho-K1 Host Cell Engineering Strategy Enabling The Establishment Of Strains Producing Higher Yields Of Recycling Antibodies, Hisahiro Tabuchi, Ikumi Murakami, Masaki Yamaguchi

Cell Culture Engineering XV

As described elsewhere (Biotechnol Bioeng 2010, 2013), our DXB11 (dhfr^–) host cell engineering strategy achieved high cell viability for a prolonged period (more than 1 month) and enhanced mAb productivity (>100 pg/cell/day) by nutritional control. Introduction of taurine transporter (TAUT) into DXB11 parent cells increased glutamine uptake and accelerated glutathione metabolism. By forcing the overexpression of TAUT, we were able to control DXB11 host cell functions and thereby increase the monoclonal antibody (mAb) titer up to 8.1 g/L/31 days under conventional 1-L bioreactor fed-batch conditions. Furthermore, the mAb produced by the DXB11/TAUT cells was comparable in quality …

Varied Productivity According To The Differences Between Targeted Locations Of Antibody Expression Vectors In Chinese Hamster Ovary Cells, Noriko Yamano, Jana Frank, Masayoshi Onitsuka, Kota Yoshitomi

Cell Culture Engineering XV

Chinese hamster ovary (CHO) cell lines are widely used in the pharmaceutical industry to produce therapeutic antibodies. However, trial and error cell selection methods are still used to construct high-producing cell lines. Exogenous genes are predicted to express differently depending on the expression vector integration sites. Chromosomal instability is one of the characteristics in CHO cells. We have previously constructed the CHO genomic bacterial artificial chromosome (BAC) library that is expected to cover entire the CHO-DG44 genome (Omasa et al., Biotechnol. Bioeng., 104, 986-994, 2009). The BAC-based physical map is a powerful tool to identify each chromosome and analyze …

The Renaissance Of Cancer Immunotherapy Is A Revolution For Patients, Ira Mellman

Cell Culture Engineering XV

Over just the past few years, cancer immunotherapy has transformed how we think about cancer care and cancer biology. This transformation is remarkable not only for its rapidity but also because cancer immunology itself had received little serious consideration for decades. At least three factors have been responsible: (i) the advent of actual data from the clinic as opposed to promising ideas from the laboratory; (ii) a more sophisticated understanding of the human immune response to cancer; and (iii) the realization that despite many years of effort targeting the oncogenic drivers of cancer, for the vast majority of patients the …

Implementation Of Raman Spectroscopy At Manufacturing Scale: Overcoming Modeling Challenges While Implementing Advanced Process Control, John Smelko, Thomas Matthews, Brandon Berry, Douglas Osborne

Cell Culture Engineering XV

In order to improve glycation control in our new high intensity platform process, we investigated implementation of Raman spectroscopy to monitor and control glucose at low levels throughout the process. Using Raman allows a way to measure glucose near real time which in turn allows for a more responsive and flexible control algorithm. Prior literature has shown connections between elevated glucose levels and glycation when in presence of high protein concentrations, which was indicative of our process. Using Raman along with advanced process control (APC), allowed us the flexibility to reduce glycation back towards historical process levels.

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