Genetic Engineering & Biotechnology News

AUG 2018

Genetic Engineering & Biotechnology News (GEN) is the world's most widely read biotech publication. It provides the R&D community with critical information on the tools, technologies, and trends that drive the biotech industry.

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22 | AUGUST 2018 | Genetic Engineering & Biotechnology News | GENengnews.com Joe Orlando, Ph.D. Development of an industry-scale bioproduc- tion process is a time-consuming, multistep operation that requires extensive biologi- cal and engineering expertise. Although the generation of unique recombinant cell lines hinders development of standard platforms, reduction of variables in cell lines, cell culture media, and growth vessels permits common strategies to achieve rapid and robust results. A holistic upstream solution can help ad- dress this challenge. In our turnkey and com- plete solution, we've reduced the number of variables by integrating multiple product lines, optimizing the process, and providing detailed protocols to guide users to a path of successful clone generation, process develop- ment, and scale-up while reducing resource requirements and timelines. Developing a robust bioprocess introduc- es many challenges for the user—the most significant being time and resources. Due to complex biological properties, development of a recombinant cell line requires working with a large number of cells and isolating a so-called "needle in a haystack." This process requires maintenance of nu- merous cell cultures a few times each week. After a cell line is isolated and selected, de- velopment of a robust bioreactor process re- quires multiple two-to-three-week-long runs at multiple scales—from small scale to pilot and production scale. These systems require daily sampling and extensive analysis to determine methods to improve the process for future runs. Two other challenges are process and product quality. Current process economics and efficacy needs dictate expected product titer and quality benchmarks at large produc- tion scale. Simply running the process does not guarantee that these attributes will be obtained. Alternatively, developing a basic platform into a system that can be utilized at scale for multiple molecules may take years. Recombinant Cell Line Development The CHOZN ® platform is a recombinant CHO (Chinese Hamster Ovary) mammalian cell expression system for the fast and easy se- lection and scale-up of clones producing high levels of recombinant proteins. It is supported by proven protocols, paired media and feed, as well as customized services for cell line de- velopment and cell line engineering. The host cell line is GMP-banked, and the history of the cell line is traceable and documented to support regulatory filings. The expression vector is IP-free, and the cell culture media is produced under strict GMP guidelines to support all regulatory filings. A description of the workflow and all the pro- tocols needed for successful clone generation is described in the CHOZN technical filing. Results from four independent cell line development projects are shown in Figures 1A & 1B. Cells were transfected with differ- ent dual-expression vectors containing light and heavy antibody genes. After selection, screening, and isolation, the final fed-batch titers for the best clones of each of the four projects are shown. Titers range from 2 g/L to over 4 g/L (Figure 1A). This range of titers is expected due to clone variability regarding molecule expressability, cell growth potential, and other biological factors. The corresponding specific produc- tivity results for these clones are shown in Figure 1B. Metrics such as these can aid in selection for specific applications (i.e., fed- batch or perfusion). All potential production clones should be evaluated for recombinant protein expres- sion stability in a three-month study. In the study shown in Figure 1C, the 10 highest expressing clones from a cell line develop- ment project were passaged twice a week for approximately three months, and different linages were evaluated for titer in fed-batch. The percentages of retained titer are shown in Figure 1C. Eight of the 10 clones CAYMAN CONTRACT SERVICES Your Vision. Our Expertise. Accelerated Discovery. DISCOVER MORE www.caymanchem.com/services contractresearch@caymanchem.com Bioanalysis & Assay Development Analytical Chemistry Medicinal Chemistry & Structure-Based Drug Design Chemical Synthesis MilliporeSigma Describes How to Carry Out Operations with Robust and Turnkey Platforms Demonstrating Process Scalability Bioprocessing Tutorial Joe S. Orlando, Ph.D., is principal scientist at MilliporeSigma. Website: www.emdmillipore.com Figure 2. EX-CELL® advanced cell culture feed optimization. Compared to the control, one of the regimens improved tiers 30%. Figure 1. Performance of CHOZN® GS clones. The CHOZN GS −/− cell line has been modified to eliminate endogenous glutamine synthetase (GS). Titers (A) and productivity results (B) from four different cell line development projects are shown. Percentages of retained titers (C) are shown for the 10 highest expressing clones from a cell line development project.

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