Engineering Commons^™

Use Of Quality By Design Principles For Development Of Upstream Process Control Strategy, Canghai Lu, Girish Pendse

Cell Culture Engineering XV

A systematic approach was developed using the QbD (Quality by Design) principles to study and characterize monoclonal antibody upstream production processes. This approach comprises of risk assessment of upstream process parameters, small scale model development and qualification, process characterization as well as the determination of CPPs (Critical Process Parameters) and PARs (Proven Acceptable Ranges). A high throughout analytical method to measure N-linked glycans and a Chemometric method to calibrate the results were developed in order to overcome the analytical bottleneck. These studies improved the process understanding, explored the multivariate interactions, and established the relationship between CPPs and CQAs (Critical Quality …

Small-Scale Comparison Of Pseudoperfusion Feeding Strategies Using Basal And Concentrated Feed Media, Iona Bettinardi, Leda Castiho

Cell Culture Engineering XV

Perfusion has long been the industrial choice for the production of unstable proteins, and nowadays is being intensively studied also for stable proteins due to its ability to keep high viable cell densities and high volumetric productivities over long operation times. However, conventional perfusion is fed with basal medium at high dilution (or perfusion) rates, causing a dilution of the protein of interest (POI) and generating large volumes of harvest to be processed in the purification steps.

In this work, small-scale pseudoperfusion experiments were performed to compare the conventional perfusion strategy using basal culture medium (TC-LECC, Xell AG) with a …

Challenges In The Use Of Scale-Down Models For Understanding And Mitigating Process Variations Of A Monoclonal Antibody Production Process, Peter Russo

Cell Culture Engineering XV

Scale-down models are commonly used to execute process characterization studies, as well as to screen raw materials and to conduct investigations in support of manufacturing operations. During the clinical manufacture of a monoclonal antibody, consistent product quality was maintained; however, large variations in bioreactor harvest titer (> 2X) were observed between lots. Root cause analysis of this variation did not establish a linkage between culture performance and manufacturing execution or deviations. Further, small scale satellite reactor performance displayed lesser variability; thereby eliminating seed train and raw materials as potential root causes. Small scale investigations, using multiple scale-down models, were not …

Omics Approach For Generating A High-Yield Cho Cell Line Producing Monoclonal Antibodies, Wei Chi, Hsuan-Pu Chen, Dalton Chen, Hsin-Lin Lu, Bor-Shiun Chen, Chao Yi Teng, Chien-I Lin, Hsueh-Lin Lu, Chi-Chen Hsu, Sheng Jie Huang

Cell Culture Engineering XV

Chinese hamster ovary (CHO) cells are extensively used for the industrial manufacture of therapeutic antibodies. Generating high producing cell lines for secretory protein production requires knowing the bottleneck in the cellular machinery for protein expression. Integration site of gene of interest (GOI) is one of the important factors that influence the protein productivity. Even though screening of cells randomly integrated GOI can select high producing cells, the selected cell might not stable due to the chromosome instability. Here, we would like to look for host integration sites where GOI is high yield and stable by screening a single copy integration …

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 …