Upstream Processing

An Introduction to the FUTURA pico for Mini-bioreactors

Small bioreactors are gaining share in process development laboratories. It is vital that these bioreactors are realistic scale down models of larger systems and it is equally important that online sensors for measuring live biomass can also be used in these vessels. ABER, known and trusted for its sensors that measure live cell concentrations using capacitance technology and adopted in the world’s leading bio pharmaceutical companies from R&D through to cGMP processes brings you the new FUTURA PICO. Our Conventional…

Comparison of the Finesse SmartGlass™ and SmartVessel™ 3L Bioreactors: Engineering Data and Cell Cultivation Results

Finesse Solutions Inc. recently introduced the single-use benchtop scale SmartVessel 3L bioreactor, which was developed based on its re-usable counterpart, the SmartGlass bioreactor. With identical main geometrical parameters, the plastic bioreactor mimics the glass vessel with minor modifications for manufacturability. The following study focuses on the engineering data of both vessels, which have been determined with special focus on animal cell culture applications. Furthermore, data from material tests on leachables and extractables as well as results from cell cultivations using…

A Systematic Development Approach to Optimize and Control Biopharmaceutical Product Quality

The complexity of recombinant protein manufacturing processes, including expression systems, cell culture process conditions, and choice of media and feeds, present signifi-cant challenges to achieve desired product quality of bio-pharmaceuticals. To address these challenges, Fujifilm Diosynth Biotechnologies (FDB) has developed a unique systematic approach of combining “Media Toolbox” methodology, key chemical components, and cell culture bioprocess “know-how” to optimize and control manufacturing condi-tions that promote the desired product quality profiles of recombinant proteins. Case studies are presented to highlight the…

Targeting Desired N-Linked Glycosylation Profiles Through The Use Of Glycosylation Enhancing Feeds And High Throughput And High Resolution N-Glycan Analysis By Multi Capillary Electrophoresis

Glycosylation is a key product quality attribute for many biotherapeutic proteins expressed in CHO cells. N-linked glycans may display macro- and micro-heterogeneity; the degree of this variation can depend on several factors, including cell line, media/feeds, and process. As a consequence, it has often been challenging to achieve and maintain preferred glycosylation profiles from cell culture development through bioreactor scale-up. In order to address these challenges, we have developed a new feed technology in conjunction with a unique fed-batch process…

High Density Culture Strategies For Improved Scalability With Single-Use Systems

Improvements in single-use systems have allowed implementing high-density cultures in standard work flows. The current study shows integration of the Thermo Scientific™ HyPerforma™ Single-Use Bioreactor (S.U.B.) and the XCell™ ATF6 Single-Use (SU) System to achieve high-density cultures. Current results are compared against similar cultures using a stainless steel ATF6 system. The S.U.B. was able to support high-density cultures (>40E06 cells/mL) without modification to standard single-use components and maintained proper operating parameters. Scale-up criteria for both S.U.B. and ATF are provided…

CryoPod™ Carrier: Standardized Portable Cryogenic Handling of Cellular Therapies

Temperature control during sourcing, manufacturing, and delivery of cell therapies is crucial, as fluctuations can affect viability and function of the cells, and potentially, the overall efficacy and safety of the therapeutic product. In many cell therapy workflows, source cells arrive cryopreserved at LN2 temperatures (-150 to -196 °C) and post-manufacturing, cells are again cryopreserved and stored in vapor- or liquid phase LN2 prior to their use in the clinic. In either case, containers filled with dry ice (-50 to…

Efficient Optimization of CHO Cell Culture Medium and Feed for Increased Antibody Production

Traditional medium optimization strategies are labor intensive, costly, and time consuming. In this project, high-throughput screening technology was adopted along with statistical design to reduce costs and decrease time for optimizing culture conditions for maximized antibody production from a custom Chinese hamster ovary (CHO) cell clone. The presented work includes two phases: phase 1 for basal medium optimization and phase 2 for feed optimization. Phase 1 and phase 2 each have two rounds. A DoE screening methodology was used to…

An Intensified Perfusion One-Step Process with High-Density Cell Banks

Seed culture expansion is commonly performed in several consecutive batch cultures. Starting from a cryopreserved cell stock, the initial culture expansion is typically performed in shake flasks, while the final steps are performed, at greater scales, in bioreactor vessels. Such a procedure is time-consuming and labor intensive with multiple steps that introduce risk for mishandling and contamination. This work describes how the use of perfusion in the seed culture expansion process, in combination with the use of high-density cell banks,…

Development and Optimization of CHOgro® Transient Expression Technologies for High Titer Antibody Production in Suspension CHO Cells

During early stage drug development, quickly obtaining relevant candidate proteins through transient transfection can accelerate drug dis-covery. High titers are often obtained from Human Embryonic Kidney (HEK) 293 derived cell types; however, the use of different host cells between early stage transient and later stable protein production is a concern and can lead to the advancement of false-positive candidates. Chinese hamster ovary (CHO) cells are a desirable target cell type due to growth characteristics and a history of regulatory approval;…

Development of a Novel Cold Chain Tubing, FP-FLEX™, and Single- Use Freezing Bag for Working Cell Banks Enabling Closed-System Processing to Temperatures as Low as -196°C

Working cell banks (WCB’s) are commonly applied to initiate cell culture manufacturing campaigns for production of therapeutic proteins. These campaigns typically begin with inoculation of cells previously cryopreserved in vials. While vials are typically used to establish WCB’s and initiate manufacturing campaigns, they are not optimal for the growing demands of commercial production. Vials are small and filling/removal is performed through an open cap. This process leads to numerous manual operations and culture vessels, resulting in contamination risks and potential…