Expression & Production

Maximizing Protein Expression in CHO Suspension Cells Through Transient Transfection

Transient transfection in mammalian cell lines provides an avenue for researchers to bridge the development bottleneck and shorten the time to usable protein. The method also maintains post translational modifications crucial for biotherapeutic function. Chinese Hamster Ovary (CHO) suspension cells are especially suited for high yield production of recombinant proteins, despite being refractory to commonly used transfection methods (e.g. 25kDa linear PEI). Mirus Bio has developed a more effective alternative, the TransIT-PRO® Transfection Kit. Transfection efficiencies are affected by many…

Novel Plastic Bag for Monoclonal Antibody Final Bulk Storage to Actively Prevent Oxidation

Essentially, human protein such as monoclonal antibody is quite susceptible against oxidation. Nevertheless using the oxygen barrier containers, oxygen within the protein solution and oxygen leaking through the closure port of container affect to the protein. To minimize oxidation of monoclonal antibody, novel plastic bag for monoclonal antibody final bulk storage has been developed. The concept “Active” means that removing oxygen not only passively from the circumstances through the barrier layer within the container (or bag), but also actively removing…

Bio-Simplex™ Single-Use, Aseptic PreCulture Systems

The growth and aseptic transfer of a microbial pre-culture from seed vial to bioreactor is a critical process step in the production of BioPharmaceuticals. The pre-culture step is performed in a biological safety cabinet or laminar flow hood, which adds time and significant cost and to the process. Saint-Gobain Bio-Simplex™ Single-Use Pre-culture Systems provide a sterile environment for cell culture and aseptic transfer of all fluids in and out of each cell culture vessel.

Characterization of a Biomanufacturing Fermentation Process Using a New DOE Approach: Definitive Screening Designs

Traditionally, a bioprocess is characterized using a two-step approach involving factor screening designs followed by response surface designs. This approach is very costly and time consuming since it requires a significant number of runs. Recently, Jones and Nachtscheim (2011) described a more efficient design that allows for factor screening, resolution of two-way interactions as well as estimation of quadratic effects in a single-step. The design, termed Definitive Screening has not yet been validated on a bioprocess. Here, traditional and Definitive…

Improving Cell Culture Productivity with a SEGFLOW® Automated On-line Sampling and Feed System

A SEGFLOW® automated on-line sampling and feed system (SEGFLOW) was evaluated for its impact on Chinese Hamster Ovary (CHO) cell culture process performance in bench-scale stirred tank bioreactors. The SEGFLOW was integrated with a YSI® Biochemistry Analyzer and a nutrient feed pump allowing real-time glucose analysis and automated glucose feeding to be performed for the test batch. In contrast, a batch using daily manual sampling and bolus feed protocols, was performed as a control. For the SEGFLOW culture, media glucose…

Efficient Production of Recombinant Antibody Derivatives in Pseudomonas flourescens

P. fluorescens serves as an attractive expression host for the production of stable and biologically active antibody fragments. A proprietary strain of P. fluorescens has been specifi cally developed as a protein production platform to enable rapid identification of strains capable of expressing high titers of soluble, active protein, and refl ects the additional sophistication required of engineered antibody derivatives being developed today. Pfēnex Expression TechnologyTM employs a toolbox of defined P. fluorescens strains containing custom-designed combinations of genetic elements…

ATF Perfusion-based Production Platforms for Biopharmaceuticals at Good Grip – Concept s for Robust Control and Operation

Continuous Perfusion is comparatively unpopular rather than considered as a first choice production platform for complex biopharmaceutical glycoproteins. Major concerns that have been addressed are mainly related to risk of process failure during long-term operation, complexity and scalability of cell retention devices, supply and definition of consecutive lots and, finally, handling of continuous harvests in downstream processing. We have cleared-up these stereotypes and designed a robust modular perfusion-based production platform for biopharmaceuticals demanding highest quality at industrial scale. Main components…

Towards the Use of Statistical Modeling to Predict Cell Culture Performance of Hydrolysates

Hydrolysates are complex media supplements composed of many as well as different types of components. Predicting cell culture performance of a particular lot remains an elusive target, other than by small scale cell culture testing which consumes valuable time, effort and resources. In order to reduce the need for incoming acceptance tests by our customers, FrieslandCampina Domo plans to develop a simple analytical assay that accurately predicts cell culture performance. Our Quality by Design project has provided information at the…

Effect of process condition on the Mycoplasma rentention of cell culture media filtersMethods

A Mycoplasma contamination event can have a major impact on a biopharmaceutical manufacturer. The loss of a cell culture due to a contamination incurs significant costs that can be attributed to both the initial bioreactor set-up and to the subsequent decontamination. Production facility throughput may be affected and in the worst cases the ability of the manufacturer to supply patients with medicines. Mycoplasma are extremely small in size and lack a cell wall giving the cells some flexibility that enables…

Engineering Analysis of Mixing in ATMI’s Bioreactors Using Computational Fluid Dynamics

The Integrity® PadReactor™ and Nucleo™ systems are a single-use bioreactors specifically designed to fulfill the needs of cell culturists. They are perfectly suited to laboratory environments, process develop¬ment centers, clinical material supply and flexible GMP manufacturing. The bioreactor vessel, which offers comparable functionality to classical stirred tank bioreactors, is a single-use bag integrating an internal paddle mixing and sparger system. This innovative bag design allows a non-invasive connection to the system. The paddle is enclosed in a medical grade ULDPE…