Expression Platforms

Protein Scaffolds

The recent success of monoclonal antibodies (MAbs) as therapeutic agents to treat cancer, multiple sclerosis, rheumatoid arthritis, and other chronic inflammatory and autoimmune disorders (Table 1) has catapulted these once difficult-to-develop molecules to the forefront of modern molecular medicine (1, 2). The size of the global MAb market in 2008 was valued at almost US$28 billion. Industry analysts predict that the size of the MAb market will grow to almost $68 billion by 2015, with the largest growth occurring in…

A High-Yielding, CHO-K1–Based Transient Transfection System

Biotherapeutics have emerged as effective treatments for many diseases. It’s estimated that every year hundreds of new biotherapeutic candidates enter development (1). Stable transfection of host cells to establish high-producing cell lines is the approved method for generating clinical-grade recombinant biologics. However, biotechnology companies needing to speed up their developmental timelines are increasingly relying on material generated using transient transfection (1, 2). Unlike stable gene expression (SGE) that requires several months of laboratory and process work, transient gene expression (TGE)…

Screening Yeastolate Raw Material Used in Insect Cell Culture Media

Insect cell culture is widely used as an industrial platform in recombinant protein production for research and diagnostics (1, 3,4,5). Many commercial yeastolate products are intended as nutritional supplements for not only insect cell culture, but also for mammalian cell culture and bacterial fermentation applications (6,7,8,9,10,11,12,13,14,15). Yeastolate plays an important role in modern biological production because it can be very effective in promoting insect cell growth and enhancing production of recombinant proteins. The product is known to contain amino acids,…

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…

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…

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…

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.

A Host Cell Protein Assay for Biologics Expressed in Plants

    Host cell proteins (HCPs) can be present in significant amounts in biological products through copurification with a recombinant protein-drug substance. Purified active ingredients developed for human use must be free of all contaminants — including residual HCPs — to minimize the incidence of immunogenicity against a drug product or its trace contaminants (1).   HCP assays allow for monitoring host-related impurities during product development and process development. Most are in the enzyme-linked immunosorbent assay (ELISA) format for detecting…

How an Obscure Asian Rodent Took Over Biotechnology

The earliest domestic rodents were cavies (“Guinea pigs”) kept as food animals in the Andes since 5000 BC (as shown by mummified cavies) and carried to Europe in the 16th century (Elizabeth I had one as a pet). Although rats weren’t domesticated until the mid-19th century (a byproduct of the blood sport of rat-baiting), mouse selection and breeding began many centuries earlier. Selection of unusually colored mice was first documented ~1100 BC in China, and breeding such “fancy” mice as…

A Decade of Animal Cell Culture

Eukaryotic cells are fragile and finicky, requiring very specific culture conditions and nutrients to survive, grow, and be productive in an ex vivo environment. Even so, they have become vital to the biopharmaceutical industry’s ability to make complex biological products — overtaking yeast as a production system around 1990 and surpassing bacteria in the number of associated product approvals five years later (1). Since then, they have become even more useful, expanding their reach into the vaccine world. Mammalian cell…