Expression vector and cell line engineering is the basis for expression and industrial production of biopharmaceuticals. The ultimate goal is to obtain clonal cell lines that secrete the protein of interest with high cell-specific productivity, and at consistently high levels over an extended number of cell generations, allowing for scale-up and cost-efficient large-scale manufacturing. Productivity and stability of expression are thus the prerequisites for developing commercially viable processes…
Category: <span>Manufacturing</span>
A basic engineering study has been performed to evaluate three different strategies for the production of monoclonal antibodies (MAbs) from Chinese hamster ovary (CHO) cells. Cells are expanded in suspension culture and are then inoculated into either fed batch or perfusion culture for MAb production. The first strategy, which is also the current industry standard, uses fed batch culture with the cells in suspension in a stirred tank fermenter. The second strategy uses perfusion culture with the cells immobilized on Cytopore™ microcarriers in a stirred tank fermenter. The third and final strategy is perfusion culture with Cytoline™ microcarriers in a fluidized bed fermenter. Perfusion cultures, while leading to a somewhat lower product titer, were characterized by a much smaller equipment footprint. This in turn led to a >30% reduction in investment costs and a 12% reduction in MAb production costs calculated over five years of depreciation and ten years of production time…
Building a MAb bioprocessing plant is a process which normally takes three years. Before starting the engineering work, a “locked” process is necessary. This means that all the steps have to be defined by volume, time, material balances and product yield. These calculations are based on the results obtained during process development. The titer and yield of functional, recoverable product determines the plant size. Optimal volumetric productivity [g/(liter reactor volume * day)] is of utmost importance. The main difference between fed batch and perfusion culture is that in the fed batch, a centrifuge is required for cell removal, whereas in perfusion culture, cell removal is performed by dead end filtration. This is possible because the majority of the cells are immobilised on the microcarriers, thus minimizing the burden on the clarification unit…
Membrane proteins such as hERG (human Ether-a-go-go Related Gene) and GPCRs (G-protein-coupled receptors) have been widely used as favorite targets for discovery of therapeutic drugs to treat cardiac arrhythmia, diabetes, epilepsy, cancer, glaucoma and many other indications. They are also widely used in cell-based assays to test new pharmaceuticals for safety in the early stages of drug discovery…
In order to move product development forward, the majority of biotech companies and academic institutions involved in cell-based therapies need new facilities in order to scale up production capabilities and comply with evolving regulatory requirements. Some institutions choose to use a contract manufacturing organization (CMO) to benefit from established expertise while others support their clinical development programs with their own dedicated production facility. The main challenges in establishing a dedicated pilot-scale production facility are described hereafter…
The biopharmaceutical industry has seen a major shift away from the use of serum and other animal-derived components in the manufacture of biopharmaceuticals. Guidance from the EMEA and FDA for the manufacture of biopharmaceuticals and medical devices encourages the use of “animal free” components to lessen concerns over contamination from adventitious agents such as prions, a cause of spongiform encephalopathy…
The development and commercialization of biopharmaceutical products is paving the way for significant growth in the pharmaceutical industry. Virtual and small biotech companies are researching and developing new biotechnology-based products that help large pharmaceutical companies backfill their pipelines. Many of these products have been in development for years and are finally reaching the point of commercialization. Because of the nature of many of these substances, they are sensitive to their environment, their packaging and their delivery systems…
The viable cell concentration is of prime importance in monitoring cell culture manufacturing processes. Of the available online biomass assays, the radio-frequency impedance (RFI) method has some clear advantages for manufacturing because it is an unambiguous reflection of viable cell biovolume rather than the total number of cells. This allows RFI to be used to control feeding rates or to maintain a constant level of biomass within the bioreactor. RFI is also suitable for measuring the live cell density in bioreactors when the cells are attached to microcarriers and to inert discs…
A patent permits the patent holder to keep others out. That is, a patent holder can prevent others from making, using, selling, offering to sell, or importing an invention within the United States and its territories and possessions. In the United States, these exclusionary rights generally last for 20 years from the date on which the patent application was filed. What the patent holder can keep others out of is defined by the “claims” of the patent. Patent claims describe the boundaries of the patent, much like a real property deed outlines the borders of a parcel of land. However, having a patent does not guarantee that the patent holder is free to use the claimed invention…