mammalian cell culture – Page 2 – BioProcessing Journal

Mouse Hybridoma Cell Culture in a Protein-Free Medium Using a Bio-Mimicking Fish-Tail Disc Stirred Bioreactor

Because the Lambda MINIFOR bioreactor provides good mixing of cell culture, nutrients, and gases without any damaging hydrodynamic forces by using a bio-mimicking “fish-tail“ disc stirrer, it can be successfully applied for the cultivation of bacteria and yeast, insect, plant, and mammalian cells. However, reports on its application in mouse hybridoma cell culture using protein-free media is non-existent in the scientific literature. Therefore, this study describes preliminary findings of the Lambda MINIFOR bioreactor suitability in mouse hybridoma cell culture and antibody production using the SP2/O-Ag14-CB.Hep-1 mouse hybridoma cell and the PFHM-II protein-free medium as models. Results verified 2.45 × 10⁶ viable cells/mL as the highest cell concentration, 86% as maximum cell viability, 0.0156/h as the exponential growth rate, 44 h as cell population doubling time, a stable phenotype measured by limiting dilution after 2.5 months, no antibiotic and antifoam requirements, 71.4% of IgG SDS-PAGE purity in the cell culture harvested supernatant, 38.68 ± 22.29 µg/mL, 39.23 ± 10.66 pg/cell/day, up to 99.5% of purity (sample measured by SDS-PAGE and SE-HPLC) after an IgG capture step based on protein A-Sepharose, a low pH incubation, and size-exclusion chromatography, no molecule aggregation, specificity for the CKTCTT epitope (located in the HBsAg “a” determinant), an IgG affinity constant equal to 1.11 × 10¹⁰ M^-1, and < 78 pg mouse DNA/mg of IgG. In conclusion, this study corroborated a cumulative CB.Hep-1 mAb production of 1.77 g/15 days and validated the usefulness of the Lambda MINIFOR bioreactor in mouse hybridoma cell culture in protein-free media for research applications...

Biologics Production

fish-tail disc stirrer hybridoma mammalian cell culture protein-free media stirred-tank bioreactor

Monitoring Viable Cell Density in Bioreactors Using Near-Infrared Spectroscopy

Viable cell density (VCD), the quantitative assessment of living cells, is commonly determined by laborious and inaccurate off-line cell counting methods. Single-function in situ probes have been developed using various technologies including optical density, radio frequency and dielectric permittivity. Optical density measurements predict total biomass but are sensitive to cell debris accumulation and inherent media turbidity. Near-infrared (NIR) has the advantage of being able to measure many key analytes in the cell culture simultaneously. NIR has been used to acquire real-time measurements of glucose, glutamine, glutamate, and media nutrients such as amino acids and metabolites (e.g., lactate and ammonia). NIR probes can be sterilized in place, and real-time measurements can be acquired throughout the mammalian cell culture processes…

Biologics Production

bioreactor mammalian cell culture near-infrared spectroscopy process monitoring viable cell density

Profiling Intracellular Monoclonal Antibody Expression for Improved Cell Culture Characterization and Process Optimization

Many laboratories have utilized cell-free systems or prokaryotic systems designed to produce biological molecules with single polypeptide chains, limited folding requirements, and without glycosylation. The yeast systems are used to generate glycoproteins; however, their glycosylation profiles are vastly different from those of mammalian cells. Without significant glycoengineering, the yeast-produced recombinant glycoproteins may be unsuitable as therapeutic molecules. As such, the use of mammalian cells is still the preferred method to produce complex biological molecules…

Biologics Production

antibody expression characterization flow cytometry mammalian cell culture process development specific productivity

Testing Strategies for Mammalian Cell Banks

Contamination by adventitious agents (bacteria, fungi, mycoplasma, and viruses) represents potential safety risks for biologics produced in mammalian cells. Bacterial and fungal contaminations are usually easy to detect in culture medium due to changes in pH and visual indicators such as color and opacity. Mycoplasma contamination has been detected in 15–35% of cell lines deposited in some cell culture collection. This is because mycoplasma contaminations often cause little changes that can be readily detected by visual inspection. However, bacterial, fungal, and mycoplasma contamination can be more effectively controlled than viral contamination by careful screening of initial parental cell banks, proper environmental monitoring, along with ongoing testing…

Cell & Tissue Banking

contaminant control mammalian cell culture raw materials qualification safety testing viral clearance viral contamination

Filtration of Enriched and Pure Oxygen Key to Increasing Cell Culture Application Yield

Enriched or pure oxygen is increasingly being used to improve mammalian and microbial cell culture productivity. Sterilizing filtration of the process gas stream is required to remove all microbial contaminants. The filters used to sterilize the gas stream must meet demanding requirements. They must be capable of withstanding sterilization and integrity testing after each use. Oxygen challenges filters because filter components may suffer corrosion and filter materials can potentially ignite under extreme conditions. So the components of filters used with pure oxygen should be tested and rated for pure oxygen service. This article will delve into the main issues involved in sterilizing enriched or pure oxygen gas streams…

Biologics Production Manufacturing

aerobic fermentation integrity testing liquid challenge test mammalian cell culture microbial cell culture oxygen transfer sterile filtration

Current Therapeutic Antibody Production and Process Optimization

In today’s biopharmaceutical pipeline, monoclonal antibodies are a predominant modality for a broad range of clinical indications, including inflammatory disorders, oncology, and infectious diseases. More than two dozen antibody-based products are commercially available. In 2004, six of the 12 new biopharmaceutics that gained approval in the United States and Europe were antibody-based products. Most antibody therapies require high doses over a long period of time, which requires large amounts of purified product per patient. Therefore, manufacturing capacity to meet the demands of antibody production is a real challenge. It is desirable to have highly productive and consistent manufacturing processes. In addition, speed to market is critical to deliver health benefits to patients quickly and to achieve business success…

Biologics Production Manufacturing

antibody purification antibody therapeutics chromatography mammalian cell culture process development process optimization

Conquering Design Challenges in the Construction of Immunomedics’ Multiproduct Cell Culture Bioreactor Facility

In the last few years, we have seen many biotech products approved by FDA. These products have gained public awareness because of their ability to treat several debilitating diseases with very minimal side effects, and thereby impact the quality of life for many people. As a result, the biotech industry is constantly in the news for its successes and programs to develop new therapeutics for many unmet medical needs. Immunomedics, Inc., a New Jersey biotechnology company, recently completed an expansion project that included new bioreactor manufacturing suites and support laboratories. Building on the company’s existing headquarters site and fully integrating the new capacity into the existing operational facility, the project spanned two years and was completed in 2003…

Manufacturing

bioreactor facility design mammalian cell culture process development project management

Characterization and Release of Raw Materials Used in Upstream Processes for Production of Monoclonal Antibodies by Mammalian Cell Culture

Since the mid-1970’s, when Kohler and Milstein first discovered the process by which myeloma cells and splenocytes could be fused to produce monoclonal antibodies (MAbs), a whole new world of important therapeutic, prophylactic and diagnostic products has opened up, bringing in huge benefits for patients and manufacturers. The total sales of therapeutic MAbs reached more than $13 billion in 2005. Sixteen of the 18 FDA-approved MAbs came to the market after 1997, and over 150 are currently in clinical development, suggesting their increasing medical importance and the remarkable, recent advancements in development technology…

Biologics Production

characterization growth promotion test mammalian cell culture monoclonal antibody raw materials raw materials qualification

Innovation Through Biomanufacturing Platform Development

Total market share of biopharmaceuticals is estimated to increase from $33 billion now to more than $45 billion in 2007. These numbers are accounted for by the 64 products approved by European and US regulators and some of the 500 products currently under clinical evaluation. More than 2,000 products are in discovery and preclinical development. Monoclonal antibodies (MAbs) and recombinant glycoproteins constitute a major part of these new biotech leads. The estimated demands for MAbs alone are more than 6,000 kg per year in 2006. Currently, 16 MAbs are licensed by the U.S. Food and Drug Administration (FDA) for pharmaceutical use and more than 130 are in clinical trials. This fast-growing class of biotherapeutics is expected to reach worldwide sales of more than $15 billion per year in 2008. In the coming years, mammalian cell culture technology will remain the production system of choice for MAbs and other recombinant glycoproteins. Therefore, efficient, cost-effective production systems need to be in place to meet the demands…

Biologics Production Manufacturing

contract manufacturing mammalian cell culture monoclonal antibody optimization process development recombinant glycoproteins scalability

Cell Engineering Blocks Cell Stress and Improves Biotherapeutic Production

Within the biopharmaceutical industry, mammalian cell culture is extensively used to manufacture a various biopharmaceutics uncluding antibodies, interferons, hormones, crythropoietin, clotting factors, immunoadhesins, and vaccines. The market for monoclonal antibodies (MAbs) alone is expected to grow 30% a year and reach sales of nearly $6.5 billion in 2004. The vast majority of these biotherapeutics are secreted glycoproteins obtained from mammalian cell lines such as: Chinese hamster ovary (CHO), human embryonic kidney 293 (HEK-293 or 293). NS0, and baby hamster kidney (BHK). As is the goal with most commercial products, biotechnologists strive to generate these valuable proteins in the highest yields possible in order to utilize mammalian bioreactor facilities efficiently…

Biologics Production

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