The contamination of microbiological media by mycoplasmas such as Acholeplasma laidlawii is not a recent phenomenon. It has been a major problem with animal-derived sera since the 1980s and has been a concern in the management of cell cultures for decades. The main culprit of serum contamination was the inadequate blood collection methodology and was eliminated with the introduction of hollow collection needles. In addition to the introduction of an improved collection method, serum was filtered with 0.1…
Analytical
Robustness of Parvovirus–Retentive Membranes and Implications for Virus Clearance Validation Requirements
Generic validation is conceivable only through a thorough understanding of the parameters affecting the performance of a process step. In this paper, we provide a detailed example demonstrating the robustness of a virus filtration step. As a first step towards the establishment of a generic validation package for a monoclonal antibody, the robustness of clearance of PP7 across the ViroSart CPV filter was evaluated by changing several critical operational parameters using a simple one-off experimental design. Two different…
Which Impeller Is Right for Your Cell Line?
When growing microbes or animal cells in a stirred-tank reactor, it is critical to choose the impeller type that is best suited to your process. Select the wrong impeller, and you could make chop suey of your filamentous fungi. Pick the right impeller, and you could greatly increase yields of your fussy mammalian cultures such as Chinese hamster ovary (CHO) and Vero kidney epithelial cells. With a wide range of impeller designs to choose from, how do you…
Many Considerations in Selecting Bioproduction Culture Media
Not long ago, the ability to support efficient large-scale culture of cells was the main factor in choice and development of production media. However, a number of new performance demands have been imposed on production media (as listed in the “Key Factors†box). These new criteria arise from such sources as the demand for increased efficiency in a number of production operations, goals invoked by new quality initiatives, and a more science-based approach to process development. Not only…
Monitoring ATP Status in the Metabolism of Production Cell Lines
Development of industrial cell culture processes for production of recombinant proteins seeks high efficiency, reproducibility, and predictability. Usually the time allowed for process development is short, during which culture conditions and scale-up protocols must be defined so as to maximize cell productivity and yield while minimizing process scope and overall costs (1). Although scientific literature describes various methods that increase productivity of a cell culture by reducing and arresting cell growth or weakening cell physiology (2), the cells must be…
Considering Cell Culture Automation in Upstream Bioprocess Development
Cell culture automation has become more common in drug discovery and research applications, thereby freeing bench scientists from repetitive work as well as improving the consistency of their cell culture processes (1). Mammalian cell culture is used in the production of biopharmaceuticals, where developing a production cell line requires a large amount of repetitive manual work for bench scientists. With the increasing importance of biologics in today’s pharmaceutical market, throughput and efficiency are crucial in developing a production cell line.…
Why Do So Many Biopharmaceuticals Fail?
Biopharmaceutials and the processes used to make them are exceedingly complex, and the path to developing new therapeutics is a high-risk endeavor. The emphasis today is on controlling product quality, safety, and efficacy through understanding biological mechanisms, key product attributes, and process parameters. Such information is also crucial for guiding development efforts to improve chances of success in the clinic and for gaining regulatory approval. Analytical methods provide the foundation for acquiring such knowledge. Efforts devoted to developing meaningful and…
Maximizing Data Collection and Analysis During Preformulation of Biotherapeutic Proteins
Preformulation research, a critical component in the development of biotherapeutics, explores the effects of variables such as pH, ionic strength, and excipients on the solution behavior of a protein. This activity can greatly assist in guiding downstream formulation development, and it provides valuable information concerning protein stability, solubility, and structure. Successful preformulation research leads to identification of potential protein degradation pathways and development of robust formulations with acceptable product shelf-lives. PRODUCT FOCUS: PROTEINSPROCESS FOCUS: DOWNSTREAM PROCESSING, FORMULATIONWHO SHOULD READ: FORMULATIONS,…
Novel Excipients Prevent Aggregation in Manufacturing and Formulation of Protein and Peptide Therapeutics
Protein and peptide therapeutics may undergo numerous physical and chemical changes during manufacturing, shipping, storage, and administration that can adversely alter drug potency and safety. Earlier concerns focused upon denaturation (unfolding), oxidation, and deamidation of certain key aminoacyl residues. Recently, aggregation has emerged as a key issue underlying multiple deleterious effects for peptide- or protein-based therapeutics, including loss of efficacy, altered pharmacokinetics, reduced stability and product shelf life, and induction of unwanted immunogenicity. As a result, the US FDA and…
How to Improve Your Implementation of Two-Dimensional Preparative HPLC
The biologics and natural product industries rely heavily on separation technology. Sample analyses are undertaken on the analytical scale, and isolation and purification are undertaken at the preparative scale. Key target components are often isolated to provide standard reference materials for future product quality assurance testing. These products are often very complex mixtures, requiring separation systems to have a high peak capacity for both analytical and preparative scale separations. A technique gaining popularity among companies that require the isolation of…