BioProcessing Journal Posts

Neurodegenerative diseases such as Parkinson’s disease and multiple sclerosis, along with injuries such as stroke affect millions of individuals worldwide and costs healthcare systems billions of dollars each year in North America alone. The diseases result from the death of specific cell types within the central nervous system. Current treatment efforts have focused primarily on alleviating symptoms using pharmaceuticals. However, recent advances in our understanding of these conditions, coupled with advances in biology, genomics, transplantation, and biochemical engineering are making cell therapy (the transplantation of viable cells to replace dead cells) more attractive as a potential avenue of treatment…

Biologics Production Cell & Gene Therapy

There is an increasing emphasis in clinical and translational research on the discovery and development of biomarkers that are indicative of a disease state. While biomarkers are not exclusively proteins, the emergence of new mass spectrometry platforms combined with the human genome databases has rejuvenated the search for biomarker proteins, especially in readily available body fluids such as blood. There is currently a tremendous need for an improved ability to “mine” the full depth of the proteome in a high throughput manner. To advance clinical proteomics, methodologies are needed that can accommodate higher throughput while facilitating the ability to observe large numbers of protein events…

Biologics Production Research

Single-use, disposable components offer many advantages in the manufacturing of biologics. They are clean and ready to use when supplied, which obviates the need for sterilization and decreases the requirement for services such as water for irrigation (WFI) systems and steam generators. Disposable components are not used for subsequent operations, eliminating the chance of cross contamination between process runs. Long lead times for equipment installation can be avoided because the need for stainless steel equipment is reduced or eliminated. Systems are less complex, therefore engineering requirements are also reduced. There is no need for clean-in-place (CIP) or steam-in-place (SIP) operations, along with the associated piping, valves, controls, or pressure rating of vessels. Moreover, the use of disposable components reduces the complexity of validation…

Biologics Production Manufacturing

The non-viral introduction of genes into mammalian cells (transfection) is of growing interest for tissue engineering and as an alternative to the use of viral transfer of recombinant genes. The introduction of a foreign gene into cells in vivo is often limited to the use of viral vectors such as adeno or retroviruses. Viral vector may present several disadvantages or side effects that can be disastrous, and the selection of cells that are transduced by the virus is very poor. A number of non-viral vectors have been explored and used to date: lipid-based carriers, hydrogel polymers, polycationic lipids, polylysine, polyornithine, histones, and other chromosomal proteins, such as hydrogen polymers and precipitated calcium phosphate. Most of these vectors are usable in vitro but are difficult to apply in vivo, especially when local transfection to a specific cell line must be obtained…

Biologics Production

Current in vivo gene therapy (GT) approaches are beginning to demonstrate significant clinical and safety limitations that may ultimately reduce their therapeutic utility. In particular, the potential for systemic toxicity due to the antigenicity of the gene transfer vector, the prospect of insertional mutagenesis/oncogenesis during gene transfer, and the possibility of germ line transfer of the transgene are issues raising concern. One promising alternative to gene therapy that mitigates these clinical and safety issues is gene-based cell therapy (GBCT), in which autologous cells are removed from a patient and modified ex vivo for a desired characteristic prior to reimplantation. By transferring the transgene ex vivo, many of the issues surrounding the in vivo use of the transfer vectors are reduced and issues surrounding germ line transfer can be practically eliminated…

Cell & Gene Therapy

Singapore’s vision is to become a global hub for the biomedical sciences (BMS) with world-class capabilities ranging from basic and clinical research to manufacturing and healthcare delivery. This vision encompasses pharmaceuticals, biotechnology, medical technology, and healthcare services. Singapore’s BMS initiative was launched in June 2000 with the goal of developing the industry into a key pillar of Singapore’s economy. It is overseen by a Ministerial Committee chaired by Deputy Prime Minister Dr. Tony Tan and implemented by an executive committee led by Mr. Philip Yeo, who is chairman of the Agency for Science, Technology, and Research (A*STAR) and co-chairman of Singapore’s Economic Development Board (EDB)…

Manufacturing Research

The last decade witnessed remarkable scientific and technological advances in a number of scientific disciplines, including cell biology, microbiology, molecular biology, oncology, virology, infectious diseases, diagnostic technologies, analytical chemistry, instrumentation, and informatics. These advances have had a major impact on medicine, which has experienced fantastic progress in improving disease diagnosis, treatment, and overall patient care. Despite the advances in developing ever more sophisticated technologies and increasing the understanding of disease, new maladies continue to emerge. This is especially true for infectious ailments. Despite great developments in epidemiology, diagnostics, and agent detection technologies, as well as a comprehensive understanding of the biology of many known infectious agents and their virulence factors, we also are witnessing a dramatic increase in the number of new agents and diseases….

Research

The globalization and sustained growth of the biotechnology market has brought the issue of biological packaging to the fore, particularly for those companies invested in cell and tissue bioproducts, such as engineered tissues and cells used for cell therapy. Biological packaging can be defined as the sum total of the physical device, temperature regulating and monitoring systems, type of preservation solution, and storage protocol(s) necessary to maintain cells or tissues in a “state of suspended animation” during transport or storage. The ideal biological package provides for the transport of cells and tissues throughout the global marketplace while maintaining both the viability and the function of the biological system at levels equivalent to those measured prior to shipment. Cells and tissues are currently shipped and stored under hypothermic (4–8ºC) or cryopreserved (–80 to –196ºC) conditions. These two processes have remained relatively unchanged over the past several decades, limiting their utility in the storage of modern bioproducts. However, recent evolutions in biological packaging have begun to provide scientific and financial benefits to researchers, clinicians, and corporate entities…

Biologics Production Cell & Gene Therapy

The human protein kinase superfamily is one of the largest and most important families of enzymes. More than 500 distinct kinases, classified in about 20 families on the basis of their primary structure similarity, have been identified to date. Protein kinases regulate a variety of biochemical pathways in cells through phosphotransfer reactions, playing pivotal roles in most signaling and regulatory processes, such as gene expression, proliferation, cell motility, and angiogenesis. Deregulation and/or mutational modification of protein kinase activity, leading to aberrant protein phosphorylation, is implicated in a variety of diseases, particularly cancer, making protein kinases important drug targets. A number of specific protein kinase inhibitors has been developed recently and more than 30 compounds are currently in clinical development or on the market. Many of these inhibitors are small-molecule compounds that compete with ATP for the highly conserved ATP binding site of the kinases. The development of highly selective and potent ATP-competitive inhibitors is driven by structure-activity relationship (SAR) studies, with X-ray crystallography and nuclear magnetic resonance (NMR) spectroscopy playing an important role in the understanding of the mechanism of inhibitor binding to the active or inactive forms of protein kinases…

Biologics Production

Recombinant protein expression using the Baculovirus Expression Vector System (BEVS) is a powerful tool for the production of therapeutics, diagnostics, and reagents. To maximize efficiency of protein production, and thereby reduce costs, it is important to optimize the production parameters. A crucial step in optimization is determining the best multiplicity of infection (MOI) for the system in use. Factors that can affect the MOI include the recombinant baculovirus itself as well as cell line type and media composition. Typically the titer of a viral stock is determined in a standard manner, and then that titer is applied to each and every parameter tested; for instance, titering the virus on a Spodoptera cell line in a serum-containing media, and then using those data to determine the MOI used to infect Trichoplusia cells in a serum-free media formulation. The results may suggest that either the Trichoplusia cell line or the media formulation is inadequate for protein expression when, in fact, the MOI was incorrect for that particular combination…

Baculovirus Expression Technology Biologics Production