Biologics Production – Page 29 – BioProcessing Journal

Application of Chemically-Stable Immunoglobulin-Selective Sorbents: Capture and Purification of Antibodies with Resolution of Aggregate

A growing number of separations’ scientists and process developers are looking beyond protein A sorbents for capture and initial purification of monoclonal antibodies. A variety of strategic and operational goals have prompted examination of alternative immunoglobulin-selective sorbents. Most broadly, many workers wish to eliminate design considerations associated with leached protein A. Also cited is a preference for sorbents that can withstand stringent cleanup using 1 M sodium hydroxide. In some applications, it is desirable to avoid the low-pH elution conditions typically employed with protein A sorbents — conditions that can foster aggregate formation. In still other cases, the target antibody may bind poorly to protein A. Finally, there may be interest in evaluation of immunoglobulin-selective sorbents less costly than protein A sorbents…

Biologics Production

chromatography immunoglobulin-selective sorbents monoclonal antibody optimization process development purification separation

Bioprocessing Aspects of Neural Stem Cell Production in Bioreactors

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

bioreactor cell culture media neural stem cells scale-up stem cell production stem cells

Quality Control, Preparation, and Protein Stability Issues for Blood Serum and Plasma Used In Biomarker Discovery and Proteomic Profiling Assays

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

automation biomarkers high-throughput mass spectrometry profiling proteomics quality control stability

Scalability of a Disposable Bioreactor from 25 L-500 L Run in Perfusion Mode with a CHO-Based Cell Line: A Tech Review

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

bioreactor contaminant control continuous perfusion disposable bioreactor optimization scalability single-use

Cell Transfection Using Calcium Phosphate Ceramic

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

calcium phosphate ceramics ceramic transfection gene expression gene-activated matrices recombinant gene transfection

Biological Packaging for the Global Cell and Tissue Therapy Markets

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

biological packaging cell stress cryopreservation hypothermic preservation preservation solution storage

Single Protocol for the Selective Isotopic and Chemical Labeling of Protein Kinases in Insect Cells for NMR and Crystallographic Studies

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

expression isotopic labeling labeling nuclear magnetic resonance spectroscopy optimization phosphorylation protein kinase X-ray crystallography

A Flow Cytometric Assay for Rapid, Accurate Determination of Baculovirus Titers

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

BEVS flow cytometry high-throughput optimization protein expression recombinant proteins viral titer

Integration of Centrifuges with Depth Filtration for Optimized Cell Culture Fluid Clarification Processes

One of the biggest challenges in the production of recombinant therapeutic proteins, monoclonal antibodies, and vaccines is the clarification and separation of the product (typically a protein) from the cell culture or fermentation broth. The desired product is present in low concentrations and must be efficiently separated from the other components present in the bioreactor fluid. An overall objective in developing a clarification process is to achieve the highest level of product recovery (yield) and contaminant removal with the fewest number of unit processes. Understanding how each operational step affects the performance of the next step downstream is the challenge at hand. Centrifugation, in combination with depth filtration, is gaining acceptance as the preferred method for the removal of cells, cell debris, colloids, insoluble precipitants, aggregates, and other materials found in mammalian cell culture and bacterial fermentation fluids…

Biologics Production

centrifugation clarification contaminant control depth filtration process development separation

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

apoptosis bioreactor cell cycle cell stress cellular engineering mammalian cell culture productivity

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