Over the past decade there has been a steady increase in the number of biotherapeutics requiring high doses and long term administration. Most notable among these are monoclonal antibodies (MAbs) and fusion proteins comprised partially of antibody molecules. Column chromatography is a commonly applied purification method for downstream processing of biotherapeutics, and there is considerable pressure to process much greater volumes at a faster rate. For recombinant proteins and MAbs, a variety of chromatographic methods are employed, including affinity, ion exchange, hydrophobic interaction, and to a lesser extent, immobilized metal affinity and gel filtration. Improving process control for chromatography operations is essential for biopharmaceutical manufacturers to process larger volumes and overcome capacity shortfalls. As the past ten years have seen increasing volumes of MAb-based drugs, there have been significant innovations to address growing productivity requirements. Dominant among these has been high throughput media capable of isolating product at faster rates than previously achievable…
Category: <span>Biologics Production</span>
Monoclonal antibodies (MAbs) destined for use in drug-specific assays must meet specific binding criteria, and therefore require much more in the way of development than many protein-specific antibodies. This article describes one way to facilitate the development of well-characterized, high specificity MAbs. Essentially the same techniques have also been used for producing MAbs that are cancer markers and MAbs that are specific for infectious agents. Monoclonal antibody development, as in other endeavors, requires clearly defined goals and an examination of proposed methods to attain them. This has been succinctly stated by the phrase “Start with the end in mind.” Unfortunately, the vast majority of hybridoma development could be more appropriately described by Yogi Berra, “If you don’t know where you are going, you will wind up somewhere else.” In most laboratories, fusions are carried out with the goal of reducing the amount of cell culture and handling required and using the minimum amount of screening. Little or no consideration is given to ensuring that single clones are subjected to the screening tests. Under these conditions, it is little wonder that dozens of fusions can be carried out, each with the same result — no specific antibody found…
lation components that stabilize the molecule in order to provide the desired product storage stability. Generally, an aqueous formulation is preferred; however, the instability of proteins, both physical (e.g. aggregation) and chemical (e.g. deamidation and oxidation), often necessitates the development of lyophilized formulations. In these formulations, selection of the appropriate stabilizing cryoprotectants, lyoprotectants, and bulking agents is critical. Accelerated stability studies are typically used to evaluate the effect of a single factor at a time in order to identify the optimum pH, buffer, and stabilizing excipients. This approach is limited in that many independent time-consuming experiments must be run, the results are obtained only at the evaluated set points, and additional experiments are required to assess potential interactions between the evaluated factors…
A clinical-scale manufacturing process has been developed for the ex vivo expansion of autologous cytolytic T lymphocytes (CTLs) directed against cells infected with the hepatitis B virus (HBV). The process is based on the Rapid Expansion Method (REM) technology originally developed at the Fred Hutchinson Cancer Research Center in Seattle, WA by Greenberg and Riddell. Preparations are underway to initiate a company-sponsored Phase I clinical trial in which REM will be used to expand rare autologous HBV-specific CTLs that will then be infused to patients chronically infected with HBV. Earlier studies have shown that such patients mount only a weak CTL response to HBV. Chronic hepatitis B can lead to severe liver damage such as cirrhosis and hepatocellular carcinoma. By infusing clinical-scale quantities of autologous HBV-specific CTLs into chronic HBV patients, it may be possible to boost the immune system so that it can control the viral infection…
Foreign substances, such as disease-causing viruses and other infectious agents are recognized by the body’s immune system as invaders. Parts of the spectrum of our defenses are antibodies that bind these antigens and direct their destruction by other elements of the immune system. Antibodies have two very useful characteristics. First, they are extremely specific; that is, each antibody binds to one particular antigen. Second, the body retains the memory of encounters with antigens, enabling it to reproduce an antibody response to a later attack by that that same agent or disease. These properties make it possible to develop vaccines. It is the first trait of antibodies, their specificity, which makes monoclonal antibody (MAb) technology so valuable. Not only can antibodies be used therapeutically, to protect against disease, they also can help diagnose a wide variety of illnesses and can detect the presence of infectious agents (including biological threat agent). The use of MAbs specific for biological warfare agents in fielded assays requires maintaining a high-quality and economical supply of these reagents…
With the advent of whole cell-based therapeutics has come a growing standardized quality control and quality assurance of the processes employed for developing and manufacturing cellular materials, similar to the controls over traditional drugs and biologicals. Cellular therapeutics present unique process and quality control challenges due to the innate complexities of living cells, making it important to use whole cell assays to provide detailed pictures of the status and consistency of cell preparations that will be used to treat patients. This article illustrates how a cellular assay from Guava Technologies addresses these issues…
Cell culture was first devised at the beginning of the 20th century as a method for studying the behavior of animal cells free of systemic variations that might arise in the animal both during normal homeostasis and under the stress of an experiment. During the past thirty, thousands of academic and for-profit organizations have come to rely on cultivation of animal cells as the basic foundation to perform biomedical research and large-scale biomanufacturing. Their success is directly dependent upon the reproducible production of high quality cell culture products. The complexity of the mammalian cell, its growth and storage requirements, and the need to maintain pure and uncontaminated cultures is a constant challenge to those involved with in vitro cell culture…
Contract manufacturing of recombinant protein drugs and vaccines, as well as other biopharmaceuticals, has been the focus of considerable interest during the past decade. Fueled by a strong clinical development pipeline, primary manufacturing of biopharmaceuticals on a contract basis has attracted multinational industrial concerns willing to invest on the promise of potentially higher returns than are experienced in the production of traditional small molecule drugs. Biopharmaceutical contract manufacturers have made significant contributions to the development and subsequent commercialization of a few highly successful products. However, despite strong growth, consistent profitability has been elusive. The market has changed overr the past decade as customer projects progressed from process development through market launch. Now that several preeminent market players have successfully made the difficult transition from clinical to commercial supplier, what has been learned and how is the market expected to evolve over the next five years?…