Bioreactor productivities are highly dependent on the process used to cultivate mammalian cells. These productivities directly affect the manufacturing plant capacity, and thereby the economics of production of monoclonal antibodies (MAbs). Historically, companies have chosen bioreactor process strategies that emphasize simplicity of scale-up at the expense of productivity, and conducted manufacturing using well-characterized and relatively straightforward batch processes. Such processes have successfully produced small or moderate quantities (ranging from ~100 g to ~ 1 kg per lot) of the desired antibody. Given the anticipated demand for large-scale quantities of MAbs (and the high stakes for the companies investing in these new biological entities), it is worthwhile to revisit these past selection strategies and see if — and under what conditions — they remain optimal today…
Category: <span>Biologics Production</span>
The revolution in biotechnology has led to 133 biotechnology-derived medicines being approved by 2001 with sales of $22 billion. This is less than 10 percent of today’s total pharmaceutical market, but it is a rapidly growing sector. Biologics are predicted to grow to nearly $50 billion by 2008. Marketed biopharmaceuticals include several blockbuster products with multibillion-dollar sales. In recent years, biotechnology-derived therapies represented 10 percent to 20 percent of all new approved molecular entities and hundreds more are in development, including nearly 200 proteins in late-stage trials. Microbial and mammalian expression systems are typically used to produce biotherapeutic proteins (many companies are also working on transgenic expression systems). Microbial cultures (typically, Escherichia coli or yeast) are used to produce smaller, less-complex proteins or those where specific modifications, especially glycosolation, are not required…
Recombinant monoclonal antibodies (rMAbs) are the predominant biotherapeutic protein under development today. FDA requires the structure characterization if rMAbs and other recombinant proteins to grant marketing approval. Characterizing such complex, inherently heterogeneous molecules is a significant analytical challenge that requires a broad array of physico-chemical tests. This article reports the use of reversed phase high-performance liquid chromatography (RP-HPLC) with on-line electrospray ionization mass spectrometry (ESI-MS) to rapidly determine the glycoform composition and the heavy chain C-terminal lysine heterogeneity of an intact rMAb. In addition, a novel multidimensional chromatographic platform was developed to investigate the two-dimensional, size exclusion chromatography (HPSEC) separation of the rMAb followed by RP-HPLC (HPSEC-RP-HPLC) with on-line ESI-MS analysis. Such analyses can characterize, identify, and confirm the structure of an intact rMAb…
Long-term growth of the biopharmaceutical industry is increasingly relying on outsourcing to overcome the current capacity constraints, especially for monoclonal antibody production. Companies are often reluctant to commit to building multimillion dollar manufacturing facilities for potential products with no guarantee of approval. Therefore to offset risks, companies will enter into contract manufacturing arrangements…
Tissue engineering is an emerging area of biotechnology that will provide replacement tissues for patients, as well as complex, functional biological systems for research and testing in the pharmaceutical industry. A new research area of tissue engineering is the investigation of how living cells interact with and respond to synthetic biomaterial surfaces. The clinical developments that underlie that research include a number of novel tissue-engineered medical products (TEMPs)…
Monoclonal antibodies constitute a significant percentage of the protein-based therapeutic molecules currently in clinical trials. The broad applicability and proven commercial success for this class of molecules suggest a larger future market potential. The current biopharmaceutical manufacturing capacity is widely anticipated to be a rate-limiting factor in the growth of the biotech sector. Because antibody therapeutics represent such a large part of this market, and because the therapeutic dosages of antibodies tend to be greater than most biopharmaceuticals, there is an immediate need for novel antibody manufacturing approaches that deliver significantly greater productivity…
Over 25 years have elapsed since Kohler and Milstein electrified the immunology community with their article describing the reliable preparation of monoclonal antibodies (MAbs) by fusing immune splencytes with immortalized myeloma cells. This discovery not only garnered the pair of scientists a Nobel Prize, but also led to the development of a technology which has yielded a number of important therapeutic, prophylactic, and diagnostic products for in vivo human use, and hundreds of in vitro diagnostic products. Some of these products proved to be significant in meeting previously unmet medical needs, and a few have been commercial successes. But the path, from Kohler and Milstein’s discovery to commercial products, was discontinuous and a bit bumpy, and the technology continues to evolve…
Organizations developing biopharmaceuticals are often faced with the challenge of developing, as rapidly as possible, a production system for a recombinant protein or antibody intended for use in clinical trials. For expression of antibodies and other proteins with complex post-translational modifications, Chinese hamster ovary (CHO) cells are often the host of choice. However, isolation of CHO cell lines producing even moderate levels of a protein of interest is usually a lengthy process due to the need for at least one and usually several gene amplification steps. Gene amplification, which is usually accomplished through the dihydrofolate reductase (dhfr)/methotrexate system, is a requirement for most CHO expression vectors because the absolute expression level from each copy of an integrated expression plasmid is generally very low…
Large scale genomics spurred the development of massively parallel methods of automated DNA purification and sequencing. These methods started with the 1962 development of a 96-well microtiter plate for miniature-scale serology studies. This simple laboratory device has since been greatly modified and extended to include numerous specialty multiwell plates contructed and/or coated with different materials for various purposes. The original 96-well (8×12 matrix) has expanded to include 384-well and higher densities. More recently, functionality and versatility have been greatly augmented by the incorporation of a filter, or thin membrane, into the bottom of the well. These multiwell microfilter plates can thus be employed in a flow-through mode, in addition to the familiar “put in” and “take out” pipetting and rinsing steps associated with traditional enzyme-linked immunosorbent assay (ELISA) microtiter plate methods…