Regulatory agencies such as the FDA require the structure and amino acid sequence characterization of recombinant monoclonal antibodies (MAbs) to grant marketing approval. Characterizing such complex, inherently heterogeneous molecules is a significant analytical challenge that requires a broad array of physicochemical tests. Mass spectrometry (MS) is an essential tool for characterizing protein identity, functions, substrate specificity and amino acid sequence (AAS) of recombinant MAb biotherapeutics as it complements, or in some cases supersedes the utility of traditional biological methods. For some of the most important proteomic applications, the high sensitivity and accuracy provided by modern MS has allowed the unequivocal protein characterization…
Category: <span>Biologics Production</span>
The recent discovery of cancer stem cells in leukemia, brain cancer, and breast cancer has had a significant impact on cancer research and how cancers are thought to arise. Their discovery has resulted in unique, but pressing challenges that may ultimately only be resolved through the development of large-scale bioprocesses. The scarcity of these cells is currently impeding discovery of new cancer treatments that specifically aim to eradicate the population of tumour cells thought to be responsible for tumour growth and metastasis. Fundamental bioprocessing principles have been applied to develop scalable, large-scale cultures for cancer stem cells, to address this issue of cancer stem cell scarcity. Development of such bioprocesses differs significantly from other, more conventional cultures since it is the cells themselves that are of interest rather than the products of the cells (e.g., proteins)…
Controlled cell deposition by way of micrometer-sized jets are increasingly becoming a fiercely pursued area of research. We recently uncovered the ability to jet living cells using one such jetting methodology, now referred to as bio-electrosprays. This technique has never been explored for processing living cells until now. Electrosprays charge media within a needle, subsequently imposing an external electric field to assist in drawing the media into a micro-jet. The resulting instabilities assist in jet break-up, forming cell-bearing droplets. These droplets containing viable cells could be deposited onto a wide variety of biological and non-biological substrates. In this article, we report our developmental studies into this jet approach with a view to the successful handling and deposition of primary neonatal cardiac myocytes…
Proteins are widely used in research, medicine and industry, but its extraction from their natural sources can be difficult, tedious and expensive. Therefore, a simple and inexpensive system that allows large-scale production of safe recombinant proteins will always be highly desirable. Traditional production systems that use microbial, insect and mammalian cell cultures have drawbacks, in terms of cost, scalability and product safety. Several studies have shown that molecular farming in plants has many practical, economic, and safety advantages as compared to these conventional methods. Thus, the use of plants for recombinant protein synthesis is gaining wide acceptance…
A variety of affinity chromatography mechanisms exist that exploit either immobilized proteins or small molecule ligands. Among them, protein-A chromatography is widely used as a platform technology for a commercial-scale manufacturing of therapeutic monoclonal antibodies. Antigen, dye and specific ligand-columns are used to purify target proteins for research uses and diagnostic applications. It uses specific interactions between affinity columns and target proteins. In general, the samples containing a protein of interest, such as cell culture media, serum, and tissue or cell lysates, are loaded on to the affinity columns; the proteins will normally bind to the column due to high affinity…
The search for new and more effective drugs is a complex and challenging endeavor requiring a number of strategies for identifying and processing new drug candidates. A common tool used in this search is high-throughput screening (HTS) of a large library of small molecular weight compounds. In each of our HTS campaigns, more than 500,000 compounds are tested for biological activity against one of our many molecular disease targets in order to identify compounds of potential therapeutic interest. High-throughput screens can be divided into two major types: those using cells, and those not using cells. At least 50% of our screens are cell-based and require a consistent, uniform supply of cells throughout the screening campaign. The provision of the cells is a primary consideration in our cell-based assays and therefore, an automated cell culture laboratory was included in our HTS facility. This laboratory serves two functions: the preparation of cell banks, and the daily provision of microtiter plates containing cells for the HTS…
Enriched or pure oxygen is increasingly being used to improve mammalian and microbial cell culture productivity. Sterilizing filtration of the process gas stream is required to remove all microbial contaminants. The filters used to sterilize the gas stream must meet demanding requirements. They must be capable of withstanding sterilization and integrity testing after each use. Oxygen challenges filters because filter components may suffer corrosion and filter materials can potentially ignite under extreme conditions. So the components of filters used with pure oxygen should be tested and rated for pure oxygen service. This article will delve into the main issues involved in sterilizing enriched or pure oxygen gas streams…
Analytical tools for the characterization of protein identity and structure are fundamental to many fields of biochemical research. For the development of protein biopharmaceuticals, it is particularly important to measure modifications of the structure that may affect safety and efficacy. This application requires the analysis of large numbers of samples during process development. Small quantities of modifications must be detected in samples that are in the presence of more abundant native protein. Many kinds of analytical techniques are applied to this problem including peptide mapping, bioassays, liquid chromatography, spectroscopy, and so on. One of the most attractive tools is mass spectrometry, since essentially any change in the molecule is accompanied by a change in molecular weight. With the recent advent of readily accessible mass spectrometers capable of routine exact mass measurement, it is appropriate to consider the detailed requirements for this application…
The biologics market, although difficult to estimate, is currently thought to be in excess of $20 billion. In recent years, the growth in the novel therapeutics market has continued to exceed all but the most optimistic of expectations. The number of products in early stage trials may already be over 1,000, with an estimated 40 or so additional products in the process of finally being released to the market. The biologics market is led by relatively few “blockbuster” drugs, but the breadth of novel products continues to expand. This has resulted in exciting times for clinicians but has resulted in concern related to the bottleneck of production capacities for these drugs, as well as the pressure from healthcare agencies to reduce the cost of goods…
In today’s biopharmaceutical pipeline, monoclonal antibodies are a predominant modality for a broad range of clinical indications, including inflammatory disorders, oncology, and infectious diseases. More than two dozen antibody-based products are commercially available. In 2004, six of the 12 new biopharmaceutics that gained approval in the United States and Europe were antibody-based products. Most antibody therapies require high doses over a long period of time, which requires large amounts of purified product per patient. Therefore, manufacturing capacity to meet the demands of antibody production is a real challenge. It is desirable to have highly productive and consistent manufacturing processes. In addition, speed to market is critical to deliver health benefits to patients quickly and to achieve business success…