Figure 1. At recent scientific conferences on downstream processing, there was much discussion about the need to reduce process volumes. Many manufacturing sites are space-limited and to take advantage of chromatographic process improvements, they must be made without the need to increase buffer and holding tankage. One significant opportunity to increase process throughput is to replace conventional with higher capacity ion exchange resins. Recently developed high capacity resins allow more protein to be loaded without the need to replace existing…

Biopharmaceutical companies continue to wrestle with how to approach the implementation of process analytical technology (PAT). The accelerated adoption of many new technologies has benefited from the identification of a “killer app,” an early application that enables a readily achieved breakthrough in capability. This can be a totally new capability (e.g., e-mail on the internet) or a major step-change (e.g., 10× or more) in an existing capability. Flashes of inspiration cannot be achieved at will, but we can identify elements…

While industry is rapidly shifting from stainless steel vessels and hard-plumbed processes to single-use bioreactors and TFF filters, the transition between the multiple process steps remains complicated and time-consuming, often requiring fluid transfers, disrupting sterile barriers, and cleaning protocols. The simple solution to a complex process is connectivity: a continuous flow-path that interconnects all the process steps by means of a sterile singleuse process manifold. Spectrum Laboratories incorporates process filters, reservoir bags, flexible tubing, and other critical elements into a…

Upfront Chromatography A/S is a Danish company with significant expertise in the development of customized industrial processes. Successful operation of industrial separations provides proven scalability for biopharmaceutical manufacturers. Upfront Chromatography’s Rhobust™ technology has been developed to maximize the effectiveness of capture adsorption steps, using second-generation expanded-bed technology. Rhobust™ consists of Mixed-mode ligand chemistry (robust adsorbents with added selectivity) Tungsten carbide densified agarose that enables robust operation of expanded bed Dedicated hardware designs (robust fluid distribution systems without meshes, frits, or…

Cell counting and viability assessment are routine but essential parts of bioprocessing procedures, and accuracy, reproducibility, and standardization are essential to successful outcomes. Traditional methods produce variable results because they rely on subjective discrimination of dye-stained cells using a hemacytometer and microscope. Substantial operator-to-operator variability in the cell counts is common, even when one researcher examines the same sample twice. Guava technologies offers a solution: a simple, accurate, and reproducible absolute cell counting and viability assay that overcomes many of…

The development of single-use bioreactor technologies is focused on bringing new designs onto the market for effective oxygen transfer and cultivation of not only cell culture processes, but also microbial cultures. The advantages of disposable technologies are: ease of use, prevention of cross-contamination, labor and capital investment savings, reduced qualification and validation, and increased flexibility. The costs of disposables are lower compared with conventional stainless-steel bioreactors because the capital investment is lower. Therefore, the need is reduced for qualification and…

Figure 1. How well does absorbance correlate with cell density and viability measurements in batch and fed-batch cell culture? As expected in science and nature, the answer depends on a set of factors. Many people working with batch or fed-batch cell culture will reference discrete offline measures of cell activity. Total cell density (TCD), viable cell density (VCD), and percent viability can be evaluated offline using a hemocytometer and accepted trypan blue exclusion method. Cedex® and Vi-CELL™ are two examples…

Figure 1. CellPrime rTransferrin recombinant human transferrin provides an animal-free alternative to iron salts for industrial cell culture. It is a recombinant analogue of human transferrin expressed in Saccharomyces cerevisiae. Supplied as a human holo-transferrin analogue, CellPrime rTransferrin binds specifically to the transferrin receptor, thereby facilitating iron uptake into cells for optimal cell culture performance. Here we describe cell culture data demonstrating that CellPrime rTransferrin shows equivalence to human transferrin (hTf) and superiority to bovine transferrin (bTf) in stimulating cell…

Photo 1. Demand for single-use bioprocess systems in the biotech and pharmaceutical industry has increased significantly in recent years. Among other reasons — such as operations, sourcing, and validation — their lower initial investment capital makes disposable systems very attractive for the pharmaceutical industry. When Wave Biotech introduced the first wave-action reactor/mixer, it was seen by some as the start of single-use technology. Once containers become single-use, all other parts of the manufacturing process ideally have to follow the same…

Figure 1. This study describes a simple procedure for improving insect cell yields in a benchtop cell culture bioreactor. Here, yields of Spodoptera frugiperda (Sf-9) cells were increased by nearly 29% through monitoring dissolved carbon dioxide (dCO2) levels in the culture and adding air to the vessel headspace to reduce dCO2. The method can also be used to maximize yields in a wide range of mammalian cell types. Introduction Producing high yields of protein from insect cells usually requires maintaining…