August 2015

From the outside, you’d be forgiven for thinking of a bioprocess container as a mere box. Yet it is this box — or magic box — that transports valuable fluids, buffers, and media along your in-process storage and handling supply chain. Read the full text of this application note in the PDF (Login required).

This technical note presents the benefits of using a single-use heat exchanger and its areas of application. Successful cell culture on a large scale requires close control of the culture environment in terms of temperature, pH, removal of waste products, and addition of fresh nutrients. To produce high yields of a desired protein, precise control over such parameters at every step in a process is required. Additionally, FDA regulations require that various steps be taken to control bioburden and maintain sterility. Read the full text of this application note in the PDF (Login required).

From drug and biologic development services to delivery technologies to supply solutions, we are the catalyst for your success. With over 80 years of experience, we have the deepest expertise, broadest offerings, and the most innovative technologies to help you get more molecules to market faster, enhance product performance, and provide superior, reliable manufacturing and improved results. Catalent is your strategic partner for biologic drug development success. Read the full text of this application note in the PDF (Login required).

Cell banks must be extensively characterized to assess the cell source with regard to its identity, the presence of other cell lines, and adventitious/ endogenous agents or molecular contaminants. However, the tests used in this process are species specific and can vary depending on the history of the cell line and the type of media components that may be present. The following are assays that need to be considered. Read the full text of this application note in the PDF (Login required).

Working cell banks (WCBs) are commonly applied to initiate cell culture manufacturing campaigns to produce therapeutic proteins. Those campaigns typically begin with the inoculation of cells previously cryopreserved in vials. Although vials are typically used to establish WCBs and initiate manufacturing campaigns, they are not optimal for the growing demands of commercial production.

We demonstrate the success of large clinical-scale culture of human adipose-derived mesenchymal stem cells in an industrial single-use vessel at a 3.75-L working volume. The vessel provides a precisely regulated environment for the growth of stem cells under carefully controlled simulated physiological conditions. The monitoring of the cultures allowed us to accurately regulate their performance, optimizing their growth while at the same time demonstrating appropriate stem cell markers coupled with differentiation assays performed at the end of the culture period. With a view to future cellular therapy exploitation, every step from initiation to conclusion in the expanded bioreactor vessel was performed with single-use disposables. Read the full text of this application note in the PDF (Login required).

As the largest international network of harmonized good manufacturing practice (GMP) product testing laboratories, Eurofins BioPharma Product Testing delivers a true local service experience. Eurofins BioPharma Product Testing offers the most complete range of testing services, harmonized quality systems, and laboratory information management systems (LIMS) to more than 2,000 pharmaceutical and biopharmaceutical companies worldwide. Read the full text of this application note in the PDF (Login required).

This application note describes the cultivation of Chinese hamster ovary (CHO) suspension cells in the Finesse SmartGlass vessel bioreactor with a maximum working volume of 2.0 L. Using chemically defined minimal media, cell densities of up to 7.44 × 10 6 cells/mL were achieved. Recombinant secreted embryonic alkaline phosphatase (SEAP) expression was induced by medium exchange and temperature shift. Maximum SEAP activities of up to 63 U/mL were reached. A novel stirred glass bioreactor suitable for cell culture applications at benchtop scale was designed by Finesse Solutions, Inc. With a maximum working volume of 2.0 L, this bioreactor is controlled by the Finesse G3Lab controller. Read the full text of this application note in the PDF (Login required).

Continuous improvements in growth media and cell line viability have resulted in increased biomass concentrations in biopharmaceutical production processes, making the downstream purification step more challenging. Body feed filtration (BFF) has proven to be a successful method of solving similar issues in other industries (Figure 1). This robust technology is now available as a harvesting solution for biotechnology applications ( 1 ). Read the full text of this application note in the PDF (Login required).

Historically, single-use connections involved a male and a female half. These were two distinctly different parts that mated with each other. Genderless connections — those in which the connector halves are identical in design — bring multiple benefits to modular single-use systems. Read the full text of this application note in the PDF (Login required).

This application note describes a monoclonal antibody (MAb) purification process using the expanded MAb purification toolbox of GE Healthcare’s Life Sciences business, covering modern chromatography media (resins) for standard and challenging purification tasks. The most established approach for purifying MAbs is a three-step process, where the initial capture step using a protein A medium is followed by two polishing steps using cation- exchange (CIEX) and anion-exchange (AIEX) media (Figure 1a). However, for more challenging MAb purifications, an expanded MAb purification toolbox is beneficial. Here, a multimodal AIEX medium was used as an alternative in final polishing of a MAb with low monomer stability (Figure 1b). Using the described approach, the set criteria for the polishing steps were achieved, with a recovery of >90% over each step and an aggregate content <1% in the final product. Read the full text of this application note in the PDF (Login required).

Pall’s patented single-pass tangential-flow filtration (TFF) technology simplifies current TFF processes ( 1 ). It eliminates the conventional TFF recirculation, allowing an efficient single-pass operation without mixing or foaming issues and exposes the product to low shear, which optimizes the processing of fragile and shear-sensitive molecules. The single-pass feed flow rates are lower than those of conventional TFF, minimizing the hold-up volume and maximizing product recovery. It allows for higher final concentrations because processing limitations due to minimum working volumes are optimized ( 2 – 4 ). Read the full text of this application note in the PDF (Login required).

An efficient process for purifying virus particles is important when developing a vaccine. Cellufine Sulfate affinity chromatography medium has been used for manufacturing viral vaccines such as influenza virus, rabies virus, and Japanese encephalitis virus. Here we describe how to purify egg-derived influenza with Cellufine Sulfate media. Figure 1 shows a typical chromatogram of inactivated influenza virus A strain (H7N7) from allantoic fluid with Cellufine Sulfate media. Table 1 shows that adsorbed virus particles are eluted from the medium easily with high purity. Read the full text of this application note in the PDF (Login required).

Developing robust and scalable purification processes for monoclonal antibodies (mAbs) can save time and money. KANEKA’s protein A chromatography resin, KANEKA KanCapA™, offers the required high-performance characteristics to develop platform purification processes for mAbs: high binding capacity, high alkaline stability, good elution profiles, low nonspecific binding, and robustness in column packing. Here we provide purification data of five different mAbs (two humanized IgG1s, two humanized IgG2s, and a chimeric IgG1) using KANEKA KanCapA™. The Chinese hamster ovary (CHO) cell culture supernatants were kindly provided to us by our customer. Read the full text of this application note in the PDF (Login required).

The new LEWA EcoPrime® LPLC chromatography platform combines proprietary fluid design and LEWA Intellidrive® technology to deliver unrivaled batch-to-batch reproducibility and accuracy. The new EcoPrime system features our open-architecture automation software and is built on a quality control of the industry’s most vertical supply chain. A single LEWA system has the range of up to three conventional LPLC units while providing higher accuracy. That gives users great flexibility, more floor space, and plant-friendly integration with simple software customization and updating. Read the full text of this application note in the PDF (Login required).

Strong anion-exchange (Q) chromatography is an industry standard for polishing purification steps of monoclonal antibody (MAb) production. It is a proven technology that removes viruses and host-cell proteins (HCP) from process feed streams in flow-through mode. NatriFlo HD-Q device adsorbers feature a hydrogel membrane, which combines high dynamic binding capacity and salt tolerance to deliver best-in-class performance. This makes it possible to rapidly process-large sample volumes using a small disposable device. This article examines the ability of NatriFlo HD-Q adsorbers to clear virus and HCP from partially purified (post–protein A) MAb solutions. Read the full text of this application note in the PDF (Login required).

Purolite is using new technologies to produce homogeneous beaded agarose for protein purification at large scale. Particle sizes range from 40 to 90 μm. The resins cover applications from capture to polishing and demonstrate improved pressure–flow properties (Figure 1). The porosity structure is optimized for medium-sized proteins such as monoclonal antibodies (MAbs). Read the full text of this application note in the PDF (Login required).

Specifically engineered to meet the requirements of large-scale bioreactors, OPUS® 45-cm and 60-cm columns match the performance of traditional self-packed columns while delivering the cost and labor savings expected from ready-to-use prepacked columns. Read the full text of this application note in the PDF (Login required).

POROS™ XQ is a next-generation strong anion- exchange resin with a fully quaternized amine surface chemistry. It was designed to retain all the beneficial performance attributes of POROS HQ resin, such as resolution capability, flow properties, and physical stability, with the added benefits of higher capacity and salt tolerance. Those were realized by optimizing the pore structure, which drives better access of biomolecules to the surface charge, and by optimizing the amount of surface charge (the ligand density) as measured by ionic capacity. Better access to a higher amount of ionic capacity increases dynamic binding capacity (DBC) and improves salt tolerance, because the resin can withstand higher amounts of counterion and still allow effective protein binding. Read the full text of this application note in the PDF (Login required).

Design of Experiments (DoE), also called experimental design, is a statistical approach to process development that has gained wide acceptance in the biopharmaceutical industry. DoE is used to reduce development costs by speeding up the design process and to optimize the parameters of a particular step in the manufacturing process. With its inherent ability to take into account multiple variables at once, a designed experiment is also used to minimize process variation leading to a more robust manufacturing process. Read the full text of this application note in the PDF (Login required).

A polishing step for the purification of a monoclonal antibody (MAb) using TOYOPEARL NH2-750F was developed. In general, anion-exchange resins can be used in bind and elute (B/E) mode as well as in flow- through (FT) mode. Both options were evaluated. To increase the amount of aggregates of the test sample, a MAb was aggregated by acidic incubation and subsequently diluted to 1 g/L in loading buffer. Read the full text of this application note in the PDF (Login required).

In July 2015, Avid Bioservices debuted its new state-of-the-art biomanufacturing facility in Tustin, CA. Designed to be one of the first US late phase 3 and commercial production facilities for the exclusive operation of single- use equipment to produce biologics, the 40,000-ft 2 build-out of an industrial 110,000-ft 2 warehouse provides the perfect setting to install state-of-the-art modular cleanroom bioproduction suites. This new facility incorporates the relatively recent concept of converting an existing building in to a new bioproduction facility, known as a “brownfield” design. The end result is an innovative and flexible design that will support the transition of your program from late phase 3 into commercial production. Read the full text of this application note in the PDF (Login required).

Advances in cryopreservation techniques have been crucial for the substantial progress achieved in many fields, including cell biology research, drug discovery, bio-banking, and assisted reproduction. While cryopreservation techniques have markedly improved, downstream cell thawing techniques have thus far been neglected, even though proper thawing of cryopreserved materials is essential for optimal cell viability. The success of high-promise fields such as cellular therapy and regenerative medicine require reproducible and standardized handling of the therapeutic cells, which includes thawing both during manufacturing and prior to patient administration, to ensure effective patient response. BioCision recognized cell thawing as a process that has been historically overlooked but is critical to the success of the overall process. Consequently, the company developed the ThawSTAR™ Automated Cell Thawing System – an instrument that senses the contents of each cryogenic vial and customizes a rapid thaw of the...

Boehringer Ingelheim BioXcellence™ is your dedicated biopharmaceutical contract manufacturer. As a leading biopharmaceutical contract manufacturer with more than 35 years of experience, it has brought more than 20 biopharmaceutical products to market. Boehringer Ingelheim BioXcellence™ offers tailor-made contract development and manufacturing services to the biopharmaceutical industry, providing the entire production technology chain from DNA to fill and finish under one roof at its facilities in Biberach (Germany), Vienna (Austria), Fremont (USA), and Shanghai (China). Boehringer Ingelheim BioXcellence™ can secure product supply throughout the entire product life cycle transferring customer projects at any stage, delivering to almost any scale. Boehringer Ingelheim BioXcellence™ makes outsourcing easy. Read the full text of this application note in the PDF (Login required).