Downstream Processing

Genderless Sterile Connectors: How They Increase Flexibility, Reduce Complexity, and Drive Toward Standardization

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).

Three-Step Monoclonal Antibody Purification Processes Using Modern Chromatography Media

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…

Volume Reduction and Process Optimization with Cadence™ Inline Concentrator

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…

Adsorption Study of Egg-Derived Influenza Virus with Cellufine Sulfate Affinity Chromatography Media

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…

Purification of Monoclonal Antibodies from CHO Cell Culture Supernatants Using KANEKA KanCapA™

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…

New LEWA EcoPrime LPLC/BID – Analytical Accuracy at Pilot and Production Scale

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.…

Monoclonal Antibody Production – Viral Clearance and Host-Cell Protein Reduction Using a High-Capacity, Single-Use, Strong Anion-Exchange Device

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…

Scaling Up a MAb Purification Process Using Continuous Chromatography

In batch-mode chromatography, operating conditions do not allow full use of a media’s total capacity, leading to a subutilization of the media (1, 2). Sequential multicolumn chromatography (SMCC) is an open-loop process for the separation of multicomponent mixture buffers (3, 4). The concept behind SMCC is to perform different steps in smaller columns (from two to eight columns, compared with batch chromatography) connected in series, to maximize media use without product loss. BioSC® Lab is an automated system based on…

Single-Use Tangential-Flow Filtration Can Save Five Hours of Facility Time

Single-use technologies can be applied to tangential-flow filtration (TFF) applications within bioprocessing. In such applications, they can reduce the need for utilities, reduce capital expenditure during early phases of drug development, increase processing flexibility during process design and operation, and limit product cross-contamination risks. Read the full text of this application note in the PDF (Login required).

Modern Resins for a MAb Platform Process

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).