Downstream

Removal of Endotoxins – From Bench to Process Scale

Endotoxins or lipopolysaccharides (LPS) are highly toxic components of the cell wall of Gram-negative bacteria, which are often present in significant amounts in bacterial cell expression systems such as E.coli. A number of methods have been adopted for the removal of endotoxin based on adsorption, in particular ion exchange chromatography. Although downstream processing can significantly reduce endotoxin levels in the product, efficient and cost effective removal of residual endotoxin from biopharmaceutical preparations remains a challenge. Prometic Bioseparations have developed an…

We Meet Your Single Use Process Monitoring Requirements

Innovation is at the core of all pharmaceutical, biopharm and bioprocess applications. With extensive experience in the biopharmaceutical industry — and particular expertise in single-use bio-pharmaceutical equipment — PendoTECH delivers a line of pressure sensors, control systems and software for measuring, monitoring and data collection in bioprocess applications and other areas where the products provide a unique process solution. Committed to meeting the specialized needs of life-science laboratories and bio-pharmaceutical manufacturers, PendoTECH technical consultants review your company’s applications in the…

Single-Use Automation: Unlocking the True Potential of Single-Use Technology

When single-use technology first arrived on the scene in the late nineties, the processes that it was applied to were relatively simple and low risk. These included simple filtration and storage applications of buffers and cleaning solutions. With these filtration steps typically involving lower value, non-blocking solutions of easy to dissolve salts, automation would have been seen as an unnecessary over- complication. It was thought sufficient to have an operator watch the process to make sure nothing became disconnected, a…

Purification of Antibody Fragments and Single Domain Antibodies With Amsphere™ A3 Protein A Resin

The standard capture purification of full-size, classical antibodies is typically performed with Protein A affinity chromatography. Binding of the Protein A affinity ligand to the “Fc” region of antibodies (Abs) (abbreviations are defined at end of paper) takes place at the juncture of the constant domains 2 and 3 of the Ab heavy chains (Lewis et al. 2008). This high affinity binding primarily involves hydrophobic interactions. Antibodies that belong to the same subclass have greater than 95% homologous Fc-regions, allowing…

Purification of Fab and scFv Using High Binding Capacity TOYOPEARL® AF-rProtein L-650F Resin

Protein L based affinity chromatography is used for the capture of antibodies and antibody fragments that do not bind to protein A. Unlike protein A and G, which bind to the Fc region of immunoglobulins (IgGs), protein L binds through interactions with the variable region of an antibody’s kappa light chain. TOYOPEARL AF-rProtein L-650F is an affinity chromatography resin that combines a rigid polymer matrix with a recombinant protein L ligand, which is derived from the B4 domain of native…

Continuous Capture Chromatography, A simple, Robust Approach to Move From Batch to Continuous Capture

Increasing demands for the production of biopharmaceuticals, such as monoclonal antibodies (mAbs), vaccines and recombinant proteins, require new, innovative purification approaches with higher efficiency and productivity while maintaining or reducing costs. One of these approaches is continuous capture chromatography. Continuous capture chromatography, also known as CaptureSMB was developed by ChromaCon AG. CaptureSMB is a 2-column periodic counter current chromatography (2-PCC) process where the first column is fully loaded and breaks through to the second column. The first column is eluted…

Quantum ReNu SU Technology® Validation Guide and Performance Testing Summary

As the use of disposable systems within biopharmaceutical manufacturing has moved from niche preclinical applications to large scale commercial processes. This has led to the requirement for novel technologies that improve processing throughput and allow manufacturers to increase their output. To this end, WMFTG has developed a new pumping technology to enable end-users to meet their downstream processing needs for unit operations such as chromatography and tangential flow filtration (TFF). Quantum allows end-users to use novel peristaltic technology for their…

Achieving Continuous Downstream Bioprocessing Through Multi-Column Chromatography (MCC)

In recent years, improving downstream manufacturing has become an emerging challenge to the biopharmaceutical industry. Initially, the bottleneck in biopharmaceutical manufacturing existed in the output of bioreactors. Due to advances in cell line development and upstream process (i.e., perfusion), the output of bioreactors has increased at a much faster pace than downstream processing capacity. Therapeutic biologics, including monoclonal antibodies (mAbs), represent the fastest growing segment of the pharmaceutical market. By 2019, the biological market is expected to exceed 20% of…

Optimizing Unit Operations In Biopharmaceutical Manufacturing

There are many critical material-handling challenges regarding flow rates, pressure levels and the prevention of shear in the chromatography, virus filtration and TFF processes that are used in the manufacture of biopharmaceuticals. In order for these unique operations to be implemented successfully while handling fluids that can be sensitive, delicate and expensive, the operator must be aware of their specific operating characteristics and choose a pumping technology that can meet the strict demands for successful operation. While lobe and peristaltic…

Pegasus™ Prime: Robust Retention after Pressure Interruptions (Stop and Start)

Virus filtration is a vital part of the viral clearance strategy in bioprocesses. In filtration, size exclusion mechanisms are orthogonal to other inactivation or removal techniques and target the physical dimensions of the virus to achieve a high degree of virus reduction. Any flow or pressure interruption during filtration has been shown to increase the risk of virus passage. This application note reviews the PegasusTM Prime virus removal filters and demonstrates robust, high viral clearance in the presence of prolonged…