The implementation of single-use technologies for pharmaceutical product development continues to gain momentum; this trend is due to the advantages of increased flexibility, speed of implementation and lower capital investment. In particular, they are seen as a means to accelerate the production of material for clinical trials. However, a primary concern regarding the use of such technologies is the impact and level of leachables in the final drug substance. Typically this concern is addressed through a risk assessment utilizing extractable…

The downstream purification process of any biologic has several objectives: purity, yield, and safety for humans or animals. A critical component of safety assurance is reducing virus to levels that meet stringent regulatory requirements. Virus reduction can be achieved through multiple complementary approaches and most processes rely on a combination of technologies that are designed primarily to achieve purification targets, but may also offer opportunities for virus reduction. The purpose of this project was to establish capabilities for producing a…

pDADMAC pretreatment of mammalian cell culture, coupled with Clarisolve® depth filters, provides for an effective pretreatment method by shifting the particle size distribution towards larger particles which can then be ‘matched’ to a graded depth filter designed for higher solids loading. Flexibility in filtration performance around pDADMAC dosing, flux rates, mixing hold times, and the variability in cell culture harvests provides for a robust clarification platform. Additionally, utilizing Clarisolve® filters for direct harvests, allows for a more compressed clarification train,…

Improvements in upstream process development often generate complex, high titer process streams, placing considerable demands on downstream processing steps. Protein aggregates in these feeds influence hydraulic performance of virus filters resulting in over-sized platforms and a significant impact on process economics. Virus filters from a broad range of manufactures provide robust viral clearance but the impact of aggregates on flux is dependent on the filter. The impact of conditioning protein solutions using prefiltration was assessed with several monoclonal antibody feed…

Current trends in the bioprocessing industry are driving mAb and plasma producers to formulate at higher protein concentrations. As a result, formulating using tangential flow filtration (TFF) may be limited in reaching these concentrations due to high pressures caused by highly viscous feed streams. Filtration devices used during processing have to be optimized in order to handle both high viscosity and pressures while maintaining high flux and excellent product recovery. In this study, a family of filtration devices was evaluated…

Continuous process optimization is a key factor in the biotech in-dustry. With higher and higher particle loads (>108 cells/ml), standard technologies for cell removal (midstream processing) – e.g. centrifugation, separation, membrane- and depth filtration – find their limits. The goal of the following study was to focus on making the midstream process more economical and investigate the efficiency increase for the cleaning of fermentation broths through alluvial filtration (see box) and its ease of scalability. This technology leads to a…

This poster highlights the Chromacassette® Technology: First practical modular chromatography platform. Supported-bed enables packing of any resin bead. Constant bed permeability, independent of resin compressibility even at V > 1000 cm/hr. Truly linear scalable. Eliminates packing methods and equipment. New operating domains and resin design are now possible for both bind and elute and flow-through. Hyper-Productive™ process enabling for single-use chromatography. PD devices will be available early 2018.

Four Protein A resins from leading industry vendors, including Amsphere™ A3, were screened for use in a capture step for a Herceptin biosimilar. Multiple parameters were assessed for their performance against the platform resin. Initially, a dynamic binding capacity (DBC) determination was made using a standard 4-minute residence time (RT) using purified protein. Performance of the resins were then screened for recovery and purity using harvested cell culture fluid (HCCF) where the MAb load was 80% of DBC at 10%…

Host cell proteins (HCPs) in biopharmaceuticals must be controlled in the downstream process (DSP) of antibody production, since it has critical influence on product quality. Generally, the majority of HCPs in harvested cell culture fluid are removed during the affinity step using Protein A chromatography resin, and the remaining HCPs are remove in the polishing step by a combination of ion-exchange chromatography, hydrophobic interaction chromatography, etc. Thus, identification of remaining HCPs after Protein A affinity chromatography process can work as…