Downstream

Best Practices For Successful Filter Integrity Testing Using The Water Intrusion Test (WIT) Method

Hydrophobic filter cartridges which utilize sterilizing-grade polyvinylidene fluoride or polytetrafluoroethylene membranes are widely used in the pharmaceutical industry to sterilize compressed air or gases for product contact, as well as acting as sterile tank vents. However, hydrophobic membranes can also make filter integrity testing in situ difficult, as the membrane must be fully wetted prior to testing by the widely accepted integrity test methods of Bubble Point and Forward Flow. The Water Intrusion Test (WIT) is widely accepted within the…

CaPure-HA™: Purification of MAb Fragments

Making use of chromatography resins with better selectivity, resolution and capacity is one approach to solving the problem not only of aggregate removal, but also that of MAb fragment and other digest products, in a column chromatography purification step. The data presented here demonstrate the capabilities of CaPure-HA to separate Fab and Fc fragments from undigested MAb in a papain digest of an IgG1 monoclonal antibody.

Cadence™ Single-Pass TFF Coupled with Chromatography Steps Enables Continuous Bioprocessing While Reducing Processing Times and Volumes

Continuous bioprocessing initiatives have accelerated in recent years. The transition from batch to continuous bioprocessing offers numerous advantages, including process time savings and lower capital costs due to smaller equipment, tanks, and tubing sizes. Additionally, hold tanks may no longer be needed between unit operations, which can be especially valuable for facilities with limited manufacturing floor space. Furthermore, reduced system hold-up volumes improve product recoveries and also contribute to the smaller system footprint of continuous bioprocessing systems. One unit of…

Volume Reduction and Process Optimization with Cadence™ Inline Concentrator

Pall’s Cadence™ single-pass tangential flow filtration (SPTFF) technology provides a simple solution for continuous concentration and optimization of Downstream Processing (DSP) steps. It eliminates the conventional TFF recirculation loop and allows product to be concentrated in a single pump pass. The single-pass operation eliminates any mixing or foaming issues and exposes the product to low shear; thus, it is optimal for the processing of fragile and shear-sensitive molecules as well. The single-pass feed flow rates are lower compared to conventional…

CaPure-HA™: DoE Optimization of Elution Conditions

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. The data presented here demonstrate the optimization of the CaPure-HA elution using a DoE approach.

Pressure Interruptions (Stop/Start) During Virus Filtration: Assuring Safety Using Robust Process Technology and An Appropriate Risk Mitigation Strategy

Following publications discussing the impact of pressure interruptions on the retention levels of virus filters, an increased focus is being placed on filter users’ own risk mitigation strategies. Users are increasingly requesting technical guidance and support with viral validation from filtration experts, as part of their overall product support package. This guidance document provides a high level of assurance to filter users by demonstrating how combining robust virus clearance technology with the use of standard operating procedures can assure a…

Sterile Vent Filtration on Ozonated Water Tanks

With many pharmaceutical processes requiring large volumes of water, it is critical that any pharmaceutical-grade water used is protected from particulate or microorganism contamination, in order to ensure that process operations do not become inadvertently contaminated. Several approaches can be used to ensure that water remains free from contamination, including the storage of purified water or WFI (Water For Injection) at a minimum temperature of 80°C to discourage microbial growth in the storage system. Another approach is to add ozone,…

Computer-Aided Assay Development for Charge Heterogeneity Analysis by iCE

Unlike chemically synthesized drugs, protein therapeutics are a dynamic heterogeneous mix of active compounds. Due to their complexity, analytical techniques like isoelectric focusing have become indispensable tools in evaluating biologic preparations. The resulting surge in charge isoform analysis has led to major advances in instrumentation, such as Imaged Capillary Electrophoresis (iCE™)2 . However, to obtain the full benefit from improved instrumentation requires the coinciding development of robust assays. The goal of this note is to promote the successful application of…

Use of ScreenExpert RoboColumns for High Throughput Study of Loading Conditions on HyperCel™ STAR AX and MEP HyperCel Sorbents for MAb Purification in Flow-Through Mode

The use of HyperCel STAR AX and MEP HyperCel sorbents for MAb polishing provides efficient contaminant removal. A study to screen loading conditions for MAb purification in flow-through mode on MEP HyperCel and HyperCel STAR AX sorbents was conducted using 200 μL ScreenExpert RoboColumns. The study demonstrates the suitability of ScreenExpert RoboColumns for scale down evaluation of chromatographic parameters, and the good consistency with data obtained on 1 mL PRC prepacked columns. ScreenExpert RoboColumns allow loading of high sample volume…

Implementation of Mustang® Q Membrane Chromatography as a Polishing Step (Residual DNA Removal) in Monoclonal IgG1 Production from CHO Cell Culture

In collaboration with ProBioGen AG, Berlin, Germany, Mustang Q membrane chromatography was evaluated as a polishing step following protein A affinity and cation exchange chromatography to remove residual host cell DNA during a Monoclonal antibody (MAb) purification process at 250 L cell culture production-scale. Data indicated efficient DNA clearance by the Mustang Q membrane chromatography step during the process (96%). Additionally, contribution of the Mustang Q membrane chromatography polishing step to HCP removal was shown. Based on a virus-spiking scale-down…