Downstream Processing

Sartobind® pico and Sartobind® nano on Akta® – Optimizing Conditions for Liquid Chromatography Systems

Sartobind pico and Sartobind nano are the laboratory-scale mem-bers of the Sartobind membrane adsorber family. Accordingly, their small size and high throughput make them perfect for use on automated chromatography systems such as the popular Akta series offered by GE Healthcare Life Sciences.While both the pico and nano capsules are designed for high flow rates, the array of valves, pumps, monitors and the flow restrictor found in a typical flow path for gel chromatography can cause unacceptable pressures at the…

Aggregate Removal from Monoclonal Antibody with Nuvia™ HR-S Media

Nuvia HR-S media is a new cation exchanger designed for high resolution of closely related product impurities such as aggregates. It delivers excellent resolution, with a final aggregate content of <0.3%, and high recovery of >80% from a heterogeneous feed of monoclonal antibody aggregates and monomer. Aggregate content and recovery in the eluate were shown to be functions of the target conductivity measured at the end of collection.

Simpler and More Efficient Viral Vaccine Manufacturing

Human and veterinary vaccines are divided into five main categories: conjugate, toxoid, subunit, inactivated (killed), and live (attenuated) vaccines (1). The vast majority of currently licensed human and veterinary vaccines are inactivated or live (2, 3). They are produced mostly using adherent cells: primary cells such as chicken embryo fibroblasts (CEF), human diploid cells such as MRC-5, or continuous cell lines such as Vero and MDCK (4). The pioneering legacy inherited by vaccine manufacturing development has led to strategies for…

Emerging Challenges to Protein A

Protein A affinity chromatography has been a target for replacement since its commercial debut, mainly because of its high acquisition cost. The technique became established despite the cost because it was born into an industrial culture that favored speed to market over manufacturing economy (1). Vendors have since strengthened protein A’s position with incremental but worthy improvements such as higher capacity, lower ligand leaching, and modest tolerance of NaOH. Collateral improvements in polishing technologies, such as the high throughput and…

Virus Risk Mitigation for Raw Materials

Recombinant protein–based medicinal products and modern cell-based vaccines have a very strong safety history with respect to viral and microbial contamination. However, virus contamination incidents do occur occasionally in manufacturing processes, and they can consume many resources and be expensive to rectify. The root cause of contamination incidents in recent years is most likely the use of contaminated raw materials. These include bovine serum contaminated with reovirus, epizootic hemorrhagic disease virus, Cache valley virus or vesivirus 2117; porcine trypsin contaminated…

Development Strategies for Novel Vaccines for Infectious Diseases

In a vaccine development program, the probability of success at each transition decreases, even though the actual probability of moving from one phase to another can be 50–80% (Figure 1). Many compounds and vaccine candidates are screened out even before they get into preclinical studies. Developers can implement different approaches to reduce product failure risk before a program gets expensive, including Establishing a product development plan (PDP) Identifying and mitigating risk with gap analysis Learning from the mistakes of others…

Industry Adoption of Membrane Adsorbers

Membrane adsorbers (MAs) are the fastest-growing segment in single-use bioprocessing. But their future is not entirely certain. According to BioPlan Associates’ latest survey of biopharmaceutical manufacturing, the MA market has been growing at ~20% annually since 2006 (1). Paradoxically, however, the segment may not be a true “rising star.” Our study also shows that MAs remain among the least-often adopted devices among biomanufacturers. So the question of how and whether MA technology can revolutionize bioprocessing remains open. Market for Membrane…

A Platform Approach for the Purification of Domain Antibodies (Dabs)

A three-step purification process for a Dab successfully developed and verified. The step yield ranged between 86% and 99%, giving a total process yield of approximately 81%. The ECP in the start sample was more than 200 000 ppm whereas the final sample contained only 5.5 ppm. The endotoxin content in the feed was approximately 2 million endotoxin units/mg of protein and the final sample was below the limit of quantification. Protein L leakage was undetectable in all samples. This…

MFI for Particle Characterization of Biopharmaceuticals Today

Micro-flow imaging (MFI) is a sensitive, simple and automated method for the analysis of sub-visible particles and translucent protein aggregates that provides particle size, count and morphology. Because it quantifies particle shape, this technology can discriminate groups of particles from each other, and evaluate how these groups change over time. In this article, we provide an overview of how biopharma is using MFI to characterize and monitor their protein formulations in new and emerging applications.  

POROS® CaptureSelect™ Affinity Columns for Rapid, Small-Scale Purification and Sample Preparation of Recombinant Proteins

This application note describes a model system in which Enbrel® protein, a fusion protein consisting of TNF receptor and IgG1 Fc, is purified from a dilute sample matrix for further analytical characterization. The resulting purified protein was subjected to N-linked glycan structural analysis by mass spectrometry (MS) and its aggregation state was assessed by analytical ultracentrifugation (AUC). This data set serves as a model to demonstrate the capability of the POROS® CaptureSelect™ product line for the affinity purification of biomolecules…