Downstream

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…

Charge Heterogeneity Analysis in 10 Minutes

Platform methods, high resolution and ease of use have made the iCE system the gold standard for protein charge heterogeneity characterization for biopharmaceuticals. At 15-18 minutes per sample, iCE methods are already fast and simple but now they are even better. In this white paper we describe a high throughput 10 minute charge heterogeneity analysis method and performance improvements that provide automated analysis of 100 samples at a time.  

Purification of Oligonucleotides on TOYOPEARL GigaCap® Q-650S

TOYOPEARL GigaCap Q-650S is capable of delivering oligonucleotides of comparable purity and slightly higher process yields under the same operating conditions to those seen with resins requiring higher operating pressures. This capability allows chromatographers to purify oligonucleotides without the added expense of purchasing high pressure manufacturing equipment.

Polishing of Monoclonal Antibodies Using Capto™ adhere ImpRes in Bind and Elute Mode

MAbs and MAb conjugates are today in great demand for use as biopharamaceuticals. As a result, more cost-effective, efficient, and flexible process purification schemes are one of the highest priorities for MAb manufacturers. In this work, results from two case studies using Capto adhere ImpRes, a multimodal anion exchanger designed for polishing, are presented. Two different MAbs were purified in bind and elute mode. The results show high yields of MAb monomers, good clearance of aggregates, HCP, and leached protein…

Streamlining Downstream Process Development

Normal-flow filtration is used throughout downstream processes for biologics including depth, sterile, and viral filtration applications. Because of its ubiquity in large-scale biomanufacturing, using the most efficient normal-flow filter media area and type can lead to significant cost savings. But determining the most effective media type and area can be time consuming, labor intensive, and complicated because of a lack of specialized laboratory-scale equipment.

The equipment commonly used in this evaluation presents several disadvantages, including poor integration of multiple components from different sources (including pressure sensors, signal conditioning, data acquisition, balances, and pumps), continuous operator monitoring, and decreased portability of most systems due to their numerous components. This white paper describes how Genentech Inc. and PendoTECH collaborated on developing an integrated filter-sizing system that addresses these challenges and aids in efficient evaluation of filter media for manufacturing-scale processes.

A Novel Filtration System Used to Intensify Upstream and Downstream Operations

Want to improve your bioprocess? The ATF™ System from Refine Technology was designed to do just that. Used in various bioprocessing applications, this system provides a more reliable and efficient process of cell separation with the inherent ability to support cell growth to extreme concentrations. This educational webcast discusses the Alternating Tangential Flow technology of the ATF System and its many benefits, including:

• Near-linear scale-up
• A filtered product stream ready for purification
• Faster, simpler virus production with a reduced cost of goods
• And more

Join John Bonham-Carter, VP of Sales and Business Development at Refine Technology, as he provides a detailed look at the ATF System, its applications and its benefits.

pDNA Production and Purification

In recent years, there has been a growing demand for DNA-based therapeutics, commonly known as gene therapy, to combat genetically-based pathologies. Because plasmid DNA have significantly different biophysical characteristics than proteins, their production can have a drastic impact on downstream purification unit operations. Join Dr. Stephen Palmieri and Dr. Jesse McCool of Lonza Biologics and Dr. Fred Blattner of Scarab Genomics as they take a detailed look at the challenges faced by pDNA manufacturers and how Lonza’s pDNA Production Platform addresses those challenges.