2015

Improved Fluorescent Labeling Efficiency of N-Linked, High-Mannose Oligosaccharides: Using 8-Aminopyrene-1,3,6-Trisulfonic Acid (APTS) for Analysis of Glycoproteins

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Glycosylation of proteins, including monoclonal antibodies (MAbs), is recognized as important for the efficacy, immunogenicity, antibody-dependent cell-mediated cytotoxicity (ADCC), and complement-dependent cytotoxicity (CDC) of biotherapeutics (1–6). So research and development of protein candidates is increasingly focused on the effects of glycosylation and how its pathway is affected in the Golgi system of cells involved in biosynthetic processes (7). Such attention on glycosylation has helped advance analytical technologies such as high-pH anion-exchange chromatography (HPAEC) (8); normal-phase chromatography (NP- HPLC), hydrophilic-interaction chromatography…

March 10, 2015

The Importance of the Concentration-Temperature-Viscosity Relationship for the Development of Biologics

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JIM DELILLO (WWW.FREEIMAGES.COM) Patient preference and a competitive landscape in the parenteral market have fueled the need for convenient delivery systems and a desire for less‑frequent dosing injections. Monoclonal antibodies (MAbs) often have high dose requirements, so they must be formulated at very high concentrations (1). At low concentrations, an antibody solution’s viscosity increases moderately as a function of protein concentration. But at high concentrations (>100 mg/ mL, depending on the molecule), viscosity increases exponentially (2, 3). Thus, a specification…

March 10, 2015

Characterization of Postcapture Impurity Removal Across an Adsorptive Depth Filter

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In the manufacture of monoclonal antibodies (MAbs), the first purification step following harvest clarification is normally protein A affinity chromatography because of its high selectivity for IgG and high process yield (1, 2). At this stage, a MAb is eluted from a protein A ligand at low pH and then held or adjusted to a low pH (pH ≤ 3.8) for a given amount of time before pH adjustment, usually ≥30 minutes, in a virus inactivation (VI) step targeted at…

March 10, 2015

A Multidisciplinary Approach to Manufacturing Biotherapeutics

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Optimizing antibody manufacturing processes has gone beyond the first-order goal of achieving elevated protein titers and now also focuses on understanding biologic and manufacturing process variables that define cellular machinery and protein quality. A holistic approach to biotherapeutic manufacturing incorporates several applied disciplines such as biology, engineering, process control, signal processing, and modeling to reduce the “black-box†model of cell- based protein production into an operational design space. This is in line with the US Food and Drug Administration’s quality…

March 10, 2015

A Quick Guide for Sourcing Biopharmaceutical Raw Materials

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Before the ratification of regulatory guidelines from The International Conference on Harmonisation of Technical Requirements for Registration of Pharmaceuticals for Human Use (ICH) Q8–Q11 (1–4) — whose scope includes raw materials for biopharmaceutical production — many drug manufacturers chose the most cost-effective and readily available raw materials sourcing options without specifically considering the provenance of those materials. Depending on the chosen supply chain, such materials could be of widely varying quality and not necessarily suitable for a destined application. Raw-material…

March 10, 2015

Ask the Expert – 2D DIGE Western Blots for HCP Detection and Coverage Determination: Advantages of Fluorescence over Colorimetric Detection

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with Jamina Fiedler, BioGenes GmbH Biopharmaceutical products must be free of by-products (such as host-cell proteins, HCPs) for approval by authorities such as the US FDA. For that, a clear proof is required of sufficient HCP detection by the HCP assay used. The two-dimensional (2D) DIGE Western blot method is the standard technique for detection of HCP coverage. Colorimetric detection has several limitations that can be overcome by the use of fluorescent detection. Jamina’s Presentation Practical differences between 2D DIGE…

March 10, 2015

From the Editor

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In this month’s lineup, you’ll find a common theme of balancing operating costs against company cultures and corporate philosophies. This is far from new: You can’t responsibly run a business without making such calculations! But the value in taking a fresh look at them lies in how we are forced to dust off and reassess some assumptions (even prejudices). It is easy (and all too human) to persist with outmoded assumptions — and we cannot afford to do that given…

February 6, 2015

Spotlight February 2015

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Niche-Disease: Amyloidosis by Cheryl Scott Amyloidosis is a family of rare diseases involving a build- up of misfolded (and thus insoluble) amyloid protein in one or more organs. The condition affects different organs in different people, involving different types of amyloid protein. It most frequently affects the heart, kidneys, liver, spleen, nervous system, and digestive tract. Severe amyloidosis can cause life-threatening organ failure, and there is no cure. Some 60 amyloid proteins have been identified, with about half of them…

February 6, 2015

30 Years of Upstream Productivity Improvements

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We recently completed an analysis of the past 30 years of industry progress in commercial-scale expression titers and bioprocessing yields. These basic measures of biopharmaceutical manufacturing efficiency also benchmark the technological progress made in bioprocessing over recent decades. Titer and yield improvements generally indicate related bioprocessing cost savings, something most commercial-scale manufacturers work to improve. This focus on efficiency and productivity has led to constant bioprocessing improvements even for long-approved and -marketed products. Our findings indicate that although upstream titers…

February 6, 2015

A Case for Biopharmaceutical Operations in Low-Cost Countries to Maximize Return on Investment

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A company’s success is associated with its growth. An expanding company hires more employees (full-time equivalents, FTEs) to support its commercial products, to develop new products, and to provide administrative services. As it grows, an organization can become more complex and its operations less efficient. Analysis of 2013 financial data showed that revenue of US biopharmaceutical companies increases with a growing number of FTEs at a slower rate than does cost (1). That study assumed that all US biopharmaceutical companies…

February 6, 2015