January 2014

Happy 2014! Here is how the new year is shaping up for us so far. Our rotating editorial themes will remain much the same — but our manufacturing theme now shares the spotlight with increased visibility of analytical discussions. Our senior technical editor, Cheryl Scott, has highlighted specific methods in each issue of 2013 and will continue that throughout this year as well. For a list of her 2014 topics, take a look at our editorial calendar online at http://advertising.bioprocessintl.com/editorial-calendar (where you can also find out about our upcoming supplements and special reports). Each BPI Lab installment includes citations of past BPI articles in which the “method of the month” has been applied. Similar to our manufacturing theme that incorporates overarching quality functions as well as hands-on work in formulation and fill–finish, work with analytical methods is for many of you both outside of and integral to your day-to-day tasks. We realize the need to address such technologies as topics...

The following is a report from a workshop on innovation in biopharmaceutical manufacturing held at the Annual bioProcessUK Conference in Bristol on 29 November 2012. The aim of the workshop was to access the experience of practitioners in the United Kingdom so as to understand better the challenges and opportunities for innovation in this sector. The workshop addressed the drivers that influence the implementation of process improvements and novel technologies in biopharmaceutical manufacture from the perspective of both manufacturers and vendors. Biopharmaceutical companies must continuously improve and innovate to remain competitive and respond effectively to future challenges. However, the desire for modern manufacturing processes must be balanced against potential benefits, regulatory constraints, and risks. The Engineering and Physical Research Council (EPSRC) created the EPSRC Centre for Innovative Manufacturing in Emergent Macromolecular Therapies in 2011 with remit to create biomanufacturing innov...

The 2013 biennial meeting of the European Society for Animal Cell Technology (ESACT) was in Lille, France this past June. While there, BPI editorial advisor Miriam Monge (vice president of Biopharm Services Ltd.) interviewed ESACT executive committee member Hitto Kaufmann, PhD (vice president of biopharmaceutical process sciences for Boehringer Ingelheim). They talked about some scientific developments being discussed at this year’s ESACT conference as well as Boehringer Ingelheim’s recent announcement about setting up in China and Kaufmann’s own thoughts on the key future trends for biopharmaceutical development and manufacture. Hitto Kaufmann is currently heading a department that develops drug substance and drug product manufacturing processes and operates a clinical supply facility including fill and finish. Previously he was responsible for upstream development at BI’s Biberach site as director of upstream development and later as global head of mammalian cell culture processes. He joined Boehringer ...

An early BPI Lab article addressed the power of liquid chromatographic separations for biopharmaceutical laboratory use ( 1 ). Such techniques separate biomolecules based on a number of different properties: size, solubility, hydrophobicity/-philicity, binding affinity. The next most powerful means of separation — and thus high-resolution identification — of nucleic acids and proteins/peptides is based primarily on electrostatic properties: electrophoresis. Although it doesn’t really work in a process or preparative setting, it is a fundamental technique in modern biopharmaceutical laboratories, where it has become a relatively inexpensive, uncomplicated, and fast analytical method. Electrophoresis is often used for monitoring the progress of other protocols. Techniques such as isoelectric focusing (IEF) offer high-resolution separation of proteins based on their charge differences. Size separation is primary in sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Combining the two for “t...

For most biopharmaceuticals, potency is assessed in a bioassay by comparing dose–response curves of the test material and a reference standard. As with all analytical techniques, such assays require criteria by which their execution can be judged objectively to be valid, regardless of whether the desired or expected result is obtained for the test sample. PRODUCT FOCUS: BIOLOGICS PROCESSFOCUS: R&D, QC WHO SHOULD READ: PRODUCT AND PROCESS DEVELOPMENT, ANALYTICAL, QC KEYWORDS: IMMUNOASSAYS, POTENCY ASSAYS, PRODUCT RELEASE, REFERENCE STANDARDS, CONTROL SAMPLES, SAMPLE ACCEPTANCE LEVEL: ADVANCED The purpose of this paper is to provide guidance on setting assay acceptance criteria (AAC) for potency assays based on multiwell plates. Multiple components of the overall assay system — from instruments to incubation media — need to be within defined limits to permit execution of a valid assay, so they are tested for suitability either before or during the assay. Because of the complexity of bioassay systems, n...

Transgenic plants are increasingly considered a competing system for producing high-value recombinant proteins for biomedical and industrial purposes at affordable costs ( 1 ). Researchers have shown that molecular farming (or biopharming ) is a secure technology that is capable of rendering valuable recombinant proteins free of toxins and animal pathogens in a relatively short time ( 2 , 3 , 4 , 5 , 6 ). Scientists have also demonstrated that most recombinant antibodies produced in plants maintain their functional properties (substantial bioequivalence) as well as do those produced in mammalian cell cultures ( 7 , 8 ). Full antibody 14D9 is an IgG1- type immunoglobulin from mice that catalyzes the enantioselective protonation of prochiral enol ethers ( 9 ). Recombinant full antibody r14D9 has been expressed in Nicotiana tabacum plants as well as organ, tissue, and cell-suspension cultures. Highest yields were obtained with the KDEL retrieving signal to the endoplasmic reticulum (Ab-KDEL line) ( 10 ,

The biopharmaceutical industry is targeting high-concentration protein formulations to enable subcutaneous administrations. Such administration can provide better patient convenience than intravenous administration. One challenge associated with high-concentration formulations is increased electrostatic interaction between proteins and excipients. That is a result of increased protein-charge density at high protein concentrations. Such interactions can create an offset between excipient levels in final products and diafiltration buffers in ultrafiltration processes. The effect of such electrostatic interactions in a membrane process is known as the Donnan effect . The Donnan effect on excipient levels has received significant attention in recent years. Theoretical modeling has been developed to predict excipient and pH changes as a result of the Donnan effect in monoclonal antibody (MAb) processes. One model based on the Poisson–Boltzmann equation provided good prediction of excipient levels in the final...

When designing a recombinant protein production process, a high number of parallel cultivations must be carried out. That task is typically performed using batch cultures in shake flasks or microwell plates, in which fermentation conditions are not monitored. To overcome that limitation, we combined the SensorDish Reader and Shake Flask Reader systems (from PreSens) with an enzymatic glucose delivery system (EnBase technology from BioSilta Oy) for Escherichia coli cultivations. Our objective was to determine whether SensorDish reader cultures would yield reproducible process data that were directly comparable to those at shake flask scale (Photo 1). Photo 1:  Especially in the biopharmaceutical industry, a consistent process development from microliter to industrial scale requires consideration of many different perspectives to completely understand a transition process. We created a design of experiments (DoE) to determine optimal parameters at two different cultivation scales. Such batch-type cultivat...