Search Results for: design of experiments

New ambr® bioreactor systems enhanced with software for Design of Experiments (DoE)

Sartorius Stedim Biotech (SSB) today announced the 2015 version of the ambr® systems (ambr® 15 and ambr® 250) will be supplied with integrated BioPAT® MODDE Software for Design of Experiments (DoE), powered by Umetrics. This will allow bioprocess scientists to easily implement DoE into their work flow for simpler process optimisation and scale-up to larger single-use BIOSTAT® pilot and manufacturing scale bioreactors, making bioprocess development faster and more cost-effective. The integrated DoE software will enable scientists to quickly establish a…

Ask the Expert – Optimizing Cell Culture Media Supplements: Using Design of Experiments for CHO and HEK293 Cells

with Francesc Gòdia (chemical engineering professor at the Universitat Autònoma de Barcelona) Chemically defined media free of animal-derived components are required for bioprocess operations based on mammalian cells. Supplementation of commercial media with specific compounds was studied using a design of experiments (DoE) approach to screen the most efficient compounds for two cell lines and then determine their optimal conditions. DoE allowed simultaneous testing of several compounds to determine potential interactions among them in addition to their individual effects. Gòdia’s…

Optimization of HEK 293 and CHO-S Cell Growth by Supplementation of Non-Animal Derived Components Using Design of Experiments (DoE)

Mammalian cells are a widely used expression platform for the production of recombinant therapeutic proteins or viral particle-based vaccines since they typically perform appropriate protein post-translational modifications and authentic viral particle assembly. Of the available mammalian cells, CHO and HEK 293 are some of the most industrially relevant cell lines because they are cGMP compliant and are able to grow in suspension in a variety of serum-free media. Of note, production of human therapeutics in mammalian cell culture has become…

Design of Experiments with Small-Scale Bioreactor Systems: Efficient Bioprocess Development and Optimization

Design of experiments (DoE) is one of the most valuable techniques for organized and efficient planning, execution, and statistical evaluation of experiments. Although a DoE investigation can be completed using several runs in one bioreactor, small-scale bioreactor systems designed for parallel operation (such as the ambr15 or ambr250 systems) provide the optimal basis to economically realize a series of experiments. Because of the multitude of interdependent parameters involved in applications such as cell line development, culture media screening, and the…

Design of Experiments for Fed-Batch Process Development in Shaken Cultures

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…

Biological Assay Qualification Using Design of Experiments

In 2012, the United States Pharmacopeia (USP) published a complementary set of three guidance documents on the development, analysis, and validation of biological assays (1,2,3). USP chapter recommends a novel, systematic approach for bioassay validation using design of experiments (DoE) that incorporates robustness of critical parameters (2). Use of DoE to establish robustness has been reported (4,–5), but to our knowledge its use in qualification or validation protocols for assessing assay accuracy, precision, and linearity is not described in literature.…

Think Strategically for Design of Experiments Success

Global competition fueled by the power of information technology has forced the pharmaceutical and biotechnology industries to seek new ways to compete. The US Food and Drug Administration (FDA) has promoted quality by design (QbD) as an effective approach to speed up product and process development and create manufacturing processes that produce high-quality products that are safe and effective (1,2,3). Statistical design of experiments (DoE) is a tool that is central to QbD and the development of product and process…

Optimization of a PEGylation Reaction Using Design of Experiments (DOE)

The covalent attachment of polyethylene glycol (PEG) to a therapeutic protein is frequently used to increase the half-life of that protein in patients while reducing their immunogenic response. The PEGylation reaction can be optimized to maximize the desired product (average moles of PEG per mole of protein) molecules while limiting undesired byproducts (under- and over-PEGylated species). In the example discussed here, the desired molecule was mono-PEGylated, yet the preoptimized reaction yielded multi-PEGylated species visible by SDS-PAGE. The goal of our…

Ask the Expert: Leveraging Cloud Capabilities to Execute CHO Cell Culture Experiments

Process development teams have relied for decades on benchtop bioreactors to perform mammalian-cell–culture experiments; however, doing so requires significant resources and staffing. During a 24 February 2021 presentation, Deborah Pascoe, PhD, vice president of operations at Culture Biosciences (Culture), explained how to leverage her company’s high-throughput, cloud-connected bioreactors to execute Chinese hamster ovary (CHO) cell cultures with high reproducibility and scalability. Pascoe’s Presentation Pascoe described her company as an extension of a customer’s laboratory. After technology transfer, Culture and a…