Eppendorf

Truly Parallel – Microbiology

DASGIP® Bioreactor Systems for Microbiology.

Truly Parallel – Cell Culture

DASGIP® Bioreactor Systems for Cell Culture Applications.

Product Brochure: Most Compact

DASbox® Mini Bioreactor System for Cell Culture and Microbiology.

Product Brochure: The Smarter Solution

BioFlo® 320 bioprocess control station.

High Cell Density Fermentation of Escherichia coli Using the New Brunswick™ BioFlo® 115

This application note presents a successful example of a high density fermentation of Escherichia coli (E. coli) using the New Brunswick BioFlo 115 benchtop, autoclavable fermentor from Eppendorf. The highest optical density (OD600) achieved in this study was 140 at 11 hours (h) without optimized fermentation medium and conditions.

Evaluation of the BioFlo® 320 Process Capabilities

Taking the Strain

Today, the rapidly expanding demand for vaccine products for viral diseases such as rabies has necessitated the development of more sophisticated production techniques based around cell culture systems. This article reviews vaccine production strategies, with a focus on rabies, looking specifically at the use of the Vero cell line - used worldwide and approved by the US Food and Drug Administration - as well as media technology and the bioreactor options available.

Growing Potential: mAb Production With Fibra-Cel

The market for monoclonal antibodies continues to grow steadily, but there are still bottlenecks in the development of new products, including long development times mostly due to R&D. Advanced bioprocess equipment that meets the specific demands of mAb-producing hybridoma can accelerate design and production, and reduce overall development costs along the way.

DoE Bioprocess Development

Eppendorf DASware is the effective route to design of experiments (DoE) in early-stage bioprocess development. Claudia M Huether-Franken and Sebastian Kleebank explain how the design software applies DoE to DASGIP parallel bioreactor systems.

Product Brochure: Bacteria Welcome

BioBLU® f – Single-use vessels that meet microbial demands.

Parallel Cultivation of Microorganisms Using Rigid-Wall Single-Use Bioreactors

In recent years, single-use bioreactor solutions have been successfully established for animal and human cell culture processes. Such technology is under investigation for microbial applications. Sebastian Kleebank is development and application laboratory manager for the Eppendorf line of bioprocess products. His cell culture and microbial laboratory performs parallel cultivations for prototype testing, user training, and software development.

Advanced Biofuel Development Using Parallel Bioreactor Systems

To make biofuels such as bioethanol and biodiesel competitive with fossil fuels extensive research in the field of process development is needed. Taking into account the special requirements of biofuel production, such as high temperatures, multi-step procedures and anaerobic conditions, effective tools for streamlining development efforts must be provided.

Product Brochure: When Less is More

New Brunswick’s CelliGen® BLU Single-Use Bioreactor System.

Product Brochure: Extra Versatile

New Brunswick’s BioFlo®/CelliGen® 115 Autoclavable Fermentor and Bioreactor handles a wide variety of cells to meet your every need.

Product Brochure: Customizable Control

New Brunswick™ BioFlo® 510 benchtop SIP fermentation systems.

Product Brochure: Benchtop Convenience

New Brunswick™ BioFlo® 415 SIP fermentation systems.

Advanced Control

New Brunswick™ BioFlo® 310 2.5 L - 14.0 L Fermentation Systems

Product Brochure: Compact Mobility

New Brunswick™ BioFlo® 610 mobile SIP fermentation systems.

Product Brochure: Scale-Up Seamlessly

New Brunswick™ BioFlo® Pro pilot to production SIP fermentation systems.

Seamless

New Brunswick™ BioCommand® Software for Fermentation & Cell Culture

High Cell Density E. coli Fermentation using DASGIP® Parallel Bioreactor Systems

In contract manufacturing one of the most challenging needs is the adaption of proprietary production processes, their optimization as well as process improvements after a transfer from one Contract Manufacturing Organization (CMO) to another. Each single process modification has to be verified with comparable data. The following application note gives an example for such an adapting procedure achieved by the use of the DASGIP Parallel Bioreactor System. The established process of one CMO included a high cell density fed-batch process, controlled by the key process parameters pH, agitation, temperature and growth dependent glucose feeding. This process was adapted and optimized for the use with another CMO.

Scalability of Parallel E. coli Fermentations in BioBLU® f Singleuse Bioreactors

Single-use bioreactor solutions have been successfully established in animal and human cell culture in the last years. Now this technology is going to make its way for microbial applications. In the following case study reproducible process control was achieved with single-use mini bioreactors and 1 L single-use vessels running in parallel. Fermentation of E. coli K12 led to highly reproduclible results thus proving the tested rigid wall single-use stirred-tank vessels to be an appropriate tool to accelerate microbial process development and shorten time-to-market in biopharmaceutical industry.

Scalable Expansion of Human Pluripotent Stem Cells in Eppendorf BioBLU® 0.3 Single-Use Bioreactors

The routine application of human pluripotent stem cells and their derivatives in regenerative medicine and innovative drug discovery will require the constant supply of high cell numbers in consistent, high quality. Well monitored and controlled stirred-tank bioreactors represent suitable systems to establish up-scalable bioprocesses enabling the required cell production. The following application note describes the successful cultivation of human pluripotent stem cells in suspension culture using Eppendorf BioBLU 0.3 Single-use Vessels in a DASbox® Parallel Mini Bioreactor System.

Cultivation of Human CAP® Cells: Evaluation of Scale-Down Capabilities using Single-Use Bioreactors

Increasing process complexity coupled with rising cost pressures and rapidly evolving regulatory requirements makes today’s process development efforts a special challenge. The pressure of achieving faster time-to-market for new and innovative biotechnological products has led to the need to optimize every element of the total development workflow. The following application note illustrates how the DASbox ® Mini Bioreactor System combined with the BioBLU® 0.3c single-use vessels supports bioprocess development in human cell culture. Scale-down capabilities were investigated by comparison of 500 mL cultures in a DASGIP® Parallel Bioreactor System with 170 mL cultures in the DASbox using the BioBLU 0.3c single-use vessel.

Hybridoma culture using Eppendorf New Brunswick™ CelliGen® 310 with Packed-bed Fibra-Cel® Basket Impeller

The Fibra-Cel packed-bed basket technology has been established as an excellent method for the growth of suspension and anchorage-dependent cell lines. The three dimensional structure of the Fibra-Cel disk provides an excellent solid-support matrix for the entrapment or attachment of animal cells, allowing constant perfusion of nutrients in a low-shear environment. In this application note, we show that Hybridoma cells can be successfully cultivated in high densities in the 2.5 L packed-bed Fibra-Cel basket controlled by a CelliGen 310 bioreactor.

CHO Cell Culture with Single-Use CelliGen® BLU Packed-Bed Fibra-Cel® Baske

The packed-bed basket technology, developed by New Brunswick Scientific (acquired by Eppendorf Inc. in 2007), provides a shear free environment for production of animal cells. At present, little information is available on the utility of the New Brunswick CelliGen® BLU singleuse bioreactor system for the production of secreted proteins, especially in perfusion mode of operation. Thus, this study was conducted to measure the growth and productivity of alkaline phosphatase (ALKP)-secreting rCHO.

Large-scale Production of Human Mesenchymal Stem Cells in BioBLU® 5c Single-use Vessels

Stem cell-based regenerative medicine has great potential to revolutionize human disease treatments. Among the various stem cell platforms, mesenchymal stem cells (MSCs) represent one of the highest potentials as evidenced by clinical trial activities. Currently, there are over 400 clinical trials based on MSCs registered at clinicaltrials.gov. Although successful expansion of MSCs in vitro has been well established, the large clinical-scale production of MSCs remains a bottleneck, potentially limiting the immediate clinical applications should some of the ongoing trials receive FDA approval. In this study, we demonstrate the success of large clinical-scale culture of human adipose-derived mesenchymal stem cells (AdMSCs) in an industrial single-use vessel at 3.75 liter (L) scale (working volume).

Single-Use Scalability: CHO Cell Culture using 5 to 50 L New Brunswick™ CelliGen® BLU Benchtop, Stirred-Tank Bioreactors

This study illustrates a protocol for the scale up of CHO cells using New Brunswick™ CelliGen BLU stirred-tank bioreactors equipped with 5-Liter (L) and 50-Liter (L) single-use vessels. CelliGen BLU is a versatile benchtop system for the culture of a variety of cell lines. This bioreactor has been designed to provide research and production facilities with single-use vessels that combine the benefits of both traditional stirred-tank design and single-use technology, capable of seamless process scaleup. Eppendorf has recently launched the CelliGen BLU 50 L system to address larger volume batch demands.

Insect Cell Culture Using the New Brunswick™ BioFlo®/CelliGen® 115 Benchtop Fermentor/Bioreactor with Spin Filter Assembly

This application report presents a simple protocol for achieving high-density culture of Spodoptera frugiperda (Sf9) cells using a New Brunswick bench-top, autoclavable stirred-tank reactor with a spin-filter assembly. Factors such as substrate concentration and metabolite buildup can be limiting for culture growth and viability at high densities. Using the spin filter in a 3.0 L vessel (2.0L working volume) attached to the BioFlo®/CelliGen® 115 cabinet, a cell density of 18.24 x 106 cells with viability over 90 % was achieved, outperforming the batch or fedbatch process.

Introduction to Pichia Pastoris in a Stirred-Tank Fermentor

This Applications Report presents a simple protocol for achieving high-density culture of Pichia pastoris (P. pastoris) cells using a New Brunswick benchtop, autoclavable stirred-tank fermentor or bioreactor.

Pitched-Blade vs. Spin Filter vs. Packed-bed Basket:CHO Cell Culture Comparison

In the following application note, the pitched-blade impeller, the spin filter impeller and the packed-bed basket impeller are discussed, highlighting the uses and advantages for each type. Then examples of actual CHO cell cultures are given for each impeller type; showing the perfusion capability when using the spin filter or packedbed basket impeller and the resulting higher cell densities over the pitched-blade impeller.

Fed-Batch Biofuel Production Process Using a New Brunswick™ BioFlo® 115

In Brazil, it is common in the biofuel industry to utilize a biochemical process in which glucose, fructose and sucrose (derived from sugar cane juice and sugar cane molasses) are used to produce biofuel through a fed-batch fermentation process. In this experiment, Saccharomyces cerevisiae is used for biofuel production from sugar cane juice. The fermentation process metabolizes glucose into ethanol, and is used to produce many biofuel products in large production volumes. This strain of yeast is also widely used in other industrial applications to manufacture enzymes and proteins for beer, wine and bread. In this application note, we show that Saccharomyces cerevisiae can successfully be cultivated in high densities to convert sugar cane into biofuel using a BioFlo 115.

Using Redox Measurements to Control Anaerobic Yeast Fermentation in a New Brunswick™ BioFlo® 310 Fermentor

In this study, we used Saccharomyces cerevisiae to produce ethanol under anaerobic conditions in a BioFlo 310 fermentor. Initially, the yeast culture was conducted under aerobic conditions; after 24 hours, the culture was switched to anaerobic growth for the production of ethanol. During the fermentation process, redox potential was maintained at -180 mV using a redox sensor and gas flow control. Toward the end of fermentation, a significant amount of ethanol was produced and the cells retained maximum viability. Our study indicated that with a redox sensor, the BioFlo 310 fermentor was capable of maintaining anaerobic conditions, extremely low oxygen level, during the entire ermentation process.

Sf-9 Insect Cell Culture using a New Brunswick™ CelliGen® 310 Bioreactor: Using headspace air overlay for reduced dCO²

This study describes a simple procedure for improving insect cell yields in a benchtop cell culture bioreactor. Here, yields of Spodoptera frugiperda (Sf-9) cells were increased by nearly 29 % through monitoring dissolved carbon dioxide (dCO2) levels in the culture and adding air to the vessel headspace to reduce dCO2. The method can also be used to maximize yields in a wide range of mammalian cell types.

Which Impeller Is Right for Your Cell Line? A Guide to Impeller Selection for Stirred-Tank Bioreactors

When growing microbes or animal cells in a stirred-tank reactor, it is critical to choose the impeller type that is best suited to your process. Select the wrong impeller, and you could make chop suey of your filamentous fungi. Pick the right impeller, and you could greatly increase yields of your fussy mammalian cultures such as Chinese hamster ovary (CHO) and Vero kidney epithelial cells. With a wide range of impeller designs to choose from, how do you tell which is right for your application? Here we describe six commonly used fermentor and bioreactor impellers, explain how they work, and identify which may perform best for culturing certain animal, insect, plant, yeast, and bacterial cell lines.

Optimization of HEK293 Cell Culture in a New Brunswick™ CelliGen® 115 Bioreactor for the production of recombinant GPCR

Heterologous expression of membrane proteins remains a bottleneck for structural characterization by x-ray crystallography. Such proteins represent approximately 30 % of the proteome and are not sufficiently represented in the Protein Data Bank (PDB)[1]. G-protein-coupled receptors (GPCRs) are an area of particular interest as it is estimated that one third of current FDA-approved drugs act through this class of receptors.

An Update on the Advantages of Fibra-Cel® Disks for Cell Culture

With a renewed acceptance of perfusion processes in cell culture, both in the laboratory and for production applications[1], New Brunswick packed-bed bioreactors using Fibra-Cel disks are seeing an upsurge in interest. This application note examines Fibra-Cel technology and its many advantages, from increased secreted protein yields to labor savings in applications including rabies vaccine production and production of the first licensed gene therapy drug.

Growing CHO Cells in a New Brunswick™ CelliGen® BLU Benchtop, Stirred-Tank Bioreactor Using Single-Use Vessels

The study presents a typical protocol for the setup and operation of the Eppendorf New Brunswick CelliGen BLU single-use, stirred-tank bioreactor, a versatile new benchtop system for the culture of a wide range of mammalian cells. This bioreactor has been designed to provide research and production facilities with a single-use vessel which combines the benefi ts of both traditional stirred-tank design and single-use technology, capable of seamless process scale-up. The system can be operated in batch, fed-batch or continuous modes. A procedure for culturing Chinese Hamster Ovarian (CHO) cells in a 5.0 L vessel, using CD CHO serum-free medium in a batch culture is described.

Anaerobic yeast fermentation for the production of ethanol in a New Brunswick™ BioFlo® 310 fermentor

Whether used for research or production, the versatile New Brunswick BioFlo 310 fermentor from Eppendorf allows growth of a wide variety of aerobic and anaerobic microorganisms, including bacteria, plant, algae, fungi and yeast. Its advanced controller can regulate up to four vessels simultaneously, 120 process loops in all. Here we demonstrate one facet of its versatility -- a technique for inducing ethanol production in yeast, by switching from an aerobic growth phase to an anaerobic steady-state culture.

Amino Acid Fermentation: Evaluation of Scale-Down Capabilities Using DASbox® Mini Bioreactors

Optimization of bioprocesses needs accurate monitoring and control while small working volumes are saving media and other resources. Evonik® has established a 2 L process for production of a nutritionally relevant amino acid in E. coli. The following application note descripes how this process was adapted to the smaller working volume of an Eppendorf DASbox Mini Bioreactor System. Multiple runs were performed with close monitoring of all relevant process parameters and comprehensive evaluation of data to prove reliable and reproducible results. The scale-down capabilities of the DASbox system were evaluated by comparing the fermentation results to the data collected at 2 L scale.

A Comparative Study: Small Scale E. coli Cultivation Using BioBLU® Single-Use and Reusable Vessels

In recent years single-use bioreactors gained more and more importance in animal and human cell culture. With the new line of BioBLU f rigid wall, stirred-tank single-use vessels Eppendorf offers premium solutions for microbial applications.

Open Interface

DASware® migrate: Manage your bioprocess data with DASGIP®.

Dependable Design

New Brunswick™ CelliGen® Pro pilot to production SIP bioreactor systems.

System Flexibility

New Brunswick™ CelliGen® 510 benchtop SIP bioreactor systems.

Controlled Cultivation of Stem Cells – Factors to Consider When Thinking of Scale-Up

Great hopes and expectations are linked to stem cells as a tool for drug discovery and to stem cell-derived products in therapeutic applications. Though several products have made it to commercial stage, most of the research is still performed in small scales using simple cultivation systems such as spinner flasks or T-flasks.