Thursday, May 16, 2019 from 9:00 AM to 5:00 PM EDT
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BWI Aiport Marriott - Baltimore, MD 
1743 W. Nursery Road
Linthicum Heights, MD 21090

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Carol Stanton 
Eppendorf Americas 

Cell Culture Technology

Scalability, perfusion and vaccine solutions

Complimentary workshop hosted by Eppendorf to discuss the latest developments in upstream bioprocessing methods.

Presenter Topics

  • Insights on vaccine development and current technology
  • Studies illustrating cell culture scale-up including lentiviral vectors and rAAVs
  • Various techniques of high density perfusion cell culture
  • Techniques for scaling bench to production
  • Small-scale perfusion using an ATF cell retention device with parallel bioreactor system
  • Advancing viral vector and vaccine process characterization with adherent cells using an integrated perfusion platform presented by Corning
  • Simplify and intensify cell culture with XCell™ ATF presented by Repligen®

More information:  cellexperts@eppendorf.com

Key Learning Objectives

  • Learn about technologies in cell culture upstream bioprocess development.  Focus will be on perfusion technologies and scale-up.
  • Get first-hand insights from academia and industry peers
  • Interact with other scientists and process engineers and grow your network


Thought Leadership – Insights on Vaccine Development and Current Technology
Driven by epidemic events and by governmental vaccination programs, there is a rising demand for development of new vaccines and the industry is growing at a double-digit rate. Manufacturing companies need to set up flexible processes, optimized to balance costs and time-to-market. New technologies open up new possibilities in terms of quality control and validation.  In this talk, we will present key figures about human and animal vaccines. We will discuss how the industry can take advantage of technologies being supplied to the market, including options for upstream and downstream instrumentation.

Case Studies Illustrating Cell Culture Scale-Up Using Stirred-Tank Single-Use Bioreactors
For cultivation of mammalian cells in biopharmaceutical research and manufacturing, single-use technology possesses several advantages to autoclavable material. Bioreactor scalability is critical to streamlining the adaptation of culture volumes during process development and manufacturing.  We will discuss several scaling approaches, such as constant tip speed, kLa, and power number-based scale-up. Data will be highlighted with two case studies.  First, we determined the power numbers of Eppendorf BioBLU® Single-Use Bioreactors using computational fluid dynamics simulations. Based on these data, we scaled up a process for production of monoclonal antibodies (mAb) in CHO cells from 0.25 L to 3.75 L to 40 L by keeping constant P/V values (impeller power consumption per liquid volume) among the differently sized vessels. Similar cell growth curves and mAb production profiles were achieved at all three scales.  Second, we show experimental data from our German customer CEVEC®.  Using a scale-up strategy based on constant gas flow rates, power input and pH, the process engineers at CEVEC developed a bioprocess for production of lentiviral vectors or recombinant Adeno-Associated Viruses (rAAVs).

High Density Perfusion Cell Culture using Bench-Scale Packed-Bed Single-Use Bioreactors with Methods and Techniques for Scaling the Bench-Scale Process Up to Production-Scale Sterilize-In-Place Bioreactors
Vero cells are anchorage-dependent cells that are widely used as a platform for viral vaccine production.  In stirred-tank bioreactors, they are ordinarily grown on microcarriers.  However, Fibra-Cel® disks are a promising alternative attachment matrix with a high surface-to-volume ratio.  They provide a three-dimensional environment that protects cells from damaging shear forces, helping to achieve high cell densities with both anchorage and non-anchorage dependent cells.  We will discuss cultivating Vero cells in single-use vessels pre-packed with Fibra-Cel using the BioFlo® 320 bioprocess control station with a simple perfusion process and how a cell density of approximately 43 million cells per mL was achieved.  Furthermore, we will examine the constant power per volume (P/V) scale-up of the packed-bed impeller system using the power numbers of the bench-scale single-use bioreactors and production-scale sterilize-in-place (SIP) bioreactors.  Scaling-up cell culture processes using a constant P/V is an industry accepted method which reliably produces comparable yields.  Finally, we will also provide a technique for producing enough cells to inoculate the production-scale SIP packed bed bioreactor using a bench scale system equipped with a spin filter impeller as the cell retention device.  Thus, we were able to demonstrate a perfusion process which used a spin filter impeller to facilitate cell culture production with the packed bed bioreactor at the production-scale.

Small-scale perfusion using an ATF cell retention device with parallel bioreactor system
Experimentation at small scale is crucial for the cost efficient development of bioprocesses, which then can be transferred to larger production volumes. To facilitate perfusion cell culture process development at small scale, we tested the feasibility of connecting an alternating tangential flow (ATF) filtration device with a DASGIP Parallel Bioreactor System. In a working volume of 1 L we reached a peak viable density of 60 x 106 cells/mL. Cell growth, peak density, and antibody production at the 1 L scale were comparable to those in a previously performed perfusion process with a working volume of 3.75 L. Our results demonstrate the feasibility of ATF-based perfusion at small scale using the DASGIP Parallel Bioreactor System.

Advancing viral vector and vaccine process characterization with adherent cells using an integrated perfusion platform - Presented by Corning
With recent progress in cell and gene therapy sector, suitable scalable adherent cell culture platforms must support sufficient characterization of production process from early clinical development through manufacturing. Recent work to integrate Corning’s CellCube® perfusion platform with single use bioreactors and controllers, offers an efficient way to regulate critical factors such as pH and dissolved oxygen. This combined system provide a more robust characterization critical for advance large scale production of viral vector therapies and adjacent vaccine markets.

Simplify and intensify cell culture with XCell™ ATF presented by Repligen®
In use for over 25 years, perfusion is upstream processing which retains cells inside the bioreactor while continually supplying fresh media and removing metabolic byproducts. Because XCell™ ATF technology minimizes cell shear and keeps cells in constant equilibrium with bioreactor contents, faster cell growth and higher densities can be achieved. This results in high cell density perfusion using the same mode of operation as conventional technologies, but with higher productivity.  This presentation will highlight different applications for simplifying and intensifying cell culture operations in both fed-batch and perfusion mode to dramatically increase facility output without direct change to the core production process.

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