Engineering Commons^™

Up And Down Scale Considerations For The Continuous Production Of Glycooptimized Biopharmaceuticals, Steffen Kreye, Rainer Stahn

Integrated Continuous Biomanufacturing III

Product quality especially with respect to glycosylation is a critical product attribute which can have an immense influence on the activity of glycoproteins and should be closely monitored during process development and manufacturing. GlycoExpressTM cells have been developed in this context to provide human host cell lines with robust glycosylation machineries. This work will address challenges in two case studies for up and down scaling of production processes with focus on product quality and continuous processing. In a first section, data of a process transfer from a 200 L stainless steel to a 1000 L single-use bioreactor for a continuous …

Up And Down Scale Considerations For The Continuous Production Of Glycooptimized Biopharmaceuticals, Steffen Kreye, Rainer Stahn

Integrated Continuous Biomanufacturing III

Product quality especially with respect to glycosylation is a critical product attribute which can have an immense influence on the activity of glycoproteins and should be closely monitored during process development and manufacturing. GlycoExpressTM cells have been developed in this context to provide human host cell lines with robust glycosylation machineries. This work will address challenges in two case studies for up and down scaling of production processes with focus on product quality and continuous processing. In a first section, data of a process transfer from a 200 L stainless steel to a 1000 L single-use bioreactor for a continuous …

Moving From A Bioreactor Scale-Up/Scale-Down Approach To A More Holistic Operational Design Space View, Gene Schaefer, Rick Yeager, Ted Deloggio

Cell Culture Engineering XV

Scale-up of bioreactor processes for biologic products has been an area of focus in the industry for over 50 years and the results have generally been successful through the application of a number of specific correlations across scales for mass transfer, mixing, etc. However, the ability to visualize and understand the bioreactor design space in terms of these parameters has generally been limited as has any general understanding of how close a given process comes to certain sensitivity limits at various scales. As part of a project which involved establishing scale-down models at two different sites with long-term objectives of …

Process Optimization For Semi-Continuous Virus Production At High Cell Densities, Daniel Vazquez

Cell Culture Engineering XV

Background. Unlike production of recombinant proteins, continuous production of viral vaccines at high cell densities (HCD) is still constrained by host cell lysis during virus propagation and limited virus recovery from culture broth. Nevertheless, advanced fed-batch [1] and perfusion strategies can be applied to achieve a high-yield virus production processes. In this study, the development of a high-yield semi-continuous process for the production and purification of the modified vaccinia Ankara virus isolate MVA-CR19 and influenza A/PR/8 in HCD cultivations of the suspension cell line AGE1.CR.pIX (ProBioGen AG, Berlin) is presented.

Methods. Depending on the required scale, high cell …

Overcoming Scale-Up Challenges With A Non-Robust Cell Line, Sigma Mostafa, Brian Baker, Abhinav Shukla

Cell Culture Engineering XV

Cell culture scale up success depends on robust cell line, process, and equipment. Mixing characteristics of bench scale through large scale bioreactors need to be well understood and a reliable scale up parameter needs to be used. The cell culture raw material lot to lot variability as well as hydration protocol reliability needs to be considered. The cell culture process parameter ranges need to be broad enough to account for system variability and the process should not run at the edge of failure. In our experience, greater than 95% of cell lines are robust and easily scalable. Cell line robustness …

A Holistic Approach To The Scale-Up Of A Microcarrier-Based Perfusion Cell Culture Process For The Production Of A Therapeutic Enzyme, Jin Yin

Cell Culture Engineering XV

This case study describes our holistic approach of scaling up a microcarrier-based perfusion cell culture process for the production of a therapeutic enzyme directly from a 12L benchtop glass bioreactor to a commercial scale stainless steel bioreactor. Besides conventional scale-up challenges such as mixing, shear stress and mass transfer, the scale-up of this microcarrier-based cell culture process posed its own challenges, e.g. the original design of the commercial scale stainless steel bioreactor had difficulty suspending microcarriers. Additional challenges included achieving effective cell attachment after seeding, balancing microcarrier stratification and culture turbulence, and minimizing bubble shear stress and foaming. To mitigate …

Challenges In The Use Of Scale-Down Models For Understanding And Mitigating Process Variations Of A Monoclonal Antibody Production Process, Peter Russo

Cell Culture Engineering XV

Scale-down models are commonly used to execute process characterization studies, as well as to screen raw materials and to conduct investigations in support of manufacturing operations. During the clinical manufacture of a monoclonal antibody, consistent product quality was maintained; however, large variations in bioreactor harvest titer (> 2X) were observed between lots. Root cause analysis of this variation did not establish a linkage between culture performance and manufacturing execution or deviations. Further, small scale satellite reactor performance displayed lesser variability; thereby eliminating seed train and raw materials as potential root causes. Small scale investigations, using multiple scale-down models, were not …

Scale-Up And Scale-Down Topics Facing The Industry, Markus Michael Muller

Cell Culture Engineering XV

Scale-up and scale-down is bringing several challenges along. We present some recent examples where these challenges occur and how extensive hardware characterization by for example computational fluid dynamics has leads to get a better understanding of the processes themselves.

The increasing implementation of small scale bioreactor systems like ambr15 and ambr250 as well as the usage of new single use bioreactors (>2-L) in parallel to established glass vessels has led to additional challenges in scale-up and scale-down. The mini-bioreactor systems, originally used for screening approaches, are going to be implemented as small scale testing systems but could also be …

Process Optimization For Semi-Continuous Virus Production At High Cell Densities, Daniel Vazquez, Michael Pieler, Yvonne Genzel, Ingo Jordan, Volker Sandig, Udo Reichl

Cell Culture Engineering XV

Background. Unlike production of recombinant proteins, continuous production of viral vaccines at high cell densities (HCD) is still constrained by host cell lysis during virus propagation and limited virus recovery from culture broth. Nevertheless, advanced fed-batch [1] and perfusion strategies can be applied to achieve a high-yield virus production processes. In this study, the development of a high-yield semi-continuous process for the production and purification of the modified vaccinia Ankara virus isolate MVA-CR19 and influenza A/PR/8 in HCD cultivations of the suspension cell line AGE1.CR.pIX (ProBioGen AG, Berlin) is presented.

Methods. Depending on the required scale, high cell …

Field Assisted Sintering: Challenges In Scale–Up From Buttons To Body Armor, Christopher Haines

Electric Field Assisted Sintering and Related Phenomena Far From Equilibrium

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Field assisted sintering technology (FAST) aka spark plasma sintering (SPS) has begun transitioning out of academic research institutions and into industry and government laboratories. With this transition, there has been considerable challenges in scaling up the sample sizes from ‘buttons’ to larger prototype parts. The primary challenges lie in areas such as temperature/density uniformity, tooling/die design, and near-net-shaping. This paper will report on the results of a 5-year manufacturing technology development program which scaled up parts from 30 mm diameter buttons to 250 mm x 200 mm tiles. We will discuss the influence of tooling material and geometry on …

Evaluation Of Single-Use Bioreactors For The Production Of A Hepatitis C Vaccine Candidate, Ricardo Silva, Joao Clemente, Rute Castro, Lenaig Savary, Alex Xenopoulos, Cristina Peixoto, Manuel Carrondo, Paula Alves

Single-Use Technologies: Bridging Polymer Science to Biotechnology Applications

During the last decade, the usage of single-use bioreactors has been increasing in the biopharmaceutical industry. This technology offers some appealing advantages over their conventional counterparts made of glass or stainless steel, such as operational flexibility, faster batch-to-batch turnaround times and the reduction of clean-up and validation characteristic of single-use materials.

The present work examines a stirred single-use bioreactor for its suitability for the production of a Hepatitis C Virus-Like Particles (VLPs) vaccine candidate using the baculovirus expression system with Sf9 cells.

In this sense a 2L glass stirred tank and a Mobius^® 3L Bioreactor are compared in terms …