Fed‐batch bioreactor process scale‐up from 3‐L to 2,500‐L scale for monoclonal antibody production from cell culture

Yang, Jeng‐Dar; Lu, Canghai; Stasny, Brad; Henley, Joseph; Guinto, Woodrow; González, Carlos Regalado; Gleason, Joseph; Fung, M. A.; Collopy, Brett; Benjamino, Michael; Gangi, Jennifer; Hanson, Melissa C.; Ille, Elisabeth

doi:10.1002/bit.21413

Cited by 85 publications

(60 citation statements)

References 18 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…The agitation was scaled-up keeping the tip speed constant. As shown by Yang et al (2007), scaling up agitation based on constant tip speed gives a constant shear rate but leads to longer mixing time at larger scale. This difference in mixing characteristics can possibly explain the reduced growth rates at the 100 L scale.…”

Section: Discussionmentioning

confidence: 99%

Development of a generic transient transfection process at 100 L scale

et al. 2008

View full text Add to dashboard Cite

We have developed a generic transient transfection process at 100 L scale, using HEK293-EBNA cells and PEI as the transfection reagent for the production of recombinant IgG. The process, including large-scale plasmid preparation, expression at bioreactor scale, capture, purification and, if necessary, endotoxin removal allows reproducible production of more than 0.5 g IgG for in vitro and in vivo studies. We compared the performance of two HEK cell lines, investigated the effect of conditioned medium, optimized the DNA:PEI ratio and implemented a feed strategy to prolong the culture time to increase product yield. The transient transfection protocol developed enables a closed process from seeding culture to protein capture. The challenge of performing a medium exchange before transfection at large scale is solved by applying a continuous centrifugation step between the seeding bioreactor and the production bioreactor. After 7-8 days the harvest and capture is performed in a one-step operation using a Streamline expanded bed chromatography system. Following a polishing step the purified antibody is transferred to the final formulation buffer. The method has shown to be reproducible at 10, 50, and 100 L scale expressing between 5 and 8 mg L -1 IgG.

show abstract

Section: Discussionmentioning

confidence: 99%

Development of a generic transient transfection process at 100 L scale

et al. 2008

View full text Add to dashboard Cite

show abstract

“…[4][5][6][7][8] The selection of expression system is determined by its ability to deliver high productivity with acceptable product quality attributes and the preferences of individual companies, which is often influenced by their historical experiences.…”

Section: Mammalian Expression Systemsmentioning

confidence: 99%

Cell Culture Processes in Monoclonal Antibody Production

Li¹,

Shen²,

Amanullah³

2013

Pharmaceutical Sciences Encyclopedia

View full text Add to dashboard Cite

This chapter outlines cell culture processes in monoclonal antibody production. Cell culture process development starts with cell line generation and selection, followed by media and culture condition optimization in small‐scale systems, including 96‐well plate, shaker flasks, and bench‐scale bioreactors, for high throughput screening purposes. Once the process conditions are defined, the process is often transferred to the pilot scale to get scale‐up information and to produce toxicology materials and later to cGMP manufacturing for production of clinical material. As development of a commercial cell culture process for production of a biological product is completed at the laboratory and pilot scales, the commercialization process begins with process characterization, scale‐up, technology transfer, and validation for the manufacturing process. The strong demand of therapeutic antibody production, relatively modest titers, and reduction of cost of goods has resulted in a trend toward large‐scale manufacturing. Today, the largest biotech companies are using several hundred to 2,000 L scale bioreactors for early clinical production and 10,000‐25,000 L stirred tank bioreactors to produce commercial products.

show abstract

“…In addition, mixing time is not a readily measurable variable that is not convenient for scaling (Varley and Birch, 1999). For example, when scaling up a cell culture process from 3-to 2,500-L, constant mixing time resulted in atypical and unpractical high agitation speed (190 rpm) (Yang et al, 2007). It is not possible to maintain all variables constant simultaneously due to restriction in design and configuration of different bioreactors.…”

Section: Introductionmentioning

confidence: 99%

Scale‐up analysis for a CHO cell culture process in large‐scale bioreactors

Xing

Kenty

et al. 2009

Biotech & Bioengineering

184

143

View full text Add to dashboard Cite

Bioprocess scale-up is a fundamental component of process development in the biotechnology industry. When scaling up a mammalian cell culture process, it is important to consider factors such as mixing time, oxygen transfer, and carbon dioxide removal. In this study, cell-free mixing studies were performed in production scale 5,000-L bioreactors to evaluate scale-up issues. Using the current bioreactor configuration, the 5,000-L bioreactor had a lower oxygen transfer coefficient, longer mixing time, and lower carbon dioxide removal rate than that was observed in bench scale 5- and 20-L bioreactors. The oxygen transfer threshold analysis indicates that the current 5,000-L configuration can only support a maximum viable cell density of 7 x 10(6) cells mL(-1). Moreover, experiments using a dual probe technique demonstrated that pH and dissolved oxygen gradients may exist in 5,000-L bioreactors using the current configuration. Empirical equations were developed to predict mixing time, oxygen transfer coefficient, and carbon dioxide removal rate under different mixing-related engineering parameters in the 5,000-L bioreactors. These equations indicate that increasing bottom air sparging rate is more efficient than increasing power input in improving oxygen transfer and carbon dioxide removal. Furthermore, as the liquid volume increases in a production bioreactor operated in fed-batch mode, bulk mixing becomes a challenge. The mixing studies suggest that the engineering parameters related to bulk mixing and carbon dioxide removal in the 5,000-L bioreactors may need optimizing to mitigate the risk of different performance upon process scale-up.

show abstract

Fed‐batch bioreactor process scale‐up from 3‐L to 2,500‐L scale for monoclonal antibody production from cell culture

Cited by 85 publications

References 18 publications

Development of a generic transient transfection process at 100 L scale

Development of a generic transient transfection process at 100 L scale

Cell Culture Processes in Monoclonal Antibody Production

Scale‐up analysis for a CHO cell culture process in large‐scale bioreactors

Contact Info

Product

Resources

About