Application Note

HEK293 Suspension Cell Culture Using The BioFlo® 320 Bioprocess Controller With BioBLU® 3c Single-Use Bioreactors

By Jorge L. Escobar Ivirico and Ma Sha, Eppendorf, Inc.


Developing innovative preventive solutions through new vaccine technologies or gene therapy platforms that respond to new diseases is an important challenge in present day biotechnology.  To produce high-quality biotherapeutics, process development involves several demanding components including cost, cell line development, small scale exploration, effective scaling, and optimization of upstream/downstream processes. The selection of the host cell is a key factor based on its capabilities and properties, including its ability to grow in suspension or adhere to a substrate.

HEK293 is one of the most versatile mammalian cell lines with a wide range of applications including expression of recombinant proteins, antibodies and viruses. However, a major limitation of the cell line is its tendency to clump when converted to suspension format and therefore has been limited to adherent cell culture. To achieve large scale protein production, a new suspension-adapted HEK293 cell line, Expi293F™, was developed by Thermo Fisher. The new HEK293 cell line appears to be a robust suspension cell line capable of achieving greater per cell productivity in high density culture without clumping.

In this study, we evaluated the cell line using bioreactor batch culture. We used a BioFlo ® 320 bioprocess control system and a BioBLU ® 3c Single-Use Bioreactor to carry out Expi293F batch culture in 2 L scale. In addition we monitored and analyzed the metabolites as well as cell density and viability during 11 days of culture. Our goal in this project was to evaluate the suspension culture of Expi293F cell line using Eppendorf bioprocess equipment and assess its suitability in virus production.

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